THEMATIC POSTER SESSIONS TUESDAY
Poster Session 50
Immunogenomics and immunoproteomics
1263
HS1,2 allele *2 is associated with Balkan
endemic nephropathy
Pandolfi, F1; Lolli, S1; Serone, E2; Giambra, V2; Cianci,
R1; Pavlovic, N3; Frezza, D2
1
Catholic University of Rome, Internal Medicine, Rome,
Italy; 2Tor Vergata University, Biology Enrico Calef,
Rome, Italy; 3University of Nis, Institute of Nephrology
and Haemodialysis, Nis, Serbia and Montenegro
Background: Balkan endemic nephropathy
(BEN) is a major cause of chronic renal
failure in Balcanic regions of Europe.
Despite intensive investigation the patho-
genesis remains elusive: toxic components
in the diet as well as immunological factor
leading to hyper-responses to inammation
and autoantibody production have been
reported to play a role together with
genetic factors. 3Regulatory Region of the
Ig heavy chain harbors three enhancers
involved in Ig class switch and B cell matu-
ration. One of these enhancers: HS1.2 is
polymorphic with four alleles showing dif-
ferent afnity to transcription factors.
HS1.2 alleles are involved in the regulation
of Ig maturation and production. Recent
data suggest that variation of allelic fre-
quency is associated with alteration of
immuno-responses. In particular, allele 2
frequency is increased in several immuno-
logical diseases.
Methods: We investigated HS1.2 allelic fre-
quencies by genotyping 95 BEN patients in
dialysis therapy, 107 subjects of a family
affected by BEN and 111 matched con-
trols.
Results: Results show that HS1.2 allele *2
frequency is signicantly increased: 0.54 in
the family; 0.46 in Ben patients in dialysis
versus 0.40 in controls and on the other
hand allele *1 frequency is highly
decreased in patients in dialysis (0.27) and
BEN aficted patients (0.33) when com-
pared to the control (0.47).
Conclusions: Our results show that BEN
can be added to the growing list of
immune diseases as celiac disease, psoriasis,
rheumatoid arthritis, systemic sclerosis
where increased allele *2 is associated with
an abnormally increased immune-response.
482
1264
Prostaglandins receptors polymorphisms
in patients with cutaneous non-steroidal
anti-inflammatory drugs-induced hyper-
sensitivity reactions
Cornejo-Garca, J1; Jagemann, L1; Dona, I2; Blanca-
cies in the SNPs analyzed were similar to
Lopez, N3; Laguna, J4; Fernandez, J5; Rosado, A6;
Terrados, S7; Gandolfo, M8; Blanca, M2; Canto, G9
1
Fundacion IMABIS- Hospital Carlos Haya, Laboratorio
de Investigacion, Malaga, Spain; 2Hospital Carlos Haya,
Servicio de Alergologa, Malaga, Spain; 3Hospital
Infanta Leonor, Servicio de Alergia, Madrid, Spain;
4
Hospital Central Cruz Roja, Servicio de Alergia,
Madrid, Spain; 5Hospital de Elche, Servicio de Alergia,
Alicante, Spain; 6Fundacion Hospital de Alcorcon,
Servicio de Alergia, Madrid, Spain; 7Hospital Ramon y
Cajal, Servicio de Alergia, Madrid, Spain; 8Hospital de
Fuenlabrada, Servicio de Alergia, Madrid, Spain;
9
Hospital Infanta Leonor, Servicio de Alergia, Malaga,
Spain
Background: Non-steroidal anti-inamma-
tory drugs (NSAIDs) are actually the com-
pounds most frequently involved in
hypersensitivity drug reactions, and there
are at least three different mechanisms
involved: IgE- mediated, and T cell-medi-
ated (immunological specic mechanisms)
and cross-intolerance (CI) (immunological
non-specic). In the latter group skin is the
main affected organ. Associations between
CI and different SNPs in arachidonic acid
pathway or prostaglandins receptors
(PGsR)-related genes had been described
in patients with respiratory symptoms or
chronic urticaria, but up today the number
of cases is rather limited and selected pop-
ulations are heterogeneous. In this study,
we analysed the association between CI
and several SNPs in PGsR in a large group
of patients with acute urticaria/angioedem-
a, dened as skin episode in absence of air-
ways involvement or coexistent chronic
urticaria.
Method: The population was obtained
from several Allergy Services integrated
into the Spanish network RIRAAF. Cases
included had to have at least two episodes
of CI with cutaneous symptoms after the
intake of two or more NSAIDs from dif-
ferent chemical groups. We studied several
SNPs in PTGER1, PTGER2, PTGER3,
PTGER4, PTGDR, and PTGFR genes by
using the TaqManOpenArray system.
Allergy 66, Suppl. 94 (2011): 482642  2011 John Wiley & Sons A/S
Result: We included 249 subjects with urti-
caria/angioedema, and 247 age- and sex-
matched healthy controls. We found statis-
tically signicant differences in genotypic
frequencies in PTGDR rs8004654.
Conclusion: Genotypic and allelic frequen-
others studied populations. Although an
association was found between acute urti-
caria/angioedema and the rs8004654 poly-
morphism, further studies are required to
analyze the role of genetic variants in
PGsR and other related genes in this clini-
cal entity.
This study was supported by grants
from the Spanish Health Ministry Fund
for Health in Spain (FIS) network RI-
RAAF (RD07/0064), PI071220 and PD09/
02419.
1265
Lipoxygenase pathway polymorphisms in
hypersensitivity to non-steroidal anti-
inflammatory drugs
Jageman, L1; Cornejo-Garca, J1; Blanca-Lopez, N2;
Dona, I3; Fernandez, J4; Laguna, J5; Rodrguez-Gueant,
R6; Gueant, J6; Canto, G2; Blanca, M3
1
Fundacion IMABIS-Carlos Haya Hospital, Research
Laboratory, Malaga, Spain; 2Hospital Infanta Leonor,
Allergy Service, Madrid, Spain; 3Hospital Carlos Haya,
Allergy Service, Malaga, Spain; 4Hospital de Elche,
Allergy Service, Alicante, Spain; 5Hospital Central Cruz
Roja, Allergy Service, Madrid, Spain; 6University of
Nancy-University Hospital Center of Nancy, Nancy,
France
Background: Non-steroidal anti-inamma-
tory drugs (NSAIDs) are the most fre-
quently involved agents in hypersensitivity
drug reactions. There are at least three
mechanisms: IgE-mediated, T cell-depen-
dent, and cross-intolerance (CI) reactions,
being the latter the most frequent. A num-
ber of studies have described the associa-
tion of different SNPs in genes related with
the arachidonic acid pathway, although the
size of the populations included was lim-
ited and not always homogeneous. In this
work we analyzed the potential association
of several SNPs in the metabolic pathway
of arachidonic acid in a large group of
patients with diagnosed CI to NSAIDs,
Poster Session 50 Immunogenomics and immunoproteomics
and their potential implication in this
pathology.
Method: Patients were recruited in several
Allergy Services that are integrated into
the Spanish network RIRAAF, and diag-
nosed of CI to NSAIDs, with two or more
episodes after the intake of two or more
non-chemically-related NSAIDs. We
selected a group of SNPs in genes related
to lipoxygenase pathway: ALOX5,
ALOX12, and ALOX15. Genotyping was
carried out by means of the Taq-
ManOpenArray system.
Result: We included a total of 324 patients
and 271 age- and sex-matched healthy con-
trols. We found statistically signicant dif-
ferences in genotypic frequencies between
patients and controls in rs4986832 and
rs6413416 (ALOX5), and rs1126667 and
rs3892408 (ALOX12).
Conclusion: Associations between several
of the SNPs studied and CI were found.
Further studies are required to analyze the
role of SNPs in lipoxygenase enzymes in
the etiopathogenesis of hypersensitivity to
NSAIDs. This study was supported by
grants from the Spanish Health Ministry
Fund for Health in Spain (FIS) network
RIRAAF (RD07/0064), PI071220 and
PD09/02419.
1266
Angiolymphoid hyperplasia with eosino-
philia in an Iranian diabetic dentist
Nabavi, M
Tehran Medical University, Pediatric Immunology and
Allergy, Islamic Republic of Iran
A 56-year-old woman presented with mul-
tiple well dened esh like red-purple nod-
ules resembling hemangiomas over her
external ear which are easily hemorrhagic
and intensely pruritic. They started to
develop 2 years ago as a non-tender swell-
ing of left parotid area, soon expanding to
nearby structures mainly the left auricle.
Initially, red to pink maculopapular lesions
appeared over the upper pole of the left
external ear with some pruritus and scanty
hemorrhages. They did progress rapidly,
enumerate and enlarge to become a group
of eshy nodules ranging from 5 to 15 mm
in size. They bled spontaneously nearly
every day and were intensely pruritic.
Anterior cervical lymph node at the same
side involved with no external appearance
except some bulging without any pruritus
and bleeding.
The patient is a dentist with long lasting
type 1 diabetes mellitus, receiving insulin
since more than 40 years ago. Hemoglobin
A1C shows prompt diabetes control. Regu-
lar and NPH insulin has been replaced by
Insulin aspart (marketed by Novo Nordisk
Allergy 66, Suppl. 94 (2011): 482642  2011 John Wiley & Sons A/S
as NovoLog/NovoRapid) and Lentos
Insulin since 2 years ago; that is shortly
before developing her symptoms.
Lab data revealed elevated eosinophils
(610%) and normal serum immunoglobu-
lins including total serum IgE. Erythrocyte
sedimentation rate is highly elevated
(ESR = 88). Hashimoto thyroiditis diag-
nosed recently according to some clinical
and paraclinical ndings such as elevated
anti-TPO and anti-TGB antibodies which
may be due to autoimmune thyroid disor-
der.
There is evidence of impaired creatinine
clearance with 4.8 mg/dl creatinine and
urea concentration of 128 mg/dl which
may be attributed to the late complications
of type 1 diabetes mellitus. She remained
normotensive with no sequelae related to
her possible diabetic nephropathy. Micro-
scopic ndings showed orthokeratosis and
irregular mild epidermal acanthosis. Vascu-
lar proliferation with epithelioid endothe-
lial lining and eosinophil-rich perivascular
inammatory derma reaction was reported
to be consistent with angiolymphoid hyper-
plasia with eosinophils (ALHE).
MRI revealed signal change at left
preauricular area associated with subcuta-
neous soft tissue nodules. Multiple lymph
node involvement was also reported
around the left parotid and cervical chain.
Pulsed dye laser (PDL) and cryotherapy
performed without any benet. Homeo-
pathic management terminated the bleed-
ing tendency.
1267
HLA class I and class II associations in
dengue viral infections in a Sri Lankan
population
Malavige, G1; Rostron, T2; Jayaratne, S3;
Rohanachandra, L3; Fernando, N3; De Silva, A4;
Liyanage, M3; Ogg, G2
1
University of Sri Jayawardanapura and University of
Oxford, Department of Microbiology, Sri Lanka;
2
University of Oxford, United Kingdom; 3University of
Sri Jayawardanapura, Sri Lanka; 4Genetech Research
Institute, Sri Lanka
Background: HLA class I and class II
alleles have been shown to be associated
with the development of dengue hemor-
rhagic fever (DHF)/dengue shock syn-
drome (DSS) in different populations.
However the majority of studies have been
based on limited numbers of patients. In
this study we aimed to investigate the
HLA-class I and class II alleles that are
positively and negatively associated with
the development of DSS in a large cohort
of patients with dengue infection and also
the alleles associated with development of
DHF during primary dengue infections in
a Sri Lankan population.
Methodology: The allele frequencies of
HLA class I and class II alleles were com-
pared in 110 patients with DHF and 119
individuals from the population who had
never reported a symptomatic dengue
infection at the time of recruitment.
Results: We found that HLA-A*31 (cor-
rected P = 0.01) and DRB1*08 (corrected
P = 0.009) were associated with suscepti-
bility to DSS when infected with the den-
gue virus, during secondary dengue
infection. The frequency of DRB1*08 allele
was 28.7 times higher in patients with DSS
than in the normal population. HLA-A*31
allele was increased 16.6-fold in DHF who
developed shock when compared to those
who did not develop shock. A*24 (cor-
rected P = 0.03) and DRB1*12 (corrected
P = 0.041) were strongly associated with
the development of DHF during primary
dengue infection.
Conclusions: These data support a role for
T cells in the pathogenesis of DHF and
DSS. As T cell epitopes to some of these
HLA alleles have already been identied, it
will now be important to investigate how
epitope specic T cells associate with pri-
mary and secondary dengue infections.
1268
Polymorphisms of TAP-1 gene are associ-
ated with mastocytosis. European Com-
petence Network in Mastocytosis
(ECNM) study
Nedoszytko, B1; Niedoszytko, M2; Lange, M1; van
Doormaal, J3; Glen, J1; Zab3otna, M1; Jassem, E2;
Varkonyi, J4; Roszkiewicz, J1; Valent, P5
1
Medical University of Gdansk, Department of
Dermatology, Gdansk, Poland; 2Medical University of
Gdansk, Department of Allergy, Gdansk, Poland;
3
University Medical Center Groningen, Department of
Allergology, Groningen, The Netherlands;
4
Semmelweis University, Department of Internal
Medicine, Budapest, Hungary; 5Medical University of
Vienna, Department of Internal Medicine I, Division of
Hematology & Hemostaseology, Vien, Austria
Background: Mastocytosis (MC) is rare
heterogenous disease associated with
increase of mast cells and their mediators
in different tissues with still unknown path-
ogenesis. The transporters associated with
antigen processing (TAP1 and TAP2) are
proteins that delivers cytosolic peptides to
the lumen of the endoplasmic reticulum
(ER) where there are associated with MHC
class I molecules. It has been suggested
that TAP polymorphisms does affect pep-
tide selection and translocation to ER and
some inammatory diseases are associated
with TAP polymorphisms.
Aim: The aim of the study is to compare
the frequency of TAP1 (333 ile/val, 637
asp/gly) gene polymorphisms in mastocyto-
sis patients and in healthy controls.
Materials and methods: A total 217
patients with mastocytosis and 172 healthy
483

controls were studied. 111 patients were
from Poland, 75 from the Netherlands, 19
from Austria and 12 from Hungary
ECNM centers. Mastocytosis are diag-
nosed on the base of WHO criteria. Study
group included 163 adults and 54 children.
109 patients have systemic (mostly indolent
form) and 96 (included all children) have
cutaneous form of the disease. TAP1 (333
ile/val, 637 asp/gly) gene polymorphisms
were typed using ARMS-PCR method.
Results: The rare 637 gly/gly genotype of
TAP-1 gene was more frequent in the
mastocytosis patients in comparison to the
controls (28% versus 5%, P = 0.0001). No
differences are found between patients with
cutaneous and systemic form of disease.
Conclusion: The results suggest association
of rare 637 gly/gly genotype of TAP-1 gene
with mastocytosis.
1269
Polymorphisms of interleukin 6 gene are
associated with children with mastocyto-
sis. European Competence Network in
Mastocytosis study
Nedoszytko, B1; Niedoszytko, M2; Lange, M1; van
Doormaal, J3; Glen, J1; Zablotna, M1; Jassem, E2;
Varkonyi, J4; Roszkiewicz, J1; Valent, P5
1
Medical University of Gdansk, Department of
Dermatology, Gdansk, Poland; 2Medical University of
Gdansk, Department of Allergy, Gdansk, Poland;
3
University Medical Center Groningen, Department of
Allergology, Groningen, The Netherlands;
4
Semmelweis University, Department of Allergy,
Budapest, Hungary; 5Medical University of Vienna,
5Department of Internal Medicine I, Division of
Hematology & Hemostaseology, Vien, Austria
Background: Mastocytosis (MC) is rare
heterogenous disease associated with
increase of mast cells and their mediators
in different tissues with still unknown path-
ogenesis. Interleukin-6 (IL-6) is multifunc-
tional cytokine that plays roles in the
regulation of hematopoiesis, immune
response, and inammation and is one of
the major mast cell derived mediator which
is elevated in the patients with mastocyto-
sis.
Aim: In our study we compared the fre-
quency of IL-6 )174 G/C promoter gene
polymorphisms in mastocytosis patients
and in healthy controls.
Materials and methods: A total 231
patients with mastocytosis and 161 con-
trols were studied. 125 patients were from
Poland, 75 from the Netherlands, 19 from
Austria and 12 from Hungary European
Competence Network in Mastocytosis
(ECNM) centers. Study group included
177 adults and 54 children. Mastocytosis
are diagnosed on the base of WHO crite-
ria. 130 patients have systemic (mostly
indolent form) and 97 (included all chil-
dren) have cutaneous form of the disease.
The polymorphism )174 G/C in the pro-
484
Poster Session 50 Immunogenomics and immunoproteomics
moter region of TNF-a gene was typed
using ARMS-PCR method.
Results: The high transcription )174 GG
genotype of IL-6 gene was more frequent
in the children with mastocytosis in com-
parison to the controls (27.8% versus
15.5%, P = 0.04). No differences are
observed between other compared groups
(systemic versus cutaneous forms).
Conclusion: The results suggest association
of IL-6 )174 G/C promoter gene polymor-
phisms with pediatric mastocytosis.
1270
Influence of polymorphisms in inflamma-
tory cytokines on risk of duodenal ulcer
among patients with Helicobacter pylori
infections
Yanovich, O1; Titov, L1; Nosova, E1; Doroshko, M1;
DuBuske, L2
1
Research Institute for Epidemiology & Microbiology,
Minsk, Belarus; 2Immunology Research Institute of New
England, Gardner, MA, United States
Background: Helicobacter pylori (H. pylori)
infection is associated with divergent clini-
cal outcomes dependent on the H. pylori
genotype and activity of cytokines in the
gastric mucosa. Genetic polymorphisms of
inammatory cytokines (IL-1B, IL-1RN,
IL-8 and TNF-alpha) also play an impor-
tant role in the host responses to pathogen
invasion, level of cytokine production and
local inammation activity.
Method: Genomic DNA was extracted
from biopsies of 125 patients all having H.
pylori infection associated with various
gastrointestinal diseases. Polymorphisms in
IL-1RN and TNF-alpha genes were ana-
lyzed using the PCR and allele-specic
PCR. The study population included H.
pylori-positive gastritis (n = 88) and duo-
denal ulcer patients (n = 37).
Result: Higher prevalence of the genotype
IL-1RN*2/L was found among patients
with duodenal ulcer in comparison with
chronic gastritis patients (68% versus 47%,
OR = 3.5, CI 1.48.5, P < 0.05).
Homozygote GG in the )308 TNF-alpha
genotype was the most common in both
groups, however there was a signicant dif-
ference among patients with the AA homo-
zygote. The frequency of the AA in )308
TNF-alpha genotype was determined as
0% among patients with chronic gastritis
and 11.1% among patients with duodenal
ulcer (P < 0.05).
Conclusion: This study suggests that cer-
tain allelic forms of the TNF-alpha and
IL-1RN genes may increase an individuals
risk for duodenal ulcer formation during
H. pylori infection. Determination of the
genetic polymorphisms of inammatory
cytokines in patients with H. pylori infec-
Allergy 66, Suppl. 94 (2011): 482642  2011 John Wiley & Sons A/S
tions is a useful means for assessment of
the risk of serious disease progression.
1271
Chediak Higashi Syndrome presenting
with haemophagocytic
lymphohistiocytosis
Aydogmus, C1; Turel, O2; Akcay, A3; Aydogan, G3;
Hatipoglu, N3; Babayigit, A3
1
Bakyrkoy Maternity and Childrens hospital, Pediatric
Infection and Immunology, Istanbul, Turkey; 2Bakyrkoy
Maternity and Childrens hospital, Turkey; 3Turkey
Chediak Higashi Syndrome (CHS) is char-
acterized with hypopigmentation in skin
and hair, recurrent infections and neuro-
logical abnormalities. It is inherited auto-
somal recessively, the mutation in gene
CHS1(LYS) is responsible. Giant granules
are seen in peripheral leukocytes and other
cells. In this report a case with CHS pre-
sented with fever of unknown origin was
presented.
Case report: A six months old male infant
was admitted with fever, cough and
paleness. He was the rst child of consan-
guineous parents. He had outpatient exam-
inations and antibiotic treatment because
of fever during the last 2 months but could
not be cured. Physical examination
revealed pale skin, gray hair and
hepatosplenomegaly. Laboratory tests
resulted pancytopenia, low ESR, increased
PT, PTT, decreased brinogen, hypertrigly-
ceridemia and increased ferritin. Viral
serology was negative, the lymphocyte sub-
groups and immunoglobulin levels were
normal according to his age. Bone marrow
(BM) examination revealed haemophago-
cyting histiocytes. CD25, IL-10, IL-6, IL-8,
TNF alpha levels were higher than normal
in immunological work up. Gray colour of
hair and consanguinity of parents led us to
the diagnosis of CHS or Gricelli syndrome.
Skin biopsy detected melanin loss in hair
follicles and epidermis. Irregular compos-
ites of melanin were seen in the micro-
scopic investigation of hair. The patient
was diagnosed as CHS with the presence
of typical giant granules in neutrophiles
and myeloid cells in peripheral blood
smear and BM examination. HLH 2004
chemotherapy protocol was given. CHS1
mutation analysis was planned but his
family was unwilling for further investiga-
tion since the patient improved after che-
motherapy. CHS was presented since it a a
rare diagnosis. We also like to emphasize
that there are still problems in convincing
families about the severity of diseases of
their children in our country which will
affect the survival.
Poster Session 50 Immunogenomics and immunoproteomics
1272
363C>T polymorphism of TPSAB1 gene
in allergic patients
Hernandez-Hernandez, L1; Garcia-Isidoro, M2; Pascual,
M1; Sanz, C1; Moreno, E1; Davila, I1; Lorente, F1
1
University Hospital of Salamanca, Allergy Department,
Salamanca, Spain; 2University Hospital of Salamanca,
Biochemistry Department, Salamanca, Spain
Background: Tryptase is a serine protease
stored in the secretion granules of mast
cells. The mayor types of tryptase are
beta-tryptase and alpha-tryptase, both
coded by the TPSAB1 gene. Several SNPs
have been described along this gene. The
363C>T SNP is recognized as an EcoRV
RFLP and distinguishes alpha and beta
allele of TPSAB1 gene. The alpha allele
has been previously related to protection
in allergy.
Method: We have studied 125 atopic
patients and 133 non-atopic control indi-
viduals with normal serum tryptase levels.
DNA fragments were amplied by PCR
using primers corresponding to TPSAB1
gene and incubated with the EcoRV
enzyme. Subsequently the fragments were
visualized in an agarose gel. Frequencies
were determined and compared with those
described in the literature.
Result: In the global population, the allelic
frequency of alpha allele was 0.62 while
beta allele frequency was 0.38. The genoti-
pic frequency of beta-beta genotype was
0.76 and 0.24 was to alpha-beta genotype
for the RFLP analysis. There were no sig-
nicant differences in the allelic and geno-
typic distribution between controls and
atopic patients although the alpha allele
presence was slightly higher in the control
group. Statistical signicant association
between SNP 363C>T (RFLP of EcoRV)
and clinic-biological characteristics, IgE
levels, tryptase levels and positive skin tests
to aeroallergens, dander, dust mites, molds
and pollens was not observed. Tryptase
levels in carriers of alpha allele were
4.77 lg/l (2.33 of standard deviation) ver-
sus 6.15 lg/l (5.99 of standard deviation) in
non-carriers individuals (P = 0.057).
Conclusion: Although the alpha allele was
slightly more frequent in our control popu-
lation, we did not nd the previously
describe protective role of this allele in our
population of allergic patients.
1273
Functional analysis of the )95G>T PTGDR
promoter variant in asthmatic patients
Sanz, C1; Garcia Solaesa, V2; Garcia-Isidoro, M2; Benito,
D1; Munoz, F1; Lorente, F1; Davila, I1
1
Complejo Asistencial Universitario de Salamanca,
Servicio de Alergia, Salamanca, Spain; 2Complejo
Asistencial Universitario de Salamanca, Servicio de
Bioquimica, Salamanca, Spain
Allergy 66, Suppl. 94 (2011): 482642  2011 John Wiley & Sons A/S
Background: Allergy and asthma are
thought to be determined by the interac-
tion between several genes and different
environmental factors. Among these genes
PTGDR has been strongly associated with
asthma susceptibility in diverse popula-
tions. In the present study we analyze the
putative association of the recently
described )95G>T PTGDR SNP with
asthma, as well as its implication in puta-
tive transcription binding afnity changes.
Methods: In this study, 602 Caucasian
individuals were included (351 Asthmatic
patients and 251 non-atopic non-asthmatic
controls). Asthma was physician-diagnosed
and allergy was determined by skin prick
testing. )95G>T Genotype frequencies
were determined by PCR and direct
Sequencing. In silico studies considering
the )95G>T variation were performed
using ElDorado and TESS software. Successful isolation of total RNA from
Finally EMSA assays were performed with
nuclear proteins extracts and the )95G>T
surrounded region to detect binding afn-
ity changes.
Results: No signicant associations
between the )95G>T SNP and the asthma
or allergy phenotypes were detected. ElDo-
rado in silico analysis revealed no differ-
ences between the wild type and the
mutant allele at )95G>T, whereas TESS
software predict the binding lost of tran-
scription factors GCF and T-Ag when the
mutant allele is present. However, these
binding afnity differences were not con-
rmed by EMSA assays.
Conclusions: A new )95G>T polymor-
phism has been recently described by our
group in the PTGDR promoter region;
however no genetic association was
detected with asthma or allergy in this
study. Although discrepancies were
detected in the in silico analysis, we did not
detect binding afnity differences in the
EMSA assay what were in agreement with
the ElDorado in silico analysis. This new
SNP seems not to have functional implica-
tions in the PTGDR promoter activity
although discrepancies in the in silico mod-
els needs to be explained.
1274
Optimisation of high-quality nucleic acid
isolation from linden tree (Tilia cordata)
leaves and pollen
Ognjenovic, J; Tantoush, Z; Janko, R; Cirkovic
Velickovic, J; Vukmirica, J
Faculty of Chemistry, Belgrade University,
Biochemistry, Belgrade, Serbia
Background: Tree pollens are among the
most important allergen sources. Linden
trees are widely grown as an ornamental
tree throughout its native range in Europe.
Allergy to linden pollen has been reported,
but little is known about the relevant aller-
gens of this pollen.
Method: Total ribonucleic acid (RNA)
from linden leaves and pollen was isolated
using commercially available plant kit and
guanidinium thiocyanate-phenol-chloro-
form extraction method. Removal of a
genomic DNA contamination was achieved
using enzyme Dnase. Functionality and
high-quality of the isolated RNA from lin-
den pollen and leaves was demonstrated by
rst- and second- strand cDNA synthesis
reactions, as well as RT-PCR.
Result: Conditions for preparative isolation
of high-quality total RNA from European
linden leaves and pollen were determined.
Pure total RNA was isolated from linden
leaves utilizing commercially available plant
mini kit while total RNA isolated from lin-
den pollen using this method was degraded.
both linden pollen and leaves, however,
was achieved following guanidinium thiocy-
anate-phenol-chloroform extraction of total
RNA. Isolated total RNA from both leaves
and pollen was used successfully in rst-
and second- strand cDNA synthesis reac-
tions, as well as, in RT-PCR, demonstrat-
ing that the total RNA isolated using this
method is functional.
Conclusion: Pure and functional total
RNA from Tilia cordata leaves and pollen
(27.8 7.9 lg/g leaves; 25.7 1.1 lg/g
pollen) can be obtained and applicable for
further molecular biology studies.
1275
IgE levels are restricted by a limited
humoral IgE immunological memory
Achatz-Staussberger, G; Feichtner, S; Schwaiger, E;
Wolkerstorfer, S; Vrabel, S; Achatz, G
University of Salzburg, Molecular Biology, Salzburg,
Austria
Background: Serum antibodies, in particu-
lar the isotypes other than IgM, bear the
individual humoral immunological mem-
ory. They are produced by plasma cells,
relatively long-lived cells that integrate the
recent immunological history into a longer-
lasting, protective shield. Unfortunately, in
predisposed individuals, they also perpetu-
ate the production of unwanted, harmful
antibodies, like IgE in allergy and autoan-
tibodies in autoimmune diseases. The
plasma cell, the nal cell type in a long B-
cell differentiation process, can be identi-
ed based on the expression of specic
markers. Several markers that are specic
for the B cell lineage are down regulated
upon plasma cell differentiation. In con-
trast, CD138 is up regulated and serves as
an identifying surface marker for plasma
cells. Although some plasma cells persist in
the spleen, most of them return to their
485

place of birth and home to the bone mar-
row or inamed tissues where they persist
for up to several months in survival niches
as resident, immobile cells.
Method: The functional importance of the
BCR in the generation of humoral immu-
nological memory became apparent in
experiments in which gamma1 and epsilon
cytoplasmic tails were deleted in the mouse
germ line, and a profound deciency in the
development of IgG1- or IgE-antibody
responses was observed. To further investi-
gate the biological function of Ig-tails we
constructed mouse strain KN1, which
expresses a chimeric epsilon-gamma1 B cell
receptor, consisting of the extracellular
domains of the epsilon gene and the trans-
membrane and cytoplasmic domains of the
gamma1 gene.
Result: KN1 mice show an increased serum
IgE level, resulting from an elevated num-
ber of IgE-secreting cells. Whereas the pri-
mary IgE immune response in KN1 mice is
inconspicuous, the secondary response is
far more robust. Most strikingly, IgE-anti-
body secreting cells with gamma1-signalling
history migrate more efciently towards the
chemokine CXCL12, which guides plasma-
blasts to plasma cell niches, than IgE-anti-
body-secreting cells with wild type epsilon
signalling history.
Conclusion: Our observations lead to the
conclusion that IgE plasmablasts have an
intrinsic, lower chance to contribute to the
long-lived plasma cell pool and thus to
humoral immunologic memory than IgG1
plasmablasts. Apparently, an IgE immune
response is in all stages of the response less
efciently regulated, leading to a poor
response.
1276
Investigation of gene expression in HBD-
2 and TLR-2 in the skin of patients with
lichen planus cranes
Dvornikov, A; Kruglova, L; Shachnovich, A; Dontsova, E
Skin and venereal diseases clinic No.1, Department of
Health, Moscow, Russian Federation
Background: In the work carried out to
study the level of expression B-defensins
HBD-2 and TL-2 receptors in the patients
lesions with lichen planus. The study was
conducted by PCR diagnostics.
Method: The most pronounced inhibition
of the expression of B-defensins HBD-2 in
the skin was observed in LRP patients suf-
fering for up to 2 months: 2.091 0.014
(P < 0.01) compared to the normal range
of 3.146 0.023. In the sub-acute stage
(duration of more than 2 months without
a stable remission) the levels of B-defensins
(HBD-2) in the skin did not differ signi-
cantly from the normal values
486
Poster Session 50 Immunogenomics and immunoproteomics
3.223 0.011 (3.146 0.023). In cases of
a chronic LRP during a relapse, the
expression of HBD-2 gene decreased and
was at 2.597 0.012 (P < 0.01) however,
it differed signicantly from the values in
the acute process lasting for up to
2 months 2.091 0.014 (P < 0.01).
Result: With respect to the values of expres-
sion of TL-2 receptors in LPR affected sites
the following trend was observed: regardless
of the stage TL-2 receptors expression
tended to decrease. Meanwhile statistically
signicant differences were observed in the
acute process and long-lasting LRP (a
chronic case in relapse). For example, in the
acute LRP the level of expression of TLR-2
was 2.090 0.011 (P < 0.01) with refer-
ence values of 3.552 0.012, i.e. a 1.7
decrease was observed. In patients with a
chronic LRP the level of expression was not
so much lower (1.3 times), however, it dif-
fered signicantly from the normal values
2.597 0.008 (P < 0.01) versus 3.552
0.012. In sub-acute LRP only a tendency to
inhibition of expression was observed:
TLR-2 3.223 0.010 (P < 0.01). There
were no differences in the level of expression
of B-defensins or TLR-2 receptors in the
skin depending on the LRP clinical form.
Conclusion: Thus we can suppose that in a
LRP relapse, especially during the primary
manifestation of the process, certain sup-
pression against the skins own immune
system is notable. In sub-acute dermatoses,
apparently, immunoregulatory mechanisms
of cellular regulation of inammatory pro-
cesses are restored as a result of activation
of body reserve capacity, which is proven
by improved values of expression levels of
B-defensins and TLR-2 receptors in the
skin.
1277
The potential role of IL-4 and IL-6 gene
polymorphisms in idiopathic pulmonary
fibrosis
Vasakova, M1; Sterclova, M1; Anton, J1; Kolesar, L2;
Slavcev, A2; Skibova, J3; Langova, M4; Striz, I2
1
Thomayer University Hospital, Department of
Respiratory Diseases, Prague, Czech Republic; 2Institute
for Clinical and Experimental Medicine, Department of
clinical and transplantational immunology, Prague,
Czech Republic; 3Institute for Clinical and Experimental
Medicine, Medical Statistic Unit, Prague, Czech
Republic; 4Thomayer University Hospital, Department
of Medical Genetics, Prague, Czech Republic
Background: Idiopathic pulmonary brosis
(IPF) is characterized by pathological pat-
tern of multiple alveolar lesions healing. The
role of regulatory, probrotic and antibrot-
ic cytokines in this process is supposed. Our
aim was to prove a hypothesis about a role
of cytokine gene polymorphisms, namely
that of IL-4, in IPF pathogenesis.
Allergy 66, Suppl. 94 (2011): 482642  2011 John Wiley & Sons A/S
Method: Fifty- six idiopathic pulmonary
brosis (IPF) patients were involved in our
study. All the patients were diagnosed
according to the ATS/ ERS consensus classi-
cation. Genomic DNA was extracted from
peripheral blood using DNA Blood Kit Car-
tridge B350 (Qiagen, Germany) on an auto-
mated instrument BioRobot EZ1. Quality of
DNA samples was checked on spectropho-
tometer and then DNA samples were
archived at 20C. We evaluated polymor-
phisms of 13 different cytokine genes utiliz-
ing CYTOKINE GENOTYPING KIT
(Dynal, Biotech, Norway). The test is
designed as a polymerase chain reaction
(PCR) with sequence- specic primers. In
detail, each well of a 48 well tray contains a
specic primer pair for amplifying the desired
unique sequence. The whole procedure was
performed according to the manufactures
manual. Obtained pattern of positive and
negative PCR is documented and interpreted
according to the manufactures worksheet.
Result: We found a signicantly different
frequency of the investigated IL-4 promo-
tor region polymorphisms ()590) and
()33) and their haplotypes 1 and 2 between
IPF and controls. Moreover, we found also
signicant differences between IPF and
controls for haplotype 1 polymorphisms
frequency at IL-6 ()174) and (nt565).
Conclusion: In our study, we have sup-
ported our previous hypothesis of cytokine
gene polymorphisms pathogenic role in
IPF development. Namely, that the IL-4
promotor polymorphisms can probably be
strongly involved in IPF pathogenesis,
especially in broproliferative healing,
according to their functional relevance,
which results in a shift of immune reac-
tions to so-called TH2 type. The CD4+ T
lymphocytes primed to TH2 cytokine pro-
duction might induce the alternative AMs
activation with consequent probrotic
cytokines and chemokines production. The
role of IL-6 promotor polymorphisms in
IPF pathogenesis is less clear, but the alve-
olar lesions enhancement is possible. This
hypothesis requires further elucidation.
1278
Relation of filaggrin null mutations with
atopy in Croatia
Sabolic Pipinic, I1; Varnai, V1; Turk, R1; Breljak, D2;
Kezic, S3; Macan, J1
1
Institute for Medical Research and Occupational
Health, Occupational Health and Environmental
Medicine, Zagreb, Croatia; 2Institute for Medical
Research and Occupational Health, Molecular
Toxicology Unit, Zagreb, Croatia; 3Coronel Institute of
Occupational Health, Academic Medical Centre,
University of Amsterdam, Amsterdam, The Netherlands
Background: Null mutations in the gene
encoding laggrin (FLG), which result in
the loss of laggrin production and hence
Poster Session 50 Immunogenomics and immunoproteomics
disrupt the epidermal barrier function, have
been strongly associated with atopic derma-
titis and present a predisposing factor in
the development of the atopic march. The
frequencies of the most common null muta-
tions among the Caucasian population in
Western Europe and North America were
analyzed in young adult Croatian popula-
tion, and their relation to skin and respira-
tory atopic diseases was assessed.
Method: FLG null mutations R501X,
2282del4, R2447X and S3247X were geno-
typed in 423 students (305 females and 118
males, median age 19 years) with dened
atopic phenotype (atopic dermatitis, rhini-
tis and asthma) by means of recorded ato-
pic skin and respiratory symptoms with
modied ISAAC questionnaire and posi-
tive skin prick testing (SPT) to one or
more common inhalatory allergens.
Result: We found 11 FLG null mutations
carriers, 1/423 (0.2%) heterozygous for
R501X and 10/423 (2.4%) heterozygous for
2282del4. There were no carriers of
R2447X and S3247X mutations. In total
sample (n = 423), atopic dermatitis was
present in 12%, rhinitis in 17% and asthma
in 7% of subjects. FLG null mutations were
not related to any analyzed atopic pheno-
type. Among 11 FLG null mutations carri-
ers only 3 (27.3%) had atopic dermatitis,
but 9 had eczema/dermatitis symptoms
regardless of positive SPT. Multiple logistic
regression analysis, controlled for gender,
family history of skin allergies and positive
SPT, conrmed FLG null mutations as an
independent risk factor for presence of
eczema/dermatitis symptoms (OR = 22,
95% CI 4.4109.4; P < 0.0001).
Conclusion: The frequency of FLG null
mutation 2282del4 in tested Croatian pop-
ulation was similar to frequency estab-
lished in other European studies on
Caucasian populations (25%). Prevalence
of other tested null mutations was below
1% in our sample, differing from other
studies mainly for R501X. FLG null muta-
tion 2282del4 was strongly associated with
a more frequent expression of eczema/der-
matitis symptoms, but not with any ana-
lyzed atopic phenotype. The majority of
subjects with atopic dermatitis, rhinitis or
asthma were wild type FLG carriers. This
indicates that other genetic and/or environ-
mental factors, rather than the tested FLG
null mutations should be investigated for
contribution to the development of atopic
diseases in Croatian population.
Allergy 66, Suppl. 94 (2011): 482642  2011 John Wiley & Sons A/S
1279
Clinical analysis of a cellular assay in
confirmation of drug allergy
Rudolf, M1; Milavec Puretic, V2; Lipozencic, J2;
Malenica, B1
1
University Hospital Centre Zagreb, Department of
Laboratory Diagnostics, Zagreb, Croatia; 2University
Hospital Centre Zagreb, Department of Dermatology
and Venerology, Zagreb, Croatia
Background: Drug allergies are a group of
symptoms occurs when the immune system
recognize a drug as a foreign substance and
tries to protect organism from it. They
affect 510% of the population. Clinical
symptoms of hypersensitivity reactions to
drugs range from a mild localized rash to
serious effects on vital organs. Drug aller-
gies are fairly uncommon, and identication
of the causative factors is not possible in all
cases. The diagnosis of such hypersensitivity
is based on clinical history, some skin tests,
cellular tests, determination of total and spe-
cic immunoglobulin E (IgE) and other. In
an attempt to nd sensitive, specic and
cost-effective methods to detect hypersensi-
tivity reactions, in vitro tests were developed.
Method: The cellular antigen stimulation
test (CAST) analyzes the mediator released
from effector cells, mainly peripheral blood
cells, when stimulated in vitro with allergens.
The purpose of the present study was to
determine the diagnostic sensitivity of
CAST assay in detection of hypersensitivity
reactions to drugs. Sixty patients at the age
from 4 to 66 years with a history of hyper-
sensitivity reactions to antibiotics (penicillin
V, penicillin G, benzylpenicillin, amoxicillin,
ampicillin, cephalosporin C, cefuroxime,
sulfamethoxazole and trimethoprim) and
analgesic (lys-aspirin, diclofenac and acet-
aminophen) were included in study.
Result: Our results indicate that 73% (16/
22) of patients had positive CAST test to
antibiotics. A positive CAST test to anal-
gesics was found in 36% (4/11) of patients
while 48% (13/27) of patients were positive
to both antibiotics and analgesic.
Conclusion: On the basis of these results
we can see that CAST test used in conr-
mation of the clinical manifestations of
drugs allergy is more sensitive in detection
allergy to antibiotics than to analgesics.
1280
Assessment of modified specific tryptase
assay in diagnosing the etiology of
allergy
Refaat, M; Kamal, A; Fares, M; Attia, M
Ain Shams university, Allergy and Clinical
immunology, Cairo, Egypt
Background: Mast cells are key effectors
cells of the allergic response. When stimu-
lated by specic allergen through the high-
afnity IgE receptors or through other
stimuli, these cells release a number of
potent mediators of inammation.
Amongst these are the serine proteases
tryptase and chymase. Tryptase is the most
abundant mediator stored in mast cells. It
is present as an active enzyme in the mast
cell granules, but displays activity only
extracellularly. Diagnosis of the etiology of
allergy is a vital step in therapy. Numerous
methods are available for diagnosis. Skin
testing has been the primary diagnostic tool
in clinical studies of allergen immunother-
apy. An alternative test is Specic IgE test.
Aim of the work: To assess a new test, the
specic tryptase test, which depends on
libration of tryptase as a result of mast cell
degranulation when it contact with spicic
allergen to which the patient is sensitive to,
it will be carried out as an in vitro test in
order to assess the release of tryptase in the
presence of the allergen. It is a modication
of classic tryptase test which is used in
diagnosing acute allergy e.g. Anaphylaxis
but it is an antigen-specic cellular test.
Methods: A cross sectional study was car-
ried out on 80 persons, 60 patients and 20
controls, their ages were ranged from 9 to
58 years. Patients were classied into three
groups of bronchial asthma, allergic rhini-
tis and chronic urticaria. A skin prick test,
Specic IgE and specic tryptase test was
performed. Data were compared and ana-
lyzed. Results: Accuracy of specic tryp-
tase test as compared by skin prick test is
92.25% while accuracy of specic IgE test
as compared by skin test is 83.08%, in
addition, specic tryptase test detect inha-
ler antigens better than food antigens
(P < 0.001).
Conclusion: This study revealed that the
specic tryptase test is a commercial and
cheap method that sees the cell itself. Its
technical simplicity allows its routine use
by a non-specialized laboratory; also we
can rely on this test in children or in the
patients who can not perform the skin test
in order to diagnose cause of allergy.
487

Allergic inflammation and innate immunity
1281
Increased production of YKL-40 in the
nasal mucosa of patients with allergic
rhinitis
Tyurin, Y1; Kulikov, S1; Sukmanskaya, E1; Reshetnikova,
I2; Fassakhov, R1
1
Russian Federation; 2Kazan Research Institute of
Epidemiology and Microbiology, Allergology, Kazan,
Russian Federation
Background: The chitinase-like protein
YKL-40, also called human cartilage glyco-
protein 39 (HCgp-39) and chitinase 3-like
1 (CHI3L1), which binds chitin but lacks
chitinase activity, has been found to be
either the cause or a biomarker for allergic
response. The aim of our study was to
investigate whether mucosal YKL-40 levels
are increased in patients with allergic rhini-
tis and identify its correlation to amount
of nasal eosinophils and neutrophils.
Method: Fifteen samples of normal nasal
mucosa were obtained from patients who
had normal nasal mucosa and were admit-
ted for augmentation rhinoplasty. Samples
of allergic nasal mucosa were obtained from
twelve patients with perennial allergic rhini-
tis. Measurement of nasal YKL-40 levels
was performed with modication in dupli-
cate using commercially available ELISA
kits for YKL-40 (Quidel, San Diego, CA,
USA). YKL-40 levels were not normally
distributed and the values were compared
among the study groups with the use of the
non-parametric MannWhitney U-test. The
amount of nasal eosinophils and neutroph-
ils and YKL-40 levels were assessed using
Spearmans rank correlation analysis.
Result: There were signicant differences
between the controls and patients with
allergic rhinitis for mucosal YKL-40 levels
and the amount of nasal eosinophil and
neutrophil cells, which are some character-
istics closely associated with allergic
response. The nasal YKL-40 levels in
patients with allergic rhinitis were signi-
cantly (more than 10-fold) higher than in
the controls. In patients with allergic rhini-
tis, the correlation between serum YKL-40
levels and amount of nasal eosinophils and
neutrophils to the predicted value were
investigated. The ndings revealed that
nasal YKL-40 levels correlated with
amount of nasal eosinophils (r = 0.61,
P = 0.065) and with amount of nasal neu-
trophils (r = 0.55, P = 0.085).
488
Poster Session 51
Conclusion: Thus, we conclude that the
level of chitinase-like protein YKL-40 was
upregulated in allergic nasal mucosa com-
pared with normal nasal mucosa, suggest-
ing their roles in the pathogenesis of
allergic rhinitis.
1282
TLR4 and TGFb1 expression varies
between peritoneal macrophages from
different mouse strains in response to
LPS
Soudi, S1; Zavaran Hosseini, A1; Hassan, Z1; Soleimani,
M2; Hashemi, S1
1
Tarbiat Modares University, Faculty of Medical
Sciences, Department of Immunology, Tehran, Islamic
Republic of Iran; 2Tarbiat Modares University,Faculty of
Medical Sciences, Department of Hematology, Tehran,
Islamic Republic of Iran
Background: Macrophages are immune
cells that sense the surrounding microenvi-
ronment through its membrane receptors
and make an effective response to it.TLR4
is one of these receptors that response to
LPS and TGFb1 is one of the macrophage
products in response to environmental
changes. Previous studies showed that
TLR4 activation can affect TGFb1 effec-
tiveness. In this study, we examined the
effect of LPS treatment on TLR4 and
TGFb1 expression level by peritoneal mac-
rophages from different mouse strains.
Method: Six weeks old female BALB/c,
C57BL/6 and DBA mice were purchased
from Pasture Institute of Iran. Six mice
were sacriced from each strain and perito-
neal macrophages were collected in
DMEM containing 10% FBS medium sup-
plemented with NEAA and L-glutamine.
1106 cells /well were plated in12 well
plates. Test groups were treated with
10 lg/ml LPS and controls were not trea-
ted. After LPS treatment, RNA extraction
was performed using QIAzol kit in differ-
ent times and cDNA was prepared. After
cDNA preparation TLR4 and TGFb1
expression were quantied with Real Time
PCR.
Result: Real time analysis showed that
TLR4 expression was signicantly
(P 0.05) increased in all macrophages
after LPS treatment. It was showed that in
test groups, TGF b1 expression was
increased signicantly (P 0.05) in BALB/
c mice as the increase in TLR4 expression.
Allergy 66, Suppl. 94 (2011): 482642  2011 John Wiley & Sons A/S
In contrast, TGF b1 expression was
decreased in test groups of DBA and
C57BL/6 mice.
Conclusion: According to the results, peri-
toneal macrophages response differently to
LPS and show different effect on TGFb1
expression through TLR4 pathway. Since
TGFb1 prepare a suppressive environment,
it seems that macrophages from different
strains have different suppressive/regula-
tory effects on surrounding immune cells.
1283
Correlations between ECP, MPO and
tryptase in nasal and sinus lavages in
allergic and non-allergic patients
Bukovec, Z1; Vagic, D2; Zurak, K2; Pegan, A2; Rasic, I2;
Kalogjera, L2
1
KB S. Milosrdnice, Endo lab, Zagreb, Croatia; 2KB S.
Milosrdnice, ENT, Zagreb, Croatia
Background: The study was designed to
compare cellular markers and correlations
between eosinophils (ECP), mastocytes
(tryptase) and neutrophils (MPO), in sinus
lavage and in nasal lavage. Hypothesis was
that activity of inammatory cells is differ-
ent between asthmatics and non-asthmat-
ics, as well as between nasal and sinus
lavage.
Method: Groups of patients with allergic
(13 patients) and non-allergic rhinitis (7
patients), chronic rhinosinusitis (19
patients) and patients with CRS and
asthma (18 patients) were studied. Patients
with nasal polyposis and receiving systemic
steroid during last weeks were excluded.
The sinus lavage was taken by antral punc-
ture and nasal lavage was taken by nasal
washout, all with 5 cc saline. Samples were
stored at room temperature for 2 h, centri-
fuged at 1000 g for 10 min. and placed in
refrigerator at 20 Tryptase and ECP
were analysed by uoroenzymeimmunoas-
say, and MPO by radioimmunoassay. Sig-
nicance of difference between parameters
tested were calculated with Mann-Whitney
non-parametric test. All conclusions were
based on a signicance level of P < 0.05.
Result: ECP in asthmatics was
64( 72.22) mcg/l, and 48.34 72.2 in
CRS group sinus, while it was 73.77
98.29 in nasal lavage. Tryptase levels in
asthmatics were15.64 30.89 mcg/l, in
CRS 11.07 to 24.58 mcg/l in the sinus,
Poster Session 51 Allergic inammation and innate immunity
and 1.68 3.42 in nasal lavage. MPO was
456 ( 488) mcg/l in the asthmatics,
364( 802) in CRS sinus, and
349.74 484.74 in nasal lavage. ECP and
MPO were signicantly higher in the asth-
matics than in CRS lavage and nasal
lavage, while tryptase is signicantly lower
in nasal lavage. Correlation between ECP
and MPO is highly signicant in the sinus
lavage of asthmatics and non-asthmatics
with CRS (r > 0.85), while it is borderline
signicant in nasal lavage (r = 0.51). Cor-
relations between tryptase and ECP are
signicant only in the sinus lavage of the
asthmatics.
Conclusion: Levels of markers representing
activation of inammatory cells in nasal
and sinus lavages differ between groups
with different pathology. High correlation
of ECP and MPO in sinus lavage, in con-
trast with same correlations in nasal
lavage, indicates difference in the regula-
tion of inammatory process in the sinuses,
compared to nasal cavities.
1284
Correlation between the clinical form of
allergic rhinitis and nasal cytology
1 1 1 1
Gelardi, M ; Petrone, P ; Fiorella, M ; Quaranta, N ;
Russo, C1; Troiano, L1; Ciofalo, A2; Caruso, G1; Salerni,
L1; Tomaiuolo, M3; Maffezzoni, E1; Poletti, D1;
Bocciolini, C1; DallOlio, D1; Landi, M1; Soffia, S4;
Scurati, S4; Puccinelli, P5; Boccardo, R6; Incorvaia, C7;
Frati, F4
1
AICNA, Italy; 2La Sapienza University, Organi di Otorhinolaryngology, Seoul, Republic of Korea
senso Department, Rome, Italy; 3G. Tatarella
Hospital, ORL, Cerignola (FG), Italy; 4Stallergenes,
Medical and Scientific Department, Milan, Italy;
5
Stallergenes, Scientific and Regulatory Department,
Milan, Italy; 6Institute of Pediatrics, Department of
Medical and Surgical Specialty and Public Health,
Perugia, Italy; 7ICP Hospital, Allergy/Pulmonary
Rehabilitation, Milan, Italy
Background: The clinical characteristics
of allergic rhinitis (AR) were classied in
the Allergic Rhinitis and its Impact on
Asthma (ARIA) document according to
duration, that denes the intermittent or
persistent forms, and to the effects of
symptoms on daily activity and sleep, that
dene the mild and moderate/severe forms.
We evaluated the correlation between the
clinical form of AR and the nasal cytology.
Method: Sixty-two patients (34 males, 28
females, mean age 35.2 years) with AR
caused by sensitization to grass pollen were
included in the study. In all patients AR
was dened according to ARIA classica-
tion and nasal cytology was performed by
Rhino-probe sampling, staining and read-
ing by optical microscope. The association
between AR symptoms and cell types in
nasal cytology was analyzed by the chi
squared test.
Result: The distribution of clinical forms
of AR was as follows: 21 patients (33.9%)
Allergy 66, Suppl. 94 (2011): 482642  2011 John Wiley & Sons A/S
had a mild intermittent, 21 (33.9%) a mod-
erate/severe intermittent, 9 (14.5%) a mild
persistent, and 11 (17.7%) a moderate
severe persistent AR. The most common
symptoms were nasal blockage (76.2%),
rhinorrhea (71.4%), sneezing (38.1%), and
nasal itching (9.5%). Nasal cytology
detected neutrophils in 49 patients (29 with
intermittent and 20 with persistent AR),
eosinophils in 41 patients (24 with intermit-
tent and 17 with persistent AR), mast cells
in 21 patients (10 with intermittent and 11
with persistent AR), and lymphocytes/
plasma cells in 26 patients (14 with inter-
mittent and 12 with persistent AR). There
were signicantly more mast cells
(P = 0.014) and lymphocytes/plasma cells
(P = 0.024) in patients with moderate/
severe AR than in patients with mild AR.
Conclusion: These ndings rst demon-
strate that the ARIA classication of AR
severity is associated to different cell
counts in nasal cytology, and particularly
with signicantly higher counts for mast
cells and lymphocyte/plasma cells in mod-
erate/severe AR.
1285
IL-17 deficient mice have lesser airway
remodeling in an experimental allergy
model
Han, D; Hong, S; Lee, J; Kim, D; Lee, C; Min, Y; Rhee, C
Seoul National University Hospital,
University Institute of Experimental Medicine,
Tuberculosis Molecular Epidemiology Laboratory,
Background: It is well known that the air-
way remodeling process develops in asthma
and Th17 cell producing IL-17 have been
found to participate in the development of
allergic asthma. However, the role of IL-17
in the pathogenesis of nasal airway remod-
eling and allergic rhinitis has not yet been
fully elucidated. This study was performed
to investigate the role of IL-17 on the nasal
airway remodeling in an experimental aller-
gic rhinitis model.
Methods: Wild type BALB/c mice (WT-
OVA group) and IL-17 decient mice
(KO-OVA group) were sensitized by intra-
peritoneal ovalbumin (OVA) injection and
subsequently challenged with inhaled OVA
on 7 consecutive days. After then, they
were repeatedly exposed to inhaled OVA
administration 3 days per week for
2 months. Matched control mice (WT-PBS
and KO-PBS groups) were challenged with
phosphate buffered saline instead of OVA.
In the 4 groups (n = 10 per groups), aller-
gic symptoms, and the levels of OVA-spe-
cic immunoglobulin E (IgE) and splenic
cytokines were assessed. Also, airway
remodeling was assessed by expression of
matrix metalloproteinase-9 (MMP-9) and
tissue inhibitor of metalloproteinase-1
(TIMP-1), and subepithelial brosis. In
addition, airway remodeling patterns in the
histology were compared between lung and
nasal mucosa.
Results: IL-17 decient mice showed signif-
icant decrease in allergic symptom scores,
mean serum levels of OVA-specic IgE,
interleukin-4, and subepithelial brosis at
the nasal mucosa. Additionally, airway
remodeling in the nasal tissue was identi-
ed later than in the lung tissue. Immuno-
uorescence staining against MMP-9
revealed stronger intensity in the BALB/c
mice. Furthermore, MMP-9 and TIMP-1
expression at the nasal mucosa was also
down-regulated in the IL-17 decient mice.
Conclusion: Our results, which demonstrate
that IL-17 decient mice have lesser sub-
epithelial brosis, and lower MMP-9 and
TIMP-1 levels at the nasal tissues, suggest
that IL-17 might have a potential role in
the airway remodeling of allergic rhinitis.
1286
Evaluation of neutrophils stimulated by
inactive mycobacterium tuberculosis and
IL-17 in patients with tuberculosis and
cystic fibrosis
Toktay, T1; Bilgic-Gazioglu, S1; Aktas-Cetin, E1;
Kucuksezer, U1; Koser, M1; Yilmaz, A1; Erten, G1;
Koksalan, O2; Kilic, O3; Caglar, E4; Gemicioglu, B5;
Deniz, G1; Camcioglu, Y3
1
Istanbul University Institute of Experimental Medicine,
Immunology Department, Istanbul, Turkey; 2Istanbul
Istanbul, Turkey; 3Istanbul University Cerrahpasa
Medical School, Department of Pediatrics, Division of
Infection, Allergy and Clinical Immunology, Istanbul,
Turkey; 4Yedikule Hospital for Chest Disease and
Thorasic Surgery, Istanbul, Turkey; 5Istanbul University
Cerrahpasa Medical School, Department of Pulmonary
Diseases, Istanbul, Turkey
Background: Tuberculosis has classically
being considered to be Th1 mediated disor-
der. Due to intracellular nature of myco-
bacterial infection little attention has been
paid to the possible role of neutrophils.
Current knowledge regarding neutrophils,
suggests that neutrophils may also contrib-
ute of the pathogenesis of tuberculosis. We
ought to evaluate functions and activation
markers of peripheral neutrophils of
patients with tuberculosis to clarify the role
of neutrophils in the pathogenesis of tuber-
culosis.
Method: Study groups consist of 7 patients
diagnosed as tuberculosis not being under
treatment, 7 cystic brosis patients and 7
healthy control subjects. Neutrophils, iso-
lated from peripheral blood by coll hist-
opaque gradient centrifugation, cultured in
RPMI1640 supplemented by L-glutamine
with inactivated Mycobacterium tuberculo-
sis in the presence and absence of recombi-
nant IL-17. Neutrophil functions such as
489

phagocytosis, chemotaxis and oxidative
burst and also surface expression of the
activation markers including CD11b,
CD63 and CD66b were determined by ow
cytometry.
Result: Oxidative burst results of tubercu-
losis patients neutrophils under bacilli
stimulation is statistically higher than
results obtained after bacilli and IL-17
stimulation (P = 0.043). CD11b mean u-
orescence intensity results after IL-17 stim-
ulation are determined higher than bacilli
and IL-17 stimulation (P = 0.018). When
the results of CD66b mean uorescence
intensity are examined, IL-17 stimulated
neutrophils results are lower than unstimu-
lated neutrophils results (P = 0.018).
Conclusion: Neutrophils functions and acti-
vation markers against IL-17 and bacilli
stimulation from patients with mycobacte-
rial infection were found to be as normal
as healthy donors, but lower levels were
observed in patients with cystic brosis.
Surface CD11b and CD66b expression and
oxidative burst index of neutrophils from
patients with tuberculosis and cystic bro-
sis were inhibited by IL-17.
1287
Allergic inflammation of the airways: is it
one for all?
Koca Kalkan, I; Kalpaklioglu, A
Kirikkale University Hospital, Immunology and Allergic
Diseases Department, Kirikkale, Turkey
Background: Invasive techniques show evi-
dence of a unied allergic airway. Both
allergic and non-allergic rhinitis have been
associated with increased prevalence of
asthma. Nowadays, fractional exhaled
nitric oxide [FeNO] level measured non-
invasively enables to assess airway inam-
mation during an ofce visit. The aim of
this study was to evaluate the bronchial
inammation with FeNO measurement
and factors that inuence it in patients
with different airway conditions, and also
to investigate the impact of atopy, pollen
exposure, and pharmacological treatment
on NO production in lower airways of
patients.
Method: A total of 171 participants were
assessed: 15 healthy volunteers (HV), 95
patients with allergic rhinitis (AR), 18
patients with non-allergic rhinitis (NAR), 27
patients with asthma and AR (A+AR) and
16 patients with asthma and NAR
(A+NAR). After respiratory evaluation,
skin prick tests, spirometry, blood test and
FeNO measurements were performed. Qual-
ity of life (QOL) was evaluated with SF-36,
miniRQOL and miniAQLQ questionnaires.
Result: FeNO levels (ppb) were: 19.4
10.7 (HV), 30.3 23.8 (AR), 14.9 8.2
490
Poster Session 51 Allergic inammation and innate immunity
(NAR), 38.8 31.6 (A+AR), and 40.5
44.1 (A+NAR). A+AR group had signi-
cantly higher FeNO levels than both HV
and NAR groups (P = 0.03 andP < 0.01,
respectively). FENO was signicantly
higher in AR group than NAR group
(P = 0.001). Among A+AR, patients sen-
sitized to perennial allergens had signi-
cantly higher FeNO levels than did
seasonal allergens (48.5 33.9 and
19.5 13.6, P = 0.003). In AR patients
without asthma, no such difference was
found. Atopic patients had signicantly
higher FeNO levels during the allergen sea-
son (AR P = 0.01, A+AR P = 0.05).
Nasal steroids caused signicant reduction
in FeNO levels in only AR patients
(P = 0.09), while no signicance in NAR
and asthma groups. FeNO was correlated
with atopy, male gender, total symptom
score, as well as sneezing and eye com-
plaints. No correlation was found between
FeNO and neither with age, body mass
index, total IgE, number of total sensitized
allergens, FEV1 nor with generic and spe-
cic QoLquestionnaires.
Conclusion: A signicant increased level of
FeNO in allergy season in AR patients
underlies the impact of inammation of
the upper respiratory tract on the lower
respiratory tract. A bidirectional link
between upper and lower airways is con-
rmed by a decrease in FeNO after appli-
cation of nasal corticosteroids. This study
highlighted a link between upper and lower
airways inammation.
1288
Omalizumab and mastocytosis: positive
effects in a patient with a rare sole intes-
tinal manifestation
Angun, O1; van Beek, N1; Zillikens, D1; Grabbe, J2;
Jappe, U1
1
University of Lubeck, Department of Dermatology,
Lubeck, Germany; 2Cantons Hospital Aarau, Clinic for
Dermatology, Aarau, Switzerland
Background: The treatment of symptoms
of intestinal mastocytosis usually consists
of antihistamines, proton pump inhibitors,
antacids, cromoglycate and gluco-corticos-
teroids. Omalizumab is a recombinant
humanized monoclonal antibody that selec-
tively binds soluble serum IgE-antibodies
and is presently approved only for the
treatment of severe asthma. So far, only
very few reports on positive effects in the
treatment of severe systemic mastocytosis
sometimes associated with allergic diseases
exist. We describe a 67-year-old patient
with intestinal mastocytosis and a respec-
tive severe diarrhea, who was previously
unsuccessfully treated with 10 mg of famo-
tidine, 60 mg fexofenadine, 0.5 mg unit-
razepam, 2 mg prednisolone and 900 mg
Allergy 66, Suppl. 94 (2011): 482642  2011 John Wiley & Sons A/S
cromoglycate daily. Interestingly, an acti-
vating KIT D816V point mutation only
occurred in the mast cells of the gastroin-
testinal tract and was not detected in the
bone marrow. As a secondary nding, a
polycythemia vera was diagnosed.
Method: The frequency of diarrhea served
as the efcacy criterion for the treatment
with omalizumab and was initially docu-
mented with an average of ve times in the
morning and three times in the evening.
The patient received 150 mg omalizumab
in 2-week intervals. After the 3rd subcuta-
neous injection the therapy was extended
to every four weeks.
Result: Already after the rst administra-
tion the diarrhea resolved, and after the
second the treatment with prednisolone
could be discontinued.
Conclusion: The mechanism of action of
omalizumab in the treatment of mastocyto-
sis still remains unclear. Downregulation
of the FcEpsilon-receptor could play a role
but also a mast cell stabilizing effect is dis-
cussed. To the best of our knowledge this
is the rst case of a patient with isolated
gastrointestinal mastocytosis, who was suc-
cessfully treated with omalizumab. This
case report together with four previously
published cases conrms that omalizumab
constitutes a well tolerated alternative ther-
apy in treatment-resistant systemic masto-
cytosis.
1289
Bone involvement in patients with clonal
mast cell disorders
Rossini, M1; Bonadonna, P2; Artuso, A3; Caruso, B4;
Schena, D5; Vecchiato, D6; Bonifacio, M7; Gatti, D1;
Senna, G2; Riccio, A6; Pizzolo, G8; Adami, S9;
Passalacqua, G6; Zanotti, R8
1
University of Verona, Dept of Medicine, Rheumatology
Section, Verona, Italy; 2Azienda Osp Universitaria
Integrata of Verona, Allergy Unit, Verona, Italy;
3
University of Verona, Dept of Medicine, Hematology
Section, Verona, Italy; 4Azienda Osp Universitaria
Integrata of Verona, Clinical Chemistry and
Haematology, Verona, Italy; 5University of Verona, Dept
of Medicine, Dermatology Section, Genoa, Italy;
6
Genoa University, Dept of Internal Medicine Allergy &
Respiratory Diseases, Genoa, Italy; 7University of
Verona, Dept of Medicine, Hematology Section, Genoa,
Italy; 8University of Verona, Dept of Medicine,
Hematology Section, Italy; 9University of Verona, Dept
of Medicine, Rheumatology Section, Italy
Background: Several reports suggested a
relatively high occurrence of bone altera-
tions in patients with mastocytosis. We sys-
tematically assessed bone mineral density,
bone turnover markers, and fractures in a
large cohort of patients with clonal mast
cell disorders.
Methods: Eighty-three patients (36 women,
mean age 48 years), 74 with indolent Sys-
temic Mastocytosis and 9 with Monoclonal
Mast Cell Activation Syndrome, were stud-
ied. Bone density was measured by dual
Poster Session 51 Allergic inammation and innate immunity
X-ray absorptiometry at the lumbar spine
and proximal hip. The results were
expressed Z-score (standard deviation
below the age- and gender-matched refer-
ence value from the NHANES Study).
Patients were classied as Mastocytosis-
related osteoporosis if their Z-score at
either the spine or the total hip was <-2
and/or if they had a history of vertebral
fragility fracture.
Results: Three patients were excluded for
concomitant diseases associated to osteo-
porosis. Mastocytosis-related osteoporo-
sis was found in 22/80 (27%) patients
(21% in women, 32% in men). The bone
mineral density was generally lower at the
spine than at the hip, this suggesting a pre-
valent involvement of trabecular bone. One
or more severe vertebral fractures were
found in 11 patients. Two patients had
radiographic and densitometric osteosclero-
sis-like characteristics, and both had higher
serum tryptase and bone turnover levels.
Conclusion: Osteoporosis is frequent in
patients with Clonal Mast Cell Disorders,
particularly at the vertebral site and in
males. Osteosclerosis-like features can be
observed more rarely. Our results suggest
that a densitometric examination is war-
ranted in all patients with mastocytosis,
and evaluated according to Z-scores.
1290
Circulating subsets of monocytes in kid-
ney allograft transplantation
Striz, I1; Sekerkova, A1; Brabcova, E1; Kolesar, L1;
Slavcev, A1; Viklicky, O2
1
Institute for Clinical and Experimental Medicine,
Department of Clinical and Transplant Immunology,
Prague, Czech Republic; 2Institute for Clinical and
Experimental Medicine, Department of Nephrology,
Prague, Czech Republic
Background: Blood monocytes are pheno-
typically heterogeneous population of cells
subdivided according to the expression
level of membrane antigens. In addition to
traditional monocytes characterized by
high expression of CD14 and the absence
of CD16, there is a subset of monocytes
with lower CD14 expression and detectable
membrane expression of CD16. These
CD14+CD16+ monocytes represent
about 15% of peripheral blood monocytes
and their numbers may be increased in dif-
ferent pathological settings. According to
their functional properties, the
CD14+CD16+ cells are activated mono-
cytes. Another monocyte subset is charac-
terized by the expression of CD163 and
represents monocytes with immunosuppres-
sive properties. These cells are in vitro
inducible by corticosteroids and are char-
acterized by an enhanced production of
cytokine IL-10. The aim of our study was
Allergy 66, Suppl. 94 (2011): 482642  2011 John Wiley & Sons A/S
to evaluate the distribution of monocyte
subsets in kidney allograft recipients in
early phases after the organ transplanta-
tion.
Method: Membrane antigens expressed by
monocytes were evaluated by ow cytome-
try in dened time points. The results of
immunophenotyping were correlated with
an immunosuppressive protocol used.
Result: The proportions of peripheral
CD14+CD16+ monocytes were found to
be downregulated during the rst week
after the kidney transplantation. In
patients treated with anti-CD25 antibody
basiliximab, the decrease in percentage of
CD14+CD16+ monocytes was delayed
for one week. In most patients, the tran-
sient downregulation of CD14+CD16+
subset was adjusted to basal values in two
months, only in subjects treated with rab-
bit antithymocyte globuline, the numbers
of CD14+CD16+ monocytes remained
low after 100 days. The proportions of
immunosuppressive
CD14+CD163+monocytes were tran-
siently upregulated early after the kidney
transplantation and remained higher dur-
ing the rst month in most patients. We
have not found any dynamic changes in
proportions of other monocyte subsets,
such as CD14+CD74+ or CD14+
CD36+ described by previous studies.
Conclusion: We assume from our data that
kidney allograft transplantations are asso-
ciated with dynamic regulation of circulat-
ing monocyte subsets CD14+CD16+ and
CD14+CD163+ and these changes are
partially inuenced by an immunosuppres-
sive regime used.
Supported by IGA grant NS/10524-3.
1291
Autoinflammatory diseases: case report
and a review of the literature
Turuntas, V
Clinical center East Sarajevo Foca, Pediatrics
(Allergology), Foca, Bosnia and Herzegovina
The objective was to present a case of peri-
odic fever with aphthous stomatitis, phar-
yngitis and cervical adenitis (PFAPA),
summarize the medical literature on PFAP-
A, review the differential diagnosis and
suggest a diagnostic approach to periodic
fevers in children. Although still without a
known genetic mutation PFAPA, as a part
of periodic fever syndromes, is closely
related to autoinammatory diseases. Chil-
dren with periodic fever or autoinamma-
tory syndromes sometimes present to an
Allergy/Immunology clinic for immuno-
logic evaluation. It is important for the
Allergy/Immunology specialist to be famil-
iar with the clinical presentation, diagnos-
tic approach and treatment of these
conditions. We also conducted a literature
review to investigate the recent advances in
genetics and molecular biology of the
hereditary autoinammatory diseases as a
hereditary inammatory diseases character-
ized by seemingly unprovoked recurrent
episodes of systemic inammation in the
absence of pathogens, autoantibodies or
antigen specic T cells, that arise from
mutations of genes regulating the innate
immune system. The discovery of the
genetic causes of a rare group of immune-
mediated inammatory conditions that
mimic infections and allergic conditions in
their clinical presentation and the molecu-
lar understanding of the function of the
mutated molecules in these diseases has led
to a revolution in our understanding of the
pathogenesis of systemic and local inam-
mation. The proteins mutated in a number
of these so-called autoinammatory dis-
eases are part of, or regulate the activity
of, intracellular molecular complexes, the
inammasomes, that sense danger to the
body and coordinate an initial immune
response.
1292
Congenital immunological reactivity of
newborns of unsuccessful region
Dolynnaya, V; Tuleutaeva, G
State Medical University of Semey, innate immunity
and complement, Semey, Kazakhstan
Statistics of morbidity among newborns of
Semey city and other locality situated near
with former atomic test-site remain unsatis-
factory during many years. Perinatal
pathology is in the structure of causes of
infants mortality more than 50%. Purpose
of work was the assessment of newborns
immunity in comparison with the analo-
gous dates of different regions (Moscow,
Rostov, Perm, Kishinev etc.). We exam-
ined 53 practically healthy babies which
were born by mothers with satisfactory
anamnesis.
As the result of the research of immuno-
competent cells determined by methods of
monoclonal antibodies was found that rel-
ative maintenance of CD3 in patients is
42.6%, it is on 720% lower than in babies
having born in other places, quantity of
CD19 (29.6%) on 1221% and CD8 on
116% is higher. Particular differences of
CD4 quantity were not found.
Individual indexes of immunoglobulins
and circulating immunological complexes
(CIC) in blood of several children varied in
wide boundaries. In comparative aspect of
concentrations of IgG (103.6 1.45 IU/
ml) and IgM (11.08 0.8 IU/ml) so as
491

average level of CIC (20.9 2.5 u) were
higher, that can testify about intrauterine
antigens irritation. IgA (0.68 0.5 IU/ml)
was dened only in single cases.
Thereby, obtained results testify about
disharmony of nuclear-humanitarian coop-
eration of newborns of this area, what in
the given degree connected with relative
high antigenic load. Concluded peculiari-
ties of immunological status show effect on
health status, morbidity and in total statis-
tics of childrens death.
1293
Anti- and pro-apoptotic markers in post-
traumatic stress disorder
Lilit, H1; Gohar, M1; Anna, B1; Margarita, T2; Arthur, K2;
Samvel, S2
1
Institute of Molecular Biology of the National
Academy of Sciences, Armenia; 2Stress Center of the
Ministry of Labour and Social Affairs, Armenia
Background: Post-traumatic stress disorder
(PTSD) is a complex disease in which the
affected persons memory, emotional
responses, intellectual processes, and ner-
vous system have all been disrupted by one
or more traumatic experiences. The molec-
ular mechanisms underlying the pathology
of PTSD are poorly understood. Some
experiments indicate that PTSD can dam-
age the hippocampus and introduce atro-
phy and apoptosis, which is regulated by
different classes of proteins including pro-
apoptotic protein Bcl-2 associated X pro-
tein (Bax) and anti-apoptotic Bcl-2. It has
been shown, that Bcl-2 can block cell death
induced by a variety of stimuli. However, a
Bcl-2-related protein, Bax, forms heterodi-
mers with Bcl-2 and when overexpressed
counteracts the protective effects of Bcl-2.
The aim of this study was to detect the lev-
els of these proteins in PTSD-affected sub-
jects.
Method: Examined patients were combat
veterans, diagnosed according to CAPS.
Age- and gender- matched healthy subjects
without any history of physical or sexual
abuse or other major trauma, dened as
being free of current or past psychiatric
disorders served as a control group. The
levels of Bax and Bcl-2 were determined in
the blood serum of patients and controls.
Result: The results showed that the mean
values of the level of Bax in the blood
serum of patients with PTSD is 1.5
(P = 0.0074) times higher, than in the
healthy subjects group. No signicant dif-
ference in Bcl-2 level between patients and
controls is detected (P = 0.6).
Conclusion: Our data indicate upregulated
apoptosis in chronic PTSD patients.
492
Poster Session 51 Allergic inammation and innate immunity
1294
Human semen inflammation: effects of
proinflammatory cytokines on sperm
membrane integrity, apoptosis and chro-
matin quality
Fraczek, M; Czernikiewicz, A; Kurpisz, M
Institute of Human Genetics Pol. Acad. Sci., Dept. of
Reproductive Biology and Stem Cells, Poznan, Poland
Background: Proinammatory cytokines
are the natural components of seminal
plasma. They were found to regulate a
physiological function of the male gonad
and to act as testicular immunomodulatory
elements. On the other hand, the same
cytokines have been linked with a decrease
of semen quality in infertile men, particu-
larly with urogenital tract infections. As
clinical signicance of proinammatory
cytokines and their inuence on male fertil-
ity is still poorly documented, we have
examined an in vitro effect of six proin-
ammatory cytokines, (IL)-1b, IL-6, IL-8,
IL-12, IL-18 and (TNF)-a on sperm mem-
brane stability, phosphatidylserine (PS)
externalization, DNA fragmentation and
chromatin quality, using a ow cytometry.
Methods: Fresh semen samples were
obtained from seven healthy volunteers
with no sperm abnormalities and any sign
of infection/inammation. Washed sperma-
tozoa were exposed to different cytokines,
at physiological and infection/inammation
concentrations and analyzed after 1, 3 and
24 h incubation. The LIVE/DEAD Sperm
Viability Kit (SYBR14 and propidium
iodide PI) and the merocyanine 540
(M540) test were used to examine sperm
plasma membrane integrity. Sperm apopto-
sis was evaluated with the use of Annexin
V/PI assay and TUNEL-assay. The sperm
chromatin structure assay (SCSA) and
chromomycin A3 staining were used to
analyze the sperm chromatin damage.
Results: There were no statistical differ-
ences in the percentage of live (SYBR-14
positive) or dead (PI-positive) spermatozoa
incubated with particular cytokines versus
sperm incubated alone. Among all the
cytokines tested, only IL-1b and IL-18
affected sperm plasma membrane architec-
ture measured by M540 test even in physi-
ological concentrations, although no
statistical difference was observed. All the
proinammatory cytokines applied in the
study at physiological concentrations had
no inuence on sperm PS externalization.
However, infection-inammation concen-
trations of three cytokines (IL-1b, TNF-a
and IL-18) increased the number of Ann-
exin V-positive/PI-negative cells after 24 h
incubation when compared to the results
observed after 1 h incubation (P < 0.05).
The exposure of spermatozoa to these
cytokines also resulted in the intensication
of DNA fragmentation.
Allergy 66, Suppl. 94 (2011): 482642  2011 John Wiley & Sons A/S
Conclusion: Induction of apoptosis is the
main mechanism by which some proinam-
matory cytokines affect ejaculated sperma-
tozoa especially during the male
reproductive tract infection/inammation.
1295
Reversing persistent debility after past
heavy environmental mold exposure and
immune weakness
Grant, I
New York Medical College, Department of Family and
Community Medicine, Tarrytown, United States
Background: Many mold-exposed patients
have persistent, poorly explained debility
and repeatedly seek care. Molds stimulate
innate immunity independent of allergy,
granulomatous or invasive infection, elabo-
rating myriad toxic products, triggering
severe inammation. Mold particulates,
volatile organic compounds (VOCs) and
toxins cause immunosuppressive destruc-
tive changes in animal models.
Objective: To (1) determine frequency and
risks for persistent debility after heavy
mold exposure and (2) empiric antifungal
therapy efcacy.
Method: Monitored 66 patients (27 M, 39
F, 375 years) with prior heavy old expo-
sure by history, or environmental mold
testing, urine mycotoxin assay and
response to antifungal therapy (nasal
amphotericin B +/) oral azoles).
Result: Of 66 exposures (58 residential, 8
occupational), 24 had intense exposure by
history and 42 by environmental mold test-
ing (42 Aspergillus/Penicillium, 26 Stachy-
botrys, 12 Cladosporium, and 3
Alternaria). Debilitating symptoms (sxs)
persisted in 45/66 (60%) (Descending: Ear-
Nose-Throat (ENT), Pulmonary, GI, Neu-
rological (CNS), Rheumatologic [especially
chronic fatigue syndrome (CFS), brom-
yalgia]). Sxs progressed in 38%, with per-
manent sequelae in 60% with prolonged
heavy exposures or who refused treatment.
Impaired cell-mediated immunity (CMI)
risks were found in 30/66 (45%): protein
deciency (impaired neutrophil response),
vitamin D deciency (impaired innate
immune response), zinc deciency
(impaired neutrophil response); neutrope-
nia; diabetes; steroids. Prolonged debility
was associated with impaired CMI or
intense exposure. Urine mycotoxin excre-
tion in 14/16 (88%) was positive: (7 Aa-
toxin [carcinogen], 14 Ochratoxin
[carcinogen], 7 Tricothecenes [neurotoxin]).
Five severely ill excreted multiple mycotox-
ins. Response to antifungal therapy was
striking: 26/28 (93%) reversed CFS, -
bromyalgia, ENT/Pulm, CNS sxs, menor-
rhagia, rigors, wasting, vertigo. Most
Poster Session 51 Allergic inammation and innate immunity
without impaired CMI improved; some
required 13 months antifungal therapy.
Many with impaired CMI responded, but
relapsed off therapy.
Conclusion: Persistent debility long after
mold exposure is associated with impaired
cell-mediated or innate immunity. Empiric
antifungal treatment dramatically reversed
many persistent, poorly-explained progres-
sive symptoms (e.g. CFS, bromyalgia).
Diagnostic methods to detect mold
byproducts (VOCs, mycotoxins, particu-
lates) and controlled clinical trials evaluat-
ing antifungal therapy are needed.
1296
Study of cytokine status and substance P
in frequently ailing children with acute
respiratory viral infections
Karimova, M
Azerbaijan Medical University, Department of children
diseases 2nd, Baku, Azerbaijan
Background: According to the scientic
works, an acute respiratory viral infection
(ARVI) takes important part in the prog-
nosis of respiratory diseases in childhood.
These infections may be the cause of recur-
rence, complications, changing to the
chronical type of respiratory diseases in
children with weak immune system.
According to the information of the mod-
ern literature, the immune system of the
body acts not solely, but jointly with the
other systems, for example neurologic sys-
tem. The aim of our work was to study the
cytokine status and substance P, mutual
connection they in frequently ailing chil-
dren (FAC) with ARVI and to detect the
inuence of their changes to the respiratory
diseases.
Method: The study included 96 FAC with
respiratory diseases aged 315 years. The
group with seldom ailing children SAC
(the children felt sick of ARVI to three
times in a year) included 30 children. The
control group included 30 healthy children.
All patients have been examined in an
acute period of the disease and during clin-
ical remission. In all patients and healthy
children we have dened the levels of cyto-
kines IL-1b, IL-2, IL-6, IL-8, TNF-a,IFN-
c serum and substance P .
Result: In FAC with respiratory diseases
in the acute period of the disease increase
of levels proinammatory cytokines IL-1b,
IL-6, IL-8, TNF-a and substance P,
decrease of levels IL-2 and IFN-c was
marked. Clinical remission in these chil-
dren is not accompanied by normalization
of cytokine status and substance P. We
also studied the mutual connection
between the cytokines and substance P in
our patients. High positive correlation
Allergy 66, Suppl. 94 (2011): 482642  2011 John Wiley & Sons A/S
between IL-1b and substance P, negative
correlation relations between IL-2 and
IFN-c with substance P is found.
Conclusion: In conclusion, the high levels
of proinammatory cytokines and sub-
stance P in clinical remission of disease tes-
ties to proceeding of inammatory
process that is possible connected with
persistantion of the infections agent. Acute
decrease in level of cytokines IL-2 and
IFN-c in these children is caused by the
presence of a immunodeciency of cellular
type. In this connection it is necessary to
carry out an adequate therapy of FAC
with ARVI.
1297
Conditional pathogenic fungi allergens
as mitogens of cellular link immunity
Bogomolov, A
Vinnytsya National Medical University, Phthisiatry with
Clin. Immunology and Allergy Course, Vinnytsya,
Ukraine
Background: In the last time a large value
gets to the role of immune violations of
system and local character in pathogenesis
of inammatory processes of various etiol-
ogies, which quite often is reason of chron-
isation of processes and development of
resistance. In principle, today there are no
problems with determination of quantita-
tive and high-quality parameters of cellular
link of the immune system. A problem
exists in the real possibility of introduction
of these high-cost immunological methods
in practice of general practitioners work.
In this connection our method of estima-
tion of cellular link state by the skin test-
ing with mitogens can be useful. In quality
of mitogens, taking into account to the fea-
ture of immune answer, their prevalence,
we took the allergens of conditionally-
pathogenic fungi.
Method: Research was conducted on mate-
rial of 120 patients with the different forms
of pulmonary tuberculosis, which passed in
the moment of inspection stationary treat-
ment in Vinnytsya regional antituberculosis
hospital (investigation group) and 40 adult
healthy volunteers (control group). The
rst stage of inspection included the excep-
tion of allergic anamnesis, reception of
hormonal preparations and heavy concom-
itant pathology, the second stage intra-
skin testing with allergens of conditionally-
pathogenic fungi Alternaria, Aspergillus
mixt, Cladosporium, Chrisonilla, Monilia,
Penicillum and Botrytis cinerea during not
more than 10 days after entering of patient
in department. The third stage was labora-
tory testing including immunologic assay.
Result: Immediate type reactions was rare
in all groups. Quantity of delayed - type
hypersensitivity reactions to one patient
was above 2.95 0.90 reactions before
and 4.5 1.1 after 90 days treatment (pri-
mary tuberculosis), 2.65 0.53 reactions
before and 3.87 0.62 after 90-days treat-
ment (relapse tuberculosis), 0.60 0.33
and 1.1 062 accordingly (chronic tuber-
culosis). Testing results with allergens cor-
relate with data of intoxication and
bronchopulmonary syndromes (value of
Spearman coefcient from 0.55 to 0.63),
data of complete blood count (0.50.65),
and indexes of leucocyte migration with
the same allergens (0.50.62).
Conclusion: (1) Mycotic allergens can be
applied as mitogens of T-link immunity.
(2) Most informing as markers are aller-
gens Alternaria sp., Penicillium sp., Chry-
sonilia sitophila, Monililia sitophila. A
complex estimation as mitogens of all
applied allergens is most informing.
1298
Alpha-1-antitrypsin variants and rhinosi-
nusitis
Palma-Carlos, A; Palma-Carlos, L
Clinical Allergy Immunology Center, CAIC, Allergology,
Lisbon, Portugal
Background: Alfa-1-antitrypsin, proteinase
inhibitor alfa-1 (A1PI) plays a central role
in airways protection. Genetic deciency of
A1PI is linked to COPD. The relation
between A1P1 deciency and rhinosinusitis
is not established. A1P1 deciency and
variants have been systematically searched
in out-patients to identify associated dis-
eases.
Method: 58 patients have been studied.
Assay of A1P1 by nephelometry have been
done in all patients. Increase of CRP has
been used as a marker of acute phase reac-
tions masquerading A1PI deciency. A cut
off of 0.2 mg/dl has been considered for
CRP. When A1PI concentration less than
90 mg/dl in absence of inammatory reac-
tions, a genetic study was done. Genomic
DNA was extracted and the research of
mutations Glu 264-Val (S allele) or Glu
342-Lys (Z allele) studied by enzymatic
amplication of DNA by PCR with spe-
cic oligonucleotides and analysis of melt-
ing curves of amplication products. For 2
or more A1PI values less than 90 mg/dl
even in normal PIMM genotype the pres-
ence of a Qo non-secreting null gene as
been considered.
Result: In 51 patients (26M 25F) aged 10
60 years, the presence of abnormal variants
(S, Z or Qo) of A1PI was conrmed. S
allele was present 40 times Z, 20 and Qo 7.
Genotypes: SS-7 MS-18, MZ-10, ZZ-1, SZ-
8, MQo-7 Clinical presentation for these
patients: Chronic rhinosinusitis 29, rhinosi-
493
 Poster Session 51 Allergic inammation and innate immunity

nusitis asthma 14, rhinosinusitis COPD 1,
rhinosinusitis urticaria 1, Asthma 1, COPD
1, Chronic urticaria 4. Rhinosinusitis was
present in 45 patients (88.2%) asthma in
15 (29.4%) urticaria in 5 (9.8%) COPD in
2 (3.9%). The incidence of rhinosinusitis
was independent of genotype: MS 18/18,
MZ 10/10, SS 7/6 MQo 6/7, SZ 5/8 ZZ 0.
Discussion: A1PI deciency due to genetic
variants is usually related to COPD,
asthma and liver diseases. Present study
suggests that A1PI deciency could also be
associated with chronic rhinosinusitis. The
inammatory process triggered by neutro-
phil protease and mast cell chymase is
incompletely blocked by decient A1PI
also in nose and sinus. Larger series can
conrm this hypothesis.

494 Allergy 66, Suppl. 94 (2011): 482642  2011 John Wiley & Sons A/S

Mast cells and eosinophils
1299
Eosinophil extracellular DNA traps in skin
diseases
Simon, D1; Hosli, S2; Yousefi, S2; Simon, H2
1
University of Bern, Dept. of Dermatology, Inselspital,
Bern University Hospital, Bern, Switzerland; 2University
of Bern, Institute of Pharmacology, Bern, Switzerland
Background: In the skin, eosinophils are
found in a broad spectrum of diseases,
including infectious diseases. In this study,
we investigated whether eosinophil extra-
cellular traps (EETs), structures containing
DNA in association with eosinophil gran-
ule proteins able to bind and kill bacteria
are present in the skin under various path-
ologic conditions.
Method: Immunouorescence staining was
performed on sections of paraformalde-
hyde-xed and parafne-embedded skin
biopsy tissues of 25 different eosinophilic
skin diseases using propidium iodide (PI)
and an antibody to eosinophil cationic pro-
tein (ECP). Slides were evaluated by laser
scanning microscopy.
Result: Eosinophils releasing DNA together
with ECP were detected in infectious skin dis-
eases, such as ectoparasitosis and larva mig-
rans. Further, we observed the extracellular
DNA structures in allergic/reactive diseases
(Wells syndrome, hypereosinophilic syn-
drome, positive reaction of atopy patch test,
allergic contact dermatitis, drug hypersensi-
tivity), and in autoimmune diseases (bullous
pemphigoid, pemphigus foliaceus, dermatitis
herpetiformis). The average number of eosin-
ophils releasing DNA in the skin was usually
below 10%, only in Wells syndrome the pro-
portion was up to 30%. In areas with clusters
of eosinophils, up to 50% of the eosinophils
were seen to generate EETs.
Conclusion: EETs are seen in both infec-
tious and non-infectious inammatory skin
diseases, and particularly common in
Wells syndrome.
1300
IL-31 induces chemotaxis and superoxide
anion release of human eosinophils
Kunsleben, N; Gehring, M; Kasraie, S; Kapp, A; Raap, U
Hannover Medical School, Department of Dermatology,
Hannover, Germany
Background: The T-cell cytokine IL-31 has
been described for the induction of severe
Allergy 66, Suppl. 94 (2011): 482642  2011 John Wiley & Sons A/S
Poster Session 52
pruritus and dermatitis in transgenic mice.
Recently, we could show that IL-31 levels
are increased in patients with urticaria and
atopic dermatitis in which eosinophils rep-
resent key target effector cells. However,
the role of IL-31 and its effect in eosino-
phil biology is unknown. Thus, we wanted
to investigate the functional role of IL-31
on eosinophils.
Method: Human peripheral blood eosin-
ophils were isolated by CD16 negative
selection (purity >98%). FACS analysis
was used to detect oncostatin M receptor
(OSMR) and IL-31 receptor A (RA).
Eosinophil responses to IL-31 were moni-
tored by Ca2+ mobilisation (uorometrical
analysis), superoxide anion release (lucige-
nin-dependent chemiluminescence); apopto-
sis (FACS analysis), chemotaxis (modied
Boyden chamber assay) and phospho-spe-
cic Western blotting.
Result: IL-31RA was expressed by human
peripheral blood eosinophils, whereas we
did not nd an expression of OSMR. IL-31
stimulation led to a signicant increase of
intracellular Ca2+ levels and release super-
oxide anions of eosinophils. Further, IL-31
signicantly induced chemotaxis and inhib-
its apoptosis. Finally, IL-31 stimulation led
to an activation of the extracellular signal
regulated kinase (ERK 1/2) and the down-
stream signalling molecule signal transducer
and activator of transcription (STAT)-3.
Conclusion: Together, our data give rst
evidence for a functional role of IL-31 on
human peripheral blood eosinophils, thus
revealing novel implications for IL-31
action.
1301
Involvement of galectin-9 in lung eosino-
philia in patients with eosinophilic pneu-
monia
Katoh, S1; Matsumoto, N2; Shimizu, H1; Obase, Y1;
Fukushima, K3; Hirashima, M4; Oka, M1
1
Kawasaki Medical School, Department of Respiratory
Medicine, Kurashiki, Japan; 2Faculty of Medicine,
University of Miyazaki, Department of Neurology,
Respirology, Endocrinology and Metabolism, Internal
Medicine, Miyazaki, Japan; 3Japan Red Cross Nagasaki
Genbaku Isahaya Hospital, Division of Respiratory
Disease, Nagasaki, Japan; 4Faculty of Medicine,
Kagawa University, Immunology and
Immunopathology, Kagawa, Japan
Background: Although we rst found
galectin-9 (Gal-9) as an eosinophil chemo-
attractant, its role in eosinophilic inam-
mation is still obscure. The purpose of the
present study is to clarify the role of Gal-9
in human eosinophilic pulmonary inam-
mation in comparison with eotaxin
(CCL11).
Method: We measured the levels of Gal-9
and eotaxin in the bronchoalveolar lavage
uid (BALF) of patients with acute and
chronic eosinophilic pneumonia (AEP and
CEP). Furthermore, the biological activi-
ties (chemotaxis and apoptosis) of Gal-9
were compared with those of eotaxin using
interleukin-5-primed or -unprimed eosin-
ophils.
Result: The levels of Gal-9 and eotaxin in
the BALF from patients with AEP and
those with CEP were higher than those
found in the controls. Although there was
little difference in Gal-9 level between
patients with AEP and CEP, the eotaxin
level was signicantly lower in patients
with CEP. In patients with AEP, the eosin-
ophil number correlated well with both the
Gal-9 and eotaxin levels. However, in
patients with CEP, the eosinophil number
only correlated well with the Gal-9 level.
Moreover, the Gal-9 level correlated with
the eotaxin level in patients with AEP, but
there was no signicant correlation
between those levels in patients with CEP.
Anti-Gal-9 antibody treatment strongly
reduces eosinophil chemotactic activity in
the BALF of patients with AEP and in
that of patients with CEP, whereas the
anti-CCR3 (receptor for eotaxin) antibody
strongly reduces this activity in the BALF
of patients with AEP but not in that of
patients with CEP. Furthermore, Gal-9
exhibited both chemotactic and proapop-
totic activities for activated eosinophils,
though eotaxin only exhibited chemotactic
activity.
Conclusion: The present results provide
two possibilities that Gal-9 is involved in
pulmonary eosinophilia in patients with
AEP and CEP, and that Gal-9 exhibits
regulatory functions for activated eosin-
ophils at the site of inammation.
495

1302
Progression to asthma in eosinophilic
bronchitis
Arochena, L1; Fernandez-Nieto, M1; Sastre, B2; del Pozo,
V2; Sastre, J1
1
Fundacion Jimenez Daz, Allergy Department, Madrid,
Spain; 2Fundacion Jimenez Daz, Immunology
Department, Madrid, Spain
Background: Eosinophilic Bronchitis (EB)
is a common cause for chronic cough that,
like asthma, has sputum eosinophilia but,
unlike asthma, it doesnt associate revers-
ible bronchial obstruction or bronchial
hyper-responsiveness (BHR), being BHR a
20% fall in FEV1 after bronchial challenge
with methacholine or histamine.
Method: A total of 17 patients diagnosed
with eosinophilic bronchitis (EB) were
monitored in annual visits during the last
3 years. Inhaled budesonide (400 mg) every
12 h was used as chronic treatment, but
suspended 3 weeks before the visit. A con-
trol of symptoms questionnaire, spiromet-
ric measurements, fraction of exhaled nitric
oxide (FeNO), bronchial challenge with
methacholine and sputum induction exami-
nation by using ow-cytometry were per-
formed every year.
Result: Despite of treatment, 5 patients
(29.41%) still referred chronic cough, while
the other 12 (70.58%) were cough-free.
Spirometric parameters were normal in
100% of patients, and the FeNO mean
was 30.58 20.76 ppb. Sputum eosino-
philia was a constant feature with a mean
of 10.75% 0.11. A total of 16 of 17
patients had no bronchial hyper-respon-
siveness to methacholine (PC20 > 16 mg/
ml) but one had a positive result, with a
PC20 of 5.36 mg/ml.
Conclusion: We present 17 patients diag-
nosed with EB according to Gibson and
Brightling criteria. One of them (5.88%)
had a positive response to bronchial chal-
lenge with methacholine 3 years after the
diagnosis. This suggests the possibility that
sometimes, EB may progress to asthma.
New studies are mandatory to obtain a
more specic knowledge about EB natural
history.
1303
The role of eosinophil in preterm infants
with bronchopulmonary dysplasia
Rha, Y1; Kim, S2; Kim, M3; Choi, S4
1
Kyunghee University Hospital, Department of
Pediatrics, Seoul, Republic of Korea; 2Busan St. Mary
Hospital, Department of Pediatrics, Busan, Republic of
Korea; 3Aigumteo Childrens Hospital, Department of
Pediatrics, Daegu, Republic of Korea; 4Kangdong
Kyunghee University Hospital, Department of
Pediatrics, Seoul, Republic of Korea
Background: Bronchopulmonary dysplasia
(BPD) results in bronchial hyper-respon-
496
siveness and recurrent wheezing. Recent
studies suggested that eosinophil has rele-
vance to innate immune defense and
inammatory process in early life. While
the activation of neutrophil and macro-
phage in pathogenesis of BPD are rather
well studied, very few studies have
addressed the possible participation of
eosinophil. We hypothesized that eosino-
phil may be involved in early phase of
BPD.
Method: Preterm infants were recruited
consecutively and grouped to respiratory
distress syndrome (RDS) or non-RDS.
RDS group was divided to BPD or non-
BPD group then. We investigated blood
eosinophil, serum eosinophil cationic pro-
tein (ECP), eosinophil protein X (EPX),
and urine EPX to determine the relation-
ship between eosinophil activation and
BPD in premature infants prospectively.
The eosinophil count, serum ECP/ EPX
and urine ECP/ EPX were compared.
Spearman rank test was used to analyze
the correlation between serum, tracheal
and urine ECP.
Result: Among total 43 patients recruited,
13 patients were treated with pulmonary
surfactant and mechanical ventilation for
RDS. Six out of 13 patients were diag-
nosed with BPD. There were no difference
of blood eosinophil, serum ECP, EPX, or
urine EPX between RDS and non-RDS
groups. Blood eosinophil, serum ECP,
EPX, and urine EPX were higher in BPD
group than non-BPD group which was
not statistically signicant. Interestingly
serum EPX was signicantly correlated
with pH of arterial blood gas analysis
which was done at discharge. (r = 0.441,
P = 0.012).
Conclusion: Eosinophil activation markers
were higher in BPD group which suggests
the possible role of eosinophil in BPD
although there was no statistical signi-
cance.
1304
Enhanced eosinophilia and autocrine
osteopontin loop in chronic sinusitis with
nasal polyps
Liu, W1; Shi, J2; Li, H2
1
Otorhinolaryngology Hospital, Allergy and Cancer
Center, Guangzhou, China; 2China
Background: Upregulated osteopontin
(OPN) is reported in patients with chronic
sinusitis with nasal polyps (CRSwNP).
However, the regulatory effect of OPN in
the pathogenesis of nasal polyps has not
been understood. This study thus investi-
gated whether OPN production is associ-
ated with enhanced eosinophilia and
clinical severity in patients with CRSwNP.
Allergy 66, Suppl. 94 (2011): 482642  2011 John Wiley & Sons A/S
Poster Session 52 Mast cells and eosinophils
Method: Twenty-ve patients with
CRSwNP and 18 normal controls were
included, and OPN production was deter-
mined by immunohistochemistry (IHC)
and enzyme-linked immuno sorbent assay
(ELISA), respectively. Eosinophil accumu-
lation in nasal polyps and normal controls
were evaluated by immunostaining of
major basic protein (MBP). Co-location of
OPN and MBP in nasal polyps was exam-
ined by double-staining immunouores-
cence. Chemotaxis of eosinophil in nasal
polyps to recombinant OPN was evaluated
through an in-vitro Transwell system.
Results: We observed signicantly
enhanced OPN+ cells and MBP+ cells in
nasal polyps compared with normal con-
trol, which was supported by increased
OPN levels in nasal polyps. OPN+ cells
were found to be positively correlated with
MBP+ cells and clinical severity of nasal
polyps. Furthermore, we provided the
direct evidence that more than 40% OPN
cells were overlapped with MBP+ cells in
nasal polyps by double-staining immuno-
uorescence, and eosinophils in nasal pol-
yps were attracted by recombinant OPN
through an in-vitro Transwell system.
Conclusion: Our results demonstrate that
an enhanced eosinophila and an autocrined
OPN loop in eosinophils in nasal polyps,
and thus provide a possible explanation for
regulatory role of OPN in chemotaxis of
eosinophils and in the pathogenesis of
nasal polyps.
1305
CCR4-acting chemokines contribute to
tissue eosinophilia in Churg-Strauss syn-
drome
Jakiela, B; Sanak, M; Szczeklik, W; Sokolowska, B;
Plutecka, H; Mastalerz, L; Musial, J; Szczeklik, A
Jagiellonian University Medical College, Departament
of Medicine, Krakow, Poland
Background: Churg-Strauss syndrome
(CSS) is a rare systemic vasculitis associ-
ated with asthma and eosinophilic inl-
trates. We studied the contribution of
CCR4-acting chemokines to eosinophilic
inammation in CSS patients, stratied by
the disease activity.
Method: The concentration of IL-5,
CCL17 and CCL22 was measured in sera
and broncho-alveolar lavage uid (BALF)
sampled from 10 patients with active CSS,
9 patients with CSS in remission and 10
healthy subjects (sera only). We also ana-
lyzed mRNA of selected Th2-related genes
by quantitative PCR in BALF cells.
Result: The concentration of serum IL-5
and CCR4-acting chemokines were signi-
cantly increased in active CSS as compared
to patients in disease remission (CCL17:
699 [3511502] versus 257 [171414] pg/ml;
Poster Session 52 Mast cells and eosinophils
CCL22: 1095 [6092855] versus 420 [192
520] pg/ml; P < 0.05), and healthy con-
trols (CCL17: 258 [193350] pg/ml,
P < 0.05; CCL22: 654 [542780] pg/ml,
P = 0.06). Serum levels of these chemo-
kines correlated with clinical symptom
scores (BVAS) and blood eosinophilia. In
BALF from active-CSS, both eosinophil
counts and IL-5 concentration were signi-
cantly increased (IL-5: 1.26 [0.65.4] versus
0.54 [< 0.30.62] pg/ml in remission,
P = 0.01). CCR4-acting chemokines were
elevated in BALF from 4 patients with
exacerbation and 1 in remission. We also
observed an increase in cellular abundance
of Th2-type transcripts (e.g. IL4, IL5,
CCR4) in BALF from patients with active
disease.
Conclusion: Overexpression of CCR4-act-
ing chemokines contributes to the lung
eosinophilia and severity of CSS symptoms
by recruitment of Th2 lymphocytes and eo-
sinophils to the site of CSS-related inam-
mation.
1306
Asymptomatic toxocariasis as a cause of
eosinophilia: a report of two cases
Vassilopoulou, I1; Vassalou, E2; Panos, G3; Trigka, M1
1
Patras University General Hospital, Paed Allergy Unit,
Department of Paediatrics, Patras, Greece; 2School of
Public Health, Department of Parasitology, Entomology
and Tropical Diseases, Athens, Greece; 3Patras
University General Hospital, Infectious Diseases Unit,
Dept of Internal Medicine, Patras, Greece
Background: Eosinophilia (absolute eosino-
phil count >500/ll) is not an uncommon
nding in clinical practice and can occur in
a wide variety of conditions. We report
two cases admitted to the paediatric allergy
unit for eosinophilia investigation.
Case 1: A 9-year-old girl was admitted
with 25% eosinophils of total WBC (abso-
lute eosinophil number 2000 cells/ll) found
at routine testing. Total serum IgE was
>1000 IU/ml.
Case 2: An 11-year-old boy was admitted
with 20% eosinophils of total WBC (abso-
lute eosinophil number 830 cells/ll) also
found at routine testing. Total serum IgE
was 2780 IU/ml.
Both children had no history of any ato-
pic, immunologic, gastrointestinal or auto-
immune disease, neither travel history or
medication use. They had no any symptoms
or complaints and their general condition
and physical examination were normal. The
laboratory investigation (Hct, Hb, CBC
smear, ESR, CRP, routine liver and renal
function tests, IgG, IgA, IgM, C3, C4,
ANA, antiDNA, stool microscopic and cul-
ture examination), the imaging examination
(chest-X ray, abdominal ultrasound), and
the skin-prick tests to common allergens
Allergy 66, Suppl. 94 (2011): 482642  2011 John Wiley & Sons A/S
were all within normal. Antibodies against
toxocara were detected in the serum of both
children by enzyme-linked immunosorbent
assay (ELISA) employing extracts of larval
antigens of toxocara canis. The patients
were treated with a 10-day course of alben-
dazole and the eosinophil count was found
normal at the end of the treatment and at
the 6-month and 1-year follow-ups.
Conclusion: Toxocariasis should be strongly
considered in asymptomatic patients with
eosinophilia even in the developed world,
because many cases might be misdiagnosed
as hypereosinophilic syndrome leading to
inappropriate treatment. Treating toxocari-
asis in asymptomatic children with eosino-
philia seems prudent since the role of
eosinophil as protective or inammatory
agent in parasitic infections is poorly under-
stood and remains debatable.
1307
Eosinophilic dermatosis in B-cell chronic
lymphocytic leukaemia: production of
interleukin-3 and 5 by reactive T lympho-
cytes
Posa, M1; Matthes, T2; McKee, T3; Masouye, I4; Laffitte,
E4; Simon, H5; Seebach, J1
1
Geneva University Hospital, Division of Clinical
Immunology and Allergy, Geneva, Switzerland;
2
Geneva University Hospital, Division of Haematology,
Geneva, Switzerland; 3Geneva University Hospital,
Department of Pathology, Geneva, Switzerland;
4
Geneva University Hospital, Division of Dermatology,
Geneva, Switzerland; 5Bern University Hospital,
Institute of Pharmacology, Bern, Switzerland
Background: Eosinophilic dermatosis with
insect bite-like or ekzematiform prurigin-
ous skin lesions has been rarely described
in association with hematologic diseases
such as chronic lymphocytic leukemia.
Blood eosinophilia can be frequently found
in patients with lymphoma, in the presence
of a clonal T lymphoid population, in con-
trast malignant haematopathies of the
B-cell lineage are only exceptionally associ-
ated with peripheral blood eosinophilia.
Method: Here, we describe the case of a
71-year-old man who reported itching,
eczema on his scalp, upper arms and back,
as well as fatigue and dyspnoea. Peripheral
blood eosinophils were elevated to 3000/
mm3 (32%), whereas haemoglobin, lym-
phocyte counts, and IgE levels were nor-
mal. No allergies or parasitic infections
were found in the workup for eosinophilia,
his treatment for hypertension was chan-
ged from an angiotensin-converting
enzyme inhibitor with a beta-blocker with-
out amelioration of the skin lesions and
eosinophilia. A CT scan revealed lymph-
adenopathy and splenomegaly, as well as
multiple pulmonary nodules.
Result: Histologically, a skin biopsy showed
a perivascular and interstitial eosinophilic
inltrate, as well as a population of T lym-
phocytes. A diagnosis of CD19+, CD5+,
CD23+, CD38+, CD200+, CD20dim,
CD10-, FMC7-, kdim B-cell chronic lym-
phocytic leukaemia (B-CLL) was made by
phenotypic analysis of lymphocytes in the
peripheral blood, bone marrow biopsy and
lymph node biopsy. T and B cell receptor
analysis of peripheral blood, bone marrow,
skin and lymph node samples by molecular
biology revealed a monoclonal B lymphoid
population without evidence of T cell clo-
nality. In vitro stimulation assays demon-
strated the production of IL-3 and IL-5 by
reactive T lymphocytes but not by the clonal
B lymphocytes. The patients symptoms as
well as blood eosinophilia responded well to
moderate doses of corticosteroids (25 mg of
prednisone/day).
Conclusion: This is the rst case showing a
link between a reactive T lymphoid popula-
tion and blood eosinophilia in a patient suf-
fering from subclinical B-CLL. A clinical
presentation of ekzematiform pruriginous
skin lesions associated with blood eosino-
philia without apparent cause demands an
active search for hematologic disorders
which are the third cause of hypereosino-
philia, after allergic and parasitic diseases.
1308
AntiCCR3 antibody inhibits oral food
allergen induced intestinal eosinophilic
inflammation in a murine model
Song, D; Yoo, Y; Shim, M; Choung, J
College of Medicine, Korea University, Department of
Pediatrics, Seoul, Republic of Korea
Background: Although the role of eosin-
ophils in eosinophilic gastrointestinal dis-
eases (EGIDs) is not fully understood, they
are believed to be one of the principal
effector cells inducing gastrointestinal tis-
sue injury and disease pathogenesis. CCR3
is a chemokine receptor predominantly
expressed on eosinophils and involved in
the recruitment of eosinophils into inam-
matory site. This study was aimed to inves-
tigate the effect of targeting CCR3 on
intestinal eosinophilic inammation and
tissue injury.
Method: Using a murine model, we ana-
lyzed the effects of anti-CCR3 antibody on
oral food allergen induced eosinophilic
inammation in the intestinal mucosa and
intestinal tissue injury compared with that
of control antibody.
Result: Administration of antiCCR3 anti-
body resulted in the abolition of eosinophils
from intestinal mucosa, which was associ-
ated with signicantly reduced numbers of
peripheral blood eosinophils, but was not
associated with signicant changes in the
numbers of bone marrow eosinophils. Mice
497

administered antiCCR3 antibody had lower
level of intestinal epithelial proliferation,
intestinal villous crypt height change, and
weight loss.
Conclusion: AntiCCR3 antibody inhibited
oral food allergen induced eosinophilic
inammation in the intestinal mucosa,
which was associated with reduced intesti-
nal tissue injury and weight loss. Targeting
of CCR3 may be a potential therapeutic
intervention for EGIDs and other eosino-
phil-mediated diseases.
1309
Bone marrow derived mast cells from
microRNA-155 knockout mice show
impaired stem cell factor-mediated
migration
Radinger, M; Malmhall, C; Sjostrand, M; Lotvall, J
University of Gothenburg, Krefting Research Centre/
Sahlgrenska Academy, Gothenburg, Sweden
Background: MicroRNAs (miR) are a
growing class of small noncoding RNAs
that regulate gene expression by targeting
mRNAs. Thus, these molecules are emerg-
ing as important modulators in cellular
immune responses. Chronic allergic inam-
mation is characterized by increased mast
cell inltration in the affected tissues. Mast
cell burden in these tissues is dependent on
migration of mast cells/mast cell precursors
into target tissue. Such processes are in
part regulated by stem cell factor (SCF),
the ligand for the growth-factor receptor
KIT. In this study we investigated the role
of microRNA-155 (miR-155) in stem cell
factor (SCF) -mediated mast cell chemotac-
tic responses.
Method: Mouse bone marrow derived mast
cells (BMMCs) were developed from wild
type mice (WT) and mice decient in miR-
155 (miR-155 KO mice). Mature BMMCs
(46 weeks of growth) were used for exper-
iments. In vitro chemotaxis assays were
performed using a Transwell system and
migrated cells were collected and counted
by microscopy.
Result: No difference was shown in mast
cell development or maturation of BMMCs
from miR-155 KO mice compared to WT
derived BMMCs. In contrast, BMMCs
derived from miR-155 KO mice showed a
signicantly decreased SCF-mediated che-
motaxis compared to WT derived BMMCs
(2.3 0.6 x 103 cells versus 8.9 1.6
103 cells P < 0.01).
Conclusion: These data provide evidence
that miR-155 is involved in the regulation
of SCF-mediated mast cell migration.
Thus, targeting miR-155 may provide a
mechanism for modulating mast cells
migrational pathways in the allergic inam-
matory response.
498
1310
IL-33 induced mast cells activation lead-
ing to Th17 differentiation in ovalbumin-
challenged mice
Cho, K1; Suh, J1; Woo, S1; Cho, Y2
1
School of Medicine, Ewha Womans University,
Department of Microbiology, Seoul, Republic of Korea;
2
Ewha Womans University Hospital, Allergy and
Clinical Immunology, Departmeny of Internal medicine
Department, Seoul, Republic of Korea
Background: Allergic asthma is character-
ized by inltration of eosinophils, elevated
Th2 cytokine levels, airway hyper-respon-
siveness (AHR), and IgE. Aside from eo-
sinophils, mast cells, basophils, and a
variety of cytokines are also involved in
the development of allergic asthma. IL-33
can promote the pathogenesis of asthma
by expanding Th2 cells and blocking IL-33
signaling, thereby suppressing the asth-
matic response in eosinophils, basophils
and mast cells.
Methods: In order to determine the out-
come of inammation involving IL- 33 in
mouse allergic asthma model, we induced
airway inammation via ovalbumin
(OVA)-challenge in Balb/c mice and eosin-
ophil-decient mice and analyzed the cells
recruited to the lungs and cytokine pro-
les.
Results: We demonstrated that IL-33 acts
directly on mast cells to generate IL-1b
and IL-6, and that such interactions
between IL-33 and mast cells enhanced
Th17 differentiation, which may induce
neutrophil-dominant airway inammation
under OVA-challenge conditions. IL-33
was upregulated in either WT mice or
eosinophil-decient mice under OVA-
induced airway inammation, and these
effects were accompanied by an increase in
ST2 in the lung tissue. In particular,
DdblGATA mice evidenced neutrophilic
airway inammation and increased per-
centages of mast cells relative to the OVA-
challenged WT mice.
Conclusions: These results indicate that
without the inuence of eosinophils, mast
cells predominantly affect immune reac-
tions toward Th17 responses via IL-33
stimulation.
1311
Immunoglobulin free light chains syner-
gistically enhance IgE-mediated mast cell
activation
Blokhuis, B; Thio, M; Redegeld, F
Utrecht University, Pharmacology, UIPS, Fac. Science,
Utrecht, The Netherlands
Background: Mast cells are key effector
cells in allergic diseases. The most promi-
nent pathway to activate mast cells is via
crosslinking of IgE bound to the FceRI
Allergy 66, Suppl. 94 (2011): 482642  2011 John Wiley & Sons A/S
Poster Session 52 Mast cells and eosinophils
resulting in degranulation and mediator
release. In previous work, we have shown
that activation via immunoglobulin free
light chains (IgLC) may be an alternative
pathway of antigen-specic mast cell acti-
vation. In recent studies we found that
local or circulating IgLCs are increased
and may be of importance in triggering
mast cell activation in (non-IgE mediated)
inammatory diseases such as non-atopic
asthma, inammatory bowel disease, food
allergy, rhinitis and rheumatoid arthritis.
Since we found that IgLC are also often
increased in allergic (IgE-driven) diseases,
we investigated if IgLC could also inu-
ence IgE-mediated mast cell activation.
Method: Primary cultured bone marrow-
derived mast cells and the mast cell lines
MC/9, CFTL15 and RBL-2H3 were stimu-
lated by IgE in presence of various concen-
trations of IgLCs. Mast cell activation was
monitored by the release of beta-hexosa-
minidase and IL-6 production.
Result: Mast cell activation by IgE/FceRI
was greatly enhanced when simultaneously
IgLC were crosslinked on the mast cell sur-
face. When IgE and IgLC was titrated to
concentrations which induced only limited
to no mast cell activation (<5% beta-hex-
osaminidase release) combined crosslinking
of IgE plus IgLC resulted in a synergistic
activation of all tested mast cells. Both
degranulation and IL-6 production were
greatly increased in such combinations of
IgE and IgLC. Mast cell activation was
completely inhibited by syk kinase inhibi-
tors.
Conclusion: This study indicates that com-
bined crosslinking of IgE and IgLC results
in synergistic activation of mast cells. Our
results suggest that production of IgLC in
IgE-mediated diseases may be of clinical
relevance for the induction of allergic
symptoms.
1312
Functional expression of TRPV4 cation
channels in human mast cell line
Kim, W1; Nam, J2
1
Dongguk University Ilsan Hospital, Asthma & Allergy,
Kyunggi, Republic of Korea; 2Dongguk University
College of Medicine, Physiology, Kyungju, Republic of
Korea
Background: Mast cells are activated not
only by the specic allergens but also by
various non-specic stimuli, which might
induce physical urticaria.
Method: In this study we investigate the
functional expression of temperature sensi-
tive TRPV subfamily in human mast cell
line (HMC-1) using whole-cell patch clamp
techniques. The temperature of perfusate
was raised from the room temperature
(RT, 2325J) to moderately high (MHT,
Poster Session 52 Mast cells and eosinophils
3739J) for activating TRPV3/4, high
(HT, 4446J) for TRPV1, or very high
temperature (VHT, 5355J) for TRPV2.
Results: The membrane conductance of
HMC-1 was increased by MHT and HT in
about 50% (21 out of 40) of the tested
cells, and the I/V curves showed weak out-
ward rectication. VHT-induced current
was 10-fold larger than those induced by
MHT and HT. The application 4a-phorbol
12,13-didecanoate (4aPDD, 1 lM), an acti-
vator of TRPV4, induced weakly outward
rectifying currents similar to those induced
by MHT. However, camphor (TRPV3 ago-
nist) or capsaicin (TRPV1 agonist) had no
effect. RT-PCR analysis of HMC-1 dem-
onstrates the expression of TRPV4 as well
as the strong expression of TRPV2. The
[Ca2+]c of HMC-1 cells was also increased
by MHT or by 4aPDD.
Summary: Our present study indicates that
HMC-1 cells express Ca2+-permeable
TRPV4 channels in addition to the previ-
ously reported expression of TRPV2 with
higher threshold of activating temperature.
1313
Neurotrophins in mast cells are equally
involved in allergic and non-allergic rhini-
tis
Gelincik, A1; Aydin, F2; Ozarman, B3; Ozcan, B2;
Demirturk, M1; Colakoglu, B1; Erden, S4; Dal, M1;
Buyukozturk, S1
1
Istanbul University, Istanbul Faculty of Medicine,
Department of Internal Medicine, Division of Allergy,
Istanbul, Turkey; 2Istanbul University, Istanbul Faculty
of Medicine, Department of Medical Biology, Istanbul,
Turkey; 3Istanbul University, Istanbul Faculty of
Medicine, Department of Biophysics, Istanbul, Turkey;
4
Istanbul University, Istanbul Faculty of Medicine,
Department of Internal Medicine, Istanbul, Turkey
Background: The role of neurotrophins in
the upper-airway pathophysiology of aller-
gic rhinitis (AR) has been well studied, but
it has not been evaluated in non-allergic
rhinitis (NAR). According to the prelimin-
ary data of our study, the neurotrophins
and their receptors were found to be
equally important in NAR and AR and
interestingly mast cells were increased in
number in NAR. In our present study we
aim to evaluate whether b-NGF is
expressed in mast cells in both AR and
NAR patients.
Method: Seventeen persistent moderate-
severe AR patients (mean age: 29.7
11.96) diagnosed according to ARIA
guidelines with monoallergy to house-dust
mite were included in the study. Fourteen
NAR patients (mean age 29.3 10.62)
and sixteen healthy controls (29.9 11.57)
were also studied. Patients with infectious,
atrophic, NARES (non-allergic rhinitis
with eosinophilic syndrome), iatrogenic, or
hormonal rhinitis and the ones symptom-
Allergy 66, Suppl. 94 (2011): 482642  2011 John Wiley & Sons A/S
atic due to the anatomic defects were not
included in the study. Nasal biopsies were
taken from the posterior part of the infe-
rior turbinate from all the study subjects.
Pan-neurotrophin receptor p75, tyrosine
kinase A (trkA) and b-nerve growth factor
(b-NGF) were assessed with immunouo-
rescence assay in nasal biopsy specimens.
Mast cell tryptase activity was detected im-
munohistochemically in corresponding for-
malin-xed, parafn-embedded nasal
tissues. Mast cell tryptase positive cells had
Brown stained cytoplasm. The b-NGF in
mast cells was detected by two different
uorescence-labelled antibodies. b-NGF
was labelled by anti-goat IgG-FITC while
mast cell tryptase activity was detected by
labelling with donkey anti-mouse IgG rho-
damine.
Result: When the patients with AR were
compared to the controls, b-NGF positive
mast cells were found to localize abun-
dantly in the basal layer of the epithelium,
in the connective tissue beneath the epithe-
lium and in the perivascular area. Whereas
in patients with NAR, b-NGF positive
mast cells were localized commonly in the
same areas like AR in addition in the basal
of the mucosal glands.
Conclusion: Mast cells were found to
express b-NGF equally in AR and NAR.
Therefore we propose that mast cells do
play a role in the pathogenesis of NAR as
importantly as AR.
1314
Antiapoptotic Bfl-1 is the major effector
in activation-induced human mast cell
survival
Lyberg, K1; Ekoff, M1; Harvima, I2; Nilsson, G1
1
Karolinska Institutet, Department of Medicin, Centre
for Allergy Research, Stockholm, Sweden; 2Kuopio
University Hospital, Department of Dermatology,
Kuopio, Finland
Background: Allergy and asthma are typi-
cal examples of diseases where the number
of mast cells in the affected tissue increases
and a correlation between the number of
mast cells and the severity of the symptoms
has been reported. In allergy, mast cells
are known for their ability to mediate IgE-
dependent responses, where aggregation of
FceRI leads to the release of mast cell
mediators causing allergic symptoms.
Following activation mast cells have the
ability to survive, regranulate and be
re-stimulated and thereby continue to pro-
mote the allergic symptoms. A fundamen-
tal question in mast cell biology is how
this survival is mediated. We have previ-
ously shown that mouse mast cells decient
in the anti-apoptotic bcl-2 family member
A1 do not exhibit activation-induced sur-
vival upon FceRI crosslinking. In this
study we have investigated the human
homologue B-1 and its importance for
activation-induced mast cell survival.
Method: Immunohistochemestry, QPCR
and owcytometry
Result: By immunohistochemicalstaining
for B-1 in skin biopsies from allergen
challenged skin we observed a clear upreg-
ulation of B-1 in skin residing mast cells
in comparison with control biopsies. Major
inhibition of activation-induced mast cell
survival was obtained using b-1 siRNA
compared to a non-targeting siRNA pool
transfected into cord-blood derived mast
cells which were analyzed by QPCR. Acti-
vation-induced survival was measured
using PI/AnnexinV staining and analyzed
by FACS. It proved to be conserved when
the anti-apoptotic proteins Bcl-XL, Bcl-2,
Bcl-w and Mcl-1 were inhibited by the
inhibitors ABT-737 and roscovitine, indi-
cating a minor role for these anti-apoptotic
Bcl-2 family members in activation-induced
mast cell survival.
Conclusion: Taken together, our results
highlight B-1 as a major effector in acti-
vation-induced human mast cell survival.
We therefore suggest that it would be of
great interest to identify potential binding-
partners of B-1 in mast cells and to evalu-
ate B-1 as a target for treatment of aller-
gic diseases.
1315
The effect of N-acetylcysteine in a rat
model of allergic rhinitis
Guibas, G1; Spandou, E1; Meditskou, S2; Vyzantiadis,
T3; Anogianakis, G1
1
Aristotle University of Thessaloniki, Laboratory of
Experimental Physiology, Thessaloniki, Greece;
2
Aristotle University of Thessaloniki, Laboratory of
Histology-Embryology, Thessaloniki, Greece; 3Aristotle
University of Thessaloniki, Laboratory of Microbiology,
Thessaloniki, Greece
Background: The pathophysiologic mecha-
nism of allergic rhinitis is initiated and
kept in motion through a process that is
depended upon a variety of redox-sensitive
proinammatory mediators. However, even
though redox disturbances are believed to
be a hallmark of inammation, little is
known of the effect of oxidant-antioxidant
imbalance to the pathophysiology of aller-
gic rhinitis.The aim of this study was to
investigate the relation of oxidative stress
and allergic rhinitis, through the utilization
of the potent antioxidant N-acetylcysteine
(NAC) in a rat model of allergic rhinitis,
and the evaluation of its action on specic
markers of inammation.
Method: NAC was administered to ovalbu-
min (OVA)-sensitized rats prior to intrana-
sal challenge with OVA. The degree of
nasal mucosal inammatory inltration,
the state of epithelial mucus cells (goblet
499

cells), the expression of mucosal redox-sen-
sitive enzymes (inducible Nitric Oxide Syn-
thase/iNOS and Cyclooxygenase 2/COX-2)
and the levels of a key proinammatory
mediator (Tumor Necrosis Factor-a/TNF-
a) in the blood serum were evaluated.
Result: Intranasal challenge of the sensi-
tized rats lead to inammatory inltration
of the nasal mucosa, hyperplasia of goblet
cells, induction of iNOS and COX-2 muco-
sal expression and elevation of TNF-a lev-
500
els in the blood serum. NAC exerted dose-
depended action that was more potent in
the lower dose, regarding the suppression
of mucosal eosinophilia and goblet cells
hyperplasia as well as the inhibition of
iNOS expression and TNF-a serum levels.
The role of COX-2, in the inammatory
process appeared to be divergent and its
expression was not distinctly modulated by
NAC.
Allergy 66, Suppl. 94 (2011): 482642  2011 John Wiley & Sons A/S
Poster Session 52 Mast cells and eosinophils
Conclusion: Redox balance is involved in
the pathophysiology of allergic rhinitis in
rats and NAC can potentially suppress the
allergen-induced nasal inammatory cas-
cade. The investigation of the role of oxi-
dative stress in the atopic condition can
help in the evaluation of the therapeutic
potential of antioxidant substances in the
treatment of allergic diseases.

Novel clinical and basic aspects of allergen immunotherapy
1316
Malignancy and specific allergen
immunotherapy the results of a case
series
Kinaciyan, T1; Jalili, A1; Stingl, G1; Woehrl, S1;
Moritz, K2
1
Austria; 2Dep. Dermatology, Div. Immunology, Allergy
and Infectious Diseases (DIAID), Vienna, Austria
Background: Specic immunotherapy with
allergen is the only causative treatment for
IgE-mediated allergies such as stinging
insect allergy or hay fever and works by
the induction of blocking antibodies and
regulatory T lymphocytes.
Objective: Does a hypothetical obstruction
of tumor surveillance presupposing the
induction of regulatory T-cells by immuno-
therapy really justify detaining immuno-
therapy to oncologic patients as suggested
by recent guidelines?
Methods: We report 6 patients (4 female, 2
male) suffering or having suffered from
stage 1 cancer (4 melanomas, 1 lung
cancer, 1 breast cancer) and concomitant
IgE-mediated allergy. Four of them had a
history of severe anaphylactic reactions to
the insect yellow-jacket, the 5th suffered
from allergic rhinoconjunctivitis to dust
mites, the 6th to grass/rye pollen.
Results: Between 2004 and 2010, subcuta-
neous immunotherapy was safely per-
formed in 5 patients without signs of
tumor reactivation. The cancer in 2 of
them was diagnosed immediately after SIT
had been initiated and in another 2 the
active cancer phase had already passed by
years before, the 5th suffered from a
relapse around the time of the initiation of
immunotherapy. At the time of the writing
of this manuscript, 4 of them had already
concluded 3 years of treatment, in another
one more than 1 year. The melanoma in
the 6th patient was diagnosed 5 months
after reaching the maintenance dose. The
immunotherapy with grass/rye pollen was
aborted in this patient based on current
guidelines.
Conclusion: SIT was safely administered in
patients concomitantly suffering from IgE-
mediated allergy and lower stage cancer.
Allergy 66, Suppl. 94 (2011): 482642  2011 John Wiley & Sons A/S
Poster Session 53
1317
Development of a multi-dose subcutane-
ous product for flexible patient treatment
Oluoma, L; Batten, T; Williams, T; Skinner, M
Allergy Therapeutics, Development, Worthing, United
Kingdom
Background: Multi-dose formulations for
subcutaneous route of administration are
advantageous particularly in providing the
clinician with exibility in dosing. Multi-
dose formulations must contain antimicro-
bial agents to protect them from potential
microbial contamination during multiple
withdrawals from vials. A comprehensive
package of data which supports multiple
product intervention is presented including,
but not limited to, in-use, photo-stability
and preservative efcacy studies.
Method: Over the past decade, there has
been an increase in the number of commer-
cial protein products and a small amount
has been marketed in a multi-dose congu-
ration. The paucity of multi-dose protein
products may be related in part to the dif-
culty of selecting appropriate preserva-
tives. In-Use and preservative efcacy
study database can support optimal preser-
vatives for a multi-dose formulation, which
demonstrates that products are free from
microbial contaminants after every inter-
vention tested and that protein stability is
correlated with preservative concentration.
Result: Selection of the optimal preserva-
tive(s) is dependent on a number of fac-
tors. Preservatives need to be compatible
with the route of administration and be
effective against various strains of fungi
and bacteria including, but not limited to,
Candida albicans, Escherichia coli, Gram-
negative Bacteria and Salmonella spp. The
effect of the preservative on protein stabil-
ity is a major concern. The preservative
used in a number of sterile products has
been investigated and found to be compati-
ble for use in subcutaneous route of
administration. Our extensive In-Use and
photo-stability studies conrm products
are insensitive to environmental challenges
such as light reactivity.
Conclusion: A comprehensive series of sta-
bility studies and preservative efcacy anal-
yses provides a rationale for exibility in
dosing of multi-dose formulations for sub-
cutaneous use.
1318
Peculiarities of specific immunotherapy
for patients with pollinosis and second-
ary immunodeficiency syndrome
Sizyakina, L; Yagubyan, R
Rostov State Medical University, Department of Clinical
Immunology and Allergology, Rostov-on-Don, Russian
Federation
A specic immunotherapy (SIT) is usually
used to treat pollinosis, however the SIT
efciency is not enough in majority of
cases, especially among patients with the
secondary immunodeciency syndrome.
The use of immunotropic medication to
correct the secondary immunodeciency
syndrome (SIS) in such cases is of special
interest, however it is worth mentioning
that the use of the specied medication
while SIT is disputable.
Method: Used 48 patients, divided into
two subgroups. One of them got SIT with
preventive introduction of isoprinosine
(500 mg three times a day) within 10 days
and a refresher course of treatment in two
weeks. The patients of the second sub-
group got only standard SIT. All patients
often had ARVI before the treatment
(9.35 1.2 per year lasting about
13.7 0.12 days), which signicantly lim-
ited the SIT capabilities. The group receiv-
ing isoprinosine had 6.8 0.7 ARVI
episodes per year; average duration was
8.4 1.3 days after the rst course.
Results: While analyzing the efciency of
the carried out therapy on Dushein score
we have found a decrease in duration of
allergic manifestations in the group of
patients receiving isoprinosine (3.1 1.1
before treatment and 3.7 0.9 after the
course), although in the control group it
was 3.2 1.3 and 3.4 0.7 accordingly.
The immune status was characterized by a
more apparent positive dynamics in rela-
tion with CD3 lymphocytes (69.6 3.5%
and 54.9 2.5% accordingly, if compared
to the control group), CD4 cells
(41.5 2.3 and 34.2 3.1 accordingly),
normalization of the helper-suppressor cell
ratio, higher content of CD16, providing
cytotoxic effect (14.8 2.3 and
10.5 3.1 accordingly). It is worth paying
attention to the more expressed dynamics
of the correlation between Th1 and Th2
with the Th1 dominance.
501

Conclusion: Summing up the obtained
data, it is worth mentioning that prelimin-
ary isoprinosine introduction contributes
to elimination of the SID syndrome, typi-
cal of patients with intermitting allergic
rhinitis against often ARVI, and contrib-
utes to a better efciency of the carried out
specic immunotherapy.
1319
Effects of immunotherapy with birch pol-
len on food-allergen specific antibodies
Subbarayal, B1; Mobs, C2; NW, d3; Ebner, C4; Reider,
N5; Gerth van Wijk, R3; Pfutzner, W2; Bohle, B1
1
Medical University of Vienna, Department of
Pathophysiology and Allergy Reserach, Vienna, Austria;
2 by SIT. However, such data have been
Philipps University, Department of Dermatology and
Allergology, Marburg, Germany; 3Erasmus MC-
University Medical Center, Department of Allergology,
Department of Internal Medicine, Rotterdam,
Netherlands; 4Allergy Clinic Reumannplatz, Vienna,
Austria; 5Department of Dermatology, University Clinic
Innsbruck, Austria
Background: Cross-reactivity between the
major birch pollen allergen, Bet v 1, its
homologue in apple (Mal d 1) and in
hazelnut (Cor a 1) causes food allergy in
more than 70% of birch pollen-allergic
patients. Specic immunotherapy (SIT)
with birch pollen (BP-SIT) is not effective
to cure birch pollen-related food allergy in
all patients. BP-SIT induces allergen-spe-
cic IgG4 blocking antibodies (Ab), tistically signicant difference in frequency
which compete with IgE Ab for allergen
binding.
Aim: We sought to analyze the levels and
blocking capacity of IgG Ab specic for
Bet v 1 and its related food allergens.
Methods: Sera from 48 BP-allergic patients
were collected before and after BP-SIT.
IgE and IgG4 specic for Bet v 1, Mal d 1
and Cor a 1 were quantied by Immuno-
CAP. All BP-SIT-treated patients devel-
oped higher levels of IgG4 Abs specic for
Bet v 1 than to its homologous food aller-
gens. Around 30% of the patients failed to
develop food-specic IgG4 Abs. The block-
ing capacity of sera obtained before and
after BP-SIT was measured by IgE-facili-
tated allergen binding (FAB) assay, serum-
facilitated allergen presentation (S-FAP)
assay and basophil activation test (BAT)
assay.
Results: BP-SIT-induced allergen-specic
IgG4 Abs effectively reduced (i) IgE-bind-
ing to Bet v 1, (ii) basophil activation by
Bet v 1, and (iii) Bet v 1-induced T cell
proliferation. However, BP-SIT-induced
IgG4 Abs were less effective to block food
homologues.
Conclusion: In summary, BP-SIT does not
induce blocking food-specic IgG4 Abs in
all patients which may explain its limited
clinical efcacy on birch pollen related
food allergy.
502
Poster Session 53 Novel clinical and basic aspects of allergen immunotherapy
1320
Frequency and intensity of bronchial
hyperreactivity in time of colds in
patients with allergic asthma on specific
immunotherapy
Neziri-Ahmetaj, L
UCC, Physiology and Immunology, Prishtina, Albania
Viral infections augment immediate and
late allergic responses in the lungs of
patients with allergic asthma. Certain
viruses that typically exacerbate asthma
have been noted to induce release of the
cytokine interleukin-11(IL-11); which is
associated with AHR.
Immunotherapy has been used exten-
sively for 80 years as a specic treatment
of allergy.
Moreover, allergen vaccination in mono-
sensitized young children was found to
reduce the onset of new sensitizations.
More studies are however needed to deter-
mine how allergen vaccination may modify
the allergic disease or impair progression
to asthma.
Hypothesis: The decrease of serum level of
IL3 and IL11 at the patients with allergic
asthma, who are on immunotherapy, result
with less frequency and intensity of BHR
in presence of viral infections, than at the
patients who are not on immunotherapy.
Goals: To determine if there exists an sta-
number of producing cells, and increased
of cold conditions, the serum level of IL3
and IL4, the frequency and intensity of
bronchial hyper-reactivity, number of eo-
sinophils and neutrophils in DBP, and the
level of IgE in the serum of patients with
allergic asthma that are on immunotherapy
compared to patients with allergic asthma
that are not on immunotherapy for the
period of one year of observation.
Examines and Methods: This is a prospec-
tive, comparative clinical research that will
be done in the University Clinical Hospital
in Pristine and in cooperation with Special-
ized Allergologic Center Yili in Pristine
(Kosovo). 60 patients of both sexes that
have allergic asthma will be included in
this study and they will be divided into
two groups. In the examined group there
will be 30 examinees that will receive
immunotherapy, while in the control group
there will be 30 examinees that will be trea-
ted with other medications but not with
immunotherapy (GINA proposal). Exami-
nees of the examined group will be treated
with subcutaneous specic immunotherapy
(SCIT) each. The level of IL3 and IL11
will be determined from the peripheral
blood of both groups of examinees
through ELISA test. Determination of IL3
and IL11 will be done every 3, 6, 9 and
12 months, and the frequency of cold con-
ditions in these patients will be measured.
Allergy 66, Suppl. 94 (2011): 482642  2011 John Wiley & Sons A/S
1321
Early immunoregulatory changes induced
by rush-specific immunotherapy
Glowania, A1; Pfaar, O1; Hormann, K2; Klimek, L3
1
University Hospital Mannheim, Center for Rhinology &
Allergology / ENT-Department, Wiesbaden, Germany;
2
University Hospital Mannheim, ENT-Department,
Mannheim, Germany; 3Center for Rhinology &
Allergology, Wiesbaden, Germany
Background: Specic immunotherapy (SIT)
is the only pathophysiologically-based
option to treat airway diseases. Usually,
SIT includes an initiation (build-up) phase
and a continuation phase of up to 3 years.
So far, little is known about early induc-
tion of immunoregulatory changes induced
published for venom immunotherapy.
Method: PBMC and monocytes were iso-
lated, and serum samples were taken
before and at the end of a ve day build-
up phase from 12 patients allergic to pollen
and house dust mites. Expression level of
tolerogenic markers were analyzed by ow
cytometry and cell proliferation after stim-
ulation with the respective allergen. Protein
levels of INF-gamma, IL-4 and IL-10 were
measured by ELISA.
Result: We observed signicantly elevated
IL-10 production of CD3(+)-T-cells and
of CD4(+)-T-cells and of IFN-gamma
concerning the amount secreted and the
IL-10 serum levels and IL-10 secretion of
monocytes. Signicant decreases were
observed for proliferative response of
PBMC to the respective allergen.
Conclusion: These data indicate that in
patients allergic to aeroallergens similar to
earlier studies with insect venom allergic
subjects, SIT induces a rapid induction of
protective mechanisms against allergic reac-
tions even in a very short build-up phase
of only 5 days. The described changes in
cytokine expression may be responsible for
subsequent increases in allergen-specic
IgG and decreases in IgE production.
1322
A phase I clinical trial with SCIT vaccines
from timothy grass pollen extracts
according to the new EMA guidelines
Sola, J1; Sanchez, V1; Madariaga, B2; Landeta, A2;
Asturias, J2; Martinez, A2; Alvarez-Cuesta, E1
1
Ramon y Cajal University Hospital, Allergy Service,
Madrid, Spain; 2Bial-Aristegui, Research and
Development Department, Bilbao, Spain
Background: There are only a few pub-
lished studies covering phase I clinical tri-
als of allergen immunotherapy and most of
them belong to sublingual preparations. In
June 2009 a new EMA Guideline concern-
ing clinical development of products for
specic immunotherapy for the treatment
Poster Session 53 Novel clinical and basic aspects of allergen immunotherapy
of allergic diseases came into force
(CPMP/EWP/18504/2006) recommending
that early studies should be done under a
phase I format to test different doses to
provide preliminary data on safety and tol-
erability with regard to the maximum tol-
erated dose and a suitable dose escalation
scheme.
Method: A single centre phase I rando-
mised double blind placebo controlled
study of subcutaneous immunotherapy in
adult subjects (18 to 60 years) with allergic
rhinoconjunctivitis asthma sensitised to
Phleum pratense pollen was carried out. A
total of 42 patients were enrolled. The
principal variable of the study was the
number and seriousness of both local and
systemic adverse reactions. Secondary clini-
cal end points included immunoglobulin
determination: specic IgE, total IgG and
IgG4 and dose-response prick test. Three
different build up phase schedules were
assayed, one involving six injections on a
weekly basis (A), another one of eight
injections (B) and a clustered administra-
tion of four double injections (C). Each
group included 14 patients. Concentrations
ranged from 10 to 1.000 TSU/ml.
Result: Group A administrations gave rise
to 26 local reactions and 4 systemic reac-
tions (1 graded I and 3 graded II). Group
B showed 35 local and 8 systemic (2
graded I and 1 graded II) reactions.
Finally, group C patients experienced 20
local and 9 (5 graded I and 4 graded II)
systemic reactions. None of the local
events recorded were considered large reac-
tions. Following injections with Timothy
pollen extracts allergen specic IgE, IgG
and IgG4 increased and skin reactivity to
P. pratense was reduced. All schedules
allowed reaching the dose of 500 TSU per
injection that could be established as maxi-
mum tolerated dose. None of the patients
discontinued treatment throughout the
study.
Conclusion: SCIT vaccines containing P.
pratense depot extracts have shown a very
good safety and tolerability prole and
efcacy in terms of humoral and dose-
response prick test. Among the administra-
tion schedules used, Group A has shown a
better tolerability and efcacy prole com-
pared to Groups B and C.
Allergy 66, Suppl. 94 (2011): 482642  2011 John Wiley & Sons A/S
1323
Intralymphatic allergen-specific immuno-
therapy is effective and safe for treat-
ment of birch pollen-induced allergic
rhinitis
Petterson, T1; Latif, L2; Mansson, A1; Westin, U2;
Cardell, L1
1
Karolinska Institutet, Div. of ENT diseases, CLINTEC,
Karolinska University Hospital Huddinge, Stockholm,
Sweden; 2Skane University Hospital, Dep. of
otorhinolaryngology, Malmo, Malmo, Sweden
Background: Allergen-specic immunother-
apy (ASIT) is commonly used to treat
patients with IgE-mediated pollen-induced
allergic disorders. In addition to providing
good symptom control, it is the only treat-
ment that induces persistent tolerance
against allergens and affects the long-term
development of allergic rhinitis and
asthma. The dominating route for adminis-
tration is subcutaneous and although the
maintenance dose can be reached through
various regimes, the current treatment pro-
tocol takes several years to complete. How-
ever, emerging evidence suggests that direct
intralymphatic injections could yield a fas-
ter benecial result with a lower allergen
dose. In this study we have evaluated both
immunological and cellular effects of intra-
lymphatic immunotherapy.
Method: Patients with birch-pollen induced
allergic rhinitis were recruited and treated
with three intralymphatic injections of
ALK Alutard (1,000 SQ-U Betula Verru-
cosa with an interval of 4 weeks) or pla-
cebo (0.9% NaCl) and asked to estimate
the pain associated with the injections.
Patients treated with conventional subcuta-
neous ASIT (maintenance dose 100,000
SQ-U given every 68 weeks during
3 years) were used as an additional control
group. Pre and post treatment, the patients
were evaluated by nasal allergen provoca-
tion, symptom scoring and skin prick test-
ing. Additionally, blood samples were
collected and analyzed for IgE, leukocyte
differentiation and various immunological
responses.
Result: All patients given intralymphatic
ASIT completed the treatment without any
severe adverse events and reported that the
injections were virtually painless. Prelimin-
ical situation appeared denitely improved.
ary data suggest a reduction of allergen-
specic IgE, a decrease in symptom scores
after allergen challenge along with altera-
tions in leukocyte phenotypes after com-
pleted treatment. The results appear to
mirror the effects seen with subcutaneous
ASIT and the patients receiving placebo
did not show any improvements.
Conclusion: Based on this limited study
population, it appears as intralymphatic
ASIT is safe for the patients and that
direct intralymphatic injections with a
1000-fold lower allergen dose than conven-
tional subcutaneous ASIT could yield simi-
lar benecial effects.
1324
Case report: successful nickel desensiti-
sation in a woman affected by contact
dermatitis
Bruno, M1; Campolo, Q2
1
Lofarma S.p.A., Medical Department, Milan, Italy;
2
Heath Clinic Dr. Lamberti-Castronuovo, Allergology
and Clinical Immunology Department, Reggio Calabria,
Italy
Background: About a year and a half ago,
a woman 28 years old, working as a
beauty advisor in a perfumery from
3 years, presented a severe hand dermatitis.
She had consulted many specialists and
tried various drug therapies without any
appreciable result. The patient reported an
improvement of the disease in the summer
season.
Method: The patient presents a clinical pic-
ture, characterized by redness and intense
itching at both hands with desquamation
and bleeding skin lesions, more evident on
the right hand. After the interruption of the
drug therapy for 10 days, a Sidapa series
patch test was performed: it was read 72 h
later showing a positivity to nickel
sulphate. An oral drug therapy with predni-
sone, ranitidine, hydroxyzine and lactoba-
cilli was prescribed in addition to
hydrocortisone butyrate ointment and bar-
rier cream, cleansing oil for daily hygiene
and white cotton gloves for skin protection.
As the patient reported gastro-enteric
symptoms, a low nickel diet was prescribed
as well. In the meantime, an oral hyposensi-
tization treatment to nickel was prescribed.
Result: After 20 days the patient returned
for check-up. As the clinical picture
appeared improved, the drug therapy was
revised maintaining lactobacilli, cleansing
oil and barrier cream for the hands, gloves,
low nickel diet and hydroxyzine. The hypo-
sensitization treatment has been performed
according to the scheme indicated by the
manufacturing company (Lofarma S.p.A.,
Milan, Italy). Eight months after the onset
of the hyposensitization treatment, the clin-
The hand skin has returned to normality.
The patient had to use the other therapies
only in few occasion. The gastro-enteric
symptoms were improved. Nowadays, after
a year and a half from the beginning of
the hyposensitization therapy, the patient
presents no lesion on the hands and only
uses cleansing oil and barrier cream. She
still selects the intake of foods containing
nickel but does not present gastro-enteric
symptoms any more.
Conclusion: Already after 3 months oral
hyposensitization treatment, the patient
503

showed an improvement of the clinical
symptomatology; such improvement has
become progressive and durable through-
out time. Many other patients have been
previously treated with this hyposensitiza-
tion treatment. However, the reported case
is intended to show that the use of the
hyposensitization therapy can solve also
severe cases.
1325
Allergic conjunctivitis and nickel
Bruno, M1; Cannao, G2; Russo, A2; Cannao, P3
1
Lofarma S.p.A., Medical Department, Milan, Italy;
2
ARNAS GARIBALDI, U.O.C. Internal Medicine,
Catania, Italy; 3General Hospital, University, Eye Clinic,
Messina, Italy
Background: Nickel is the main cause of
professional and extraprofessional allergic
dermatitis. One of the manifestations of
the disease are the conjunctival symptoms.
Aim of this study was to evaluate the fre-
quency of positivity to nickel in patients
with a positive history for conjunctival
symptoms.
Method: The patch test method with posi-
tivity to nickel sulphate, read after 72 h
(Lofarma S.p.A., Milan, Italy) has been
used in 15 patients (9 women and 6 men,
mean age 37 years). They were positive to
skin-prick test as well: 3 to Parietaria
Ofcinalis, 3 to Parietaria Judaica, 3 to
Dermatophagodes Ptneronyssinus, 1 to
Dermatophagodes Farinae, 1 to latex, 1 to
Grass-Mix. As a control group, 12 patients
with allergic conjunctivitis, however with a
negative patch test for nickel sulphate have
been used.
Result: The patch test with nickel sulphate
was able to identify all the subjects with
the disease due to nickel and to exclude all
those with the disease but without a sensi-
tization to nickel.
Conclusion: The results of the study dem-
onstrate the advantages of performing a
patch-test with nickel sulphate in patients
suffering from allergic conjunctivitis to be
addressed to specic immunotherapy with
oral nickel hyposensitizing treatment.
1326
Epicutaneous immunotherapy requires
intact skin and not stripped skin to prop-
erly activate and mature dendritic cells
toward induction of desensitisation
Dioszeghy, V1; Mondoulet, L1; Dhelft, V1; Ligouis, M1;
Puteaux, E1; Dupont, C2; Benhamou, P1
1
DBV-Technologies, Research and Development, Paris,
France; 2Universite Paris Descartes Hopital Necker,
Paris, France
Background: Epicutaneous immunotherapy
(EPIT) has been described in humans and
504
Poster Session 53 Novel clinical and basic aspects of allergen immunotherapy
animal models. We have shown that pro-
longed and repeated skin application of an
allergen (using the Viaskin (V) delivery
system), improves tolerance in sensitized
patients and mice, contrasting with the pre-
viously described sensitizing effect of appli-
cations of allergen on stripped skin in
mice. The aim of this study was to identify
the mechanisms involved in these opposite
situations.
Method: In BALB/c mice sensitized to
OVA with alum, EPIT was carried out
with V applied on intact or tape-stripped
skin, one time or repeatedly once a week
for 9 weeks. V coated with Alexa488-con-
jugated OVA (OVA*) was applied to ana-
lyze the phenotype of migrating dendritic
cells (DC) and lymph node (LN) cell
responses. After 9 weeks, effect of EPITs
on OVA-specic cytokine responses and
regulatory T cell (Treg) in spleen were
compared to Sham treatment.
Result: On intact skin, after a unique appli-
cation of V-OVA*, migrating DCs
expressed a high level of MHC-II,
62.2 3.70% of DCs expressing high lev-
els of CD205, CD86, and CD83. Migrating
DCs also displayed high levels of the inhibi-
tory receptor PIR-B and were positive for
intracellular IL-12 (iIL12). This was associ-
ated with a mixed response of LN cells with
OVA-specic production of IL5 (96.3
26.8 pg/ml), IL10 (138.7 24.9 pg/ml)
and IFNc (91.6 4.6 pg/ml). On stripped
skin, by contrast, after a unique application
of V-OVA*, only 42.7 3.49% of
migrating DCs were CD205high and DCs
expressed lower levels of MHC-II, PIR-B
and no iIL12. Stripping of skin also skewed
the LN response toward a Th2 prole with
signicantly higher production of IL5 and
lower production of IL10 and IFNc
(respectively 342.5 133.2 pg/ml; 75.2
12.1; 49.6 11.4 pg/ml; P < 0.05 versus
intact skin). On intact skin, after 9 applica-
tions of V, migrating DCs were activated
and matured comparably to the unique
application. OVA-specic cytokine responses
in spleen were down regulated and associ-
ated with more Treg, compared to Sham
(P < 0.05). On stripped skin, by contrast,
after 9 applications of V, migrating DCs
were CD205 low and expressed low level of
MHC-II, CD86, PIR-B and iIL12. Strip-
ping during EPIT prevented the decrease of
OVA-specic responses in spleen and the
induction of Treg.
Conclusion: EPIT on intact skin allows the
delivery of allergen which induces specic
activation and maturation of migrating
DCs, leading to desensitization via a Treg-
dependant mechanism.
Allergy 66, Suppl. 94 (2011): 482642  2011 John Wiley & Sons A/S
1327
IL-4, IL-10, TNF-a and IL-1a serum levels
in asthma patients may predict those
who are responders or non-responders
to allergen specific immunotherapy
Beloglazov, V1; Znamenska, L1; Bisyuk, Y1; DuBuske, L2
1
Crimean State Medical University, Simferopol,
Ukraine; 2Immunology Research Institute of New
England, Gardner, MA, United States
Background: The efcacy of immunother-
apy in asthma appears to be related to
both asthma severity and duration, indicat-
ing that the intensity of resulting inamma-
tory processes may be a key factor for
allergen specic immunotherapy (ASIT)
being a success or failure.
Method: A total of 49 patients with atopic
asthma with sensitization to ragweed and
having GINA Step I or II disease received
pre-seasonal ASIT. Depending on the effec-
tiveness of ASIT (during the peak of the
ragweed season), patients were divided into
2 groups: Group 1 39 (responders to ASIT
average total score in the period of aller-
gen pollination by ACQ questionnaire was
0.75 and below) and Group 2 10 patients
(non-responders to ASIT average total
score by ACQ was above 0.75). The control
group included 30 volunteers who were
blood donors of similar age and gender. IL-
4, IL-10, TNF-a and IL-1a in blood were
measured by ELISA methods before and
after ASIT.
Result: Group 1 and Group 2 had IL-4 that
was increased before ASIT (Group 1
38.32 5.52 pg/ml, Group 2 45.29
3.44 pg/ml) compared to the control group
(18.88 6.36 pg/ml, P < 0.01) which
decreased after ASIT in Group 1 (25.41
5.42 pg/ml, P = 0.04) only; IL-10 before
ASIT (Group 1 14.87 0.82 pg/ml,
Group 2 13.89 2.13 pg/ml) and after
ASIT (Group 1 14.49 1.47 pg/ml,
Group 2 13.95 2.20 pg/ml) was not sig-
nicantly different from the control group
(11.91 1.88 pg/ml, P > 0.05); TNF-a
before ASIT (Group 1 2.30 0.20 pg/ml,
Group 2 2.40 0.26 pg/ml) and after
ASIT (Group 1 2.20 0.21 pg/ml, Group
2 2.75 0.37 pg/ml) was signicantly
greater compared to the control group
(1.23 0.12 pg/ml, P < 0.01) and did not
change during ASIT; IL-1a did not change
in Group 1 during ASIT (before - 6.50
2.53 pg/ml, after 7.50 2.08 pg/ml) com-
pared to the control group (6.65 1.02 pg/
ml, P > 0.05) and was greater in Group 2
(before ASIT 12.15 1.29 pg/ml, after
ASIT 17.20 0.37 pg/ml, P < 0.05).
Conclusion: Low efcacy of ASIT in atopic
asthma may be associated with increased
levels of IL-4 and IL-1a, suggesting that
serum markers of inammatory responses
may assist in predicting efcacy of ASIT in
atopic asthmatics.
Poster Session 53 Novel clinical and basic aspects of allergen immunotherapy
1328
Epicutaneous immunotherapy in a mice
model of milk allergy
Mondoulet, L1; Dioszeghy, V1; Puteaux, E1; Ligouis, M1;
Dhelft, V1; Dupont, C2; Benhamou, P1
1
DBV Technologies, Paris, France; 2Necker Hospital,
Paris, France
Background: Epicutaneous immunotherapy
(EPIT) has been recently described as an
efcient and safe non invasive method of
immunotherapy in a mouse model of pea-
nut allergy. This study evaluated the ef-
cacy of EPIT in a mice model of milk
allergy, particularly focusing on the diges-
tive tract injuries after exclusive milk diet.
Method: After oral sensitization with milk
and cholera toxin, 10 BALB/c mice were
treated weekly during 8 weeks by pow-
dered milk applied on intact skin using an
epicutaneous delivery system (EPIT), and
were compared to 10 Sham (sensitized, non
treated) and 10 controls. After EPIT, all
animals received an exclusive milk diet for
10 days then were sacriced under general
anaesthesia. Specic (s) IgE, IgG1 and
IgG2a and spleen cell responses were mon-
itored. Esophageal and jejunal samples
were taken for histology and mRNA
expression of cytokines.
Result: sIgE increased after oral sensitiza-
tion, in EPIT and Sham (respectively 0.13
and 0.14 lg/ml). After treatment, sIgE
increased in Sham, 0.32 versus 0.14 lg/ml
(P < 0.01) in EPIT, whereas sIgG2a
increased only in EPIT, 0.17 versus
0.06 lg/ml (P < 0.05) in Sham. Th2 cyto-
kines secreted by reactivated splenocytes
decreased with EPIT compared to Sham:
IL-4 (367 versus 975 pg/ml, P < 0.05), IL-
5 (1112 versus 2742 pg/ml, P < 0.05), IL-
10 (5340 versus 9965, ns) and IL-13 (4166
versus 7065, P < 0.05). After exclusive
milk feeding, mRNA expression of eotaxin
and IL-5 increased in Sham compared to
EPIT: eotaxin (1.26 versus 1.10, ns) and
IL-5 (1.61 versus 1.13, P < 0.05). Esopha-
geal eosinophilic inltration (measured in
six high power elds) was higher in Sham,
43 cells/mm2, than in EPIT, 19 cells/mm2
(P < 0.05) and naive, 11 cells/mm2
(P < 0.01). Esophageal mucosa thickness
was considerably increased in Sham
(P < 0.01) versus EPIT, the latter being
comparable to naive.
Conclusion: In sensitized mice, EPIT pre-
vents allergy symptoms triggered by oral
milk consumption, especially the esopha-
geal digestive lesions.
Allergy 66, Suppl. 94 (2011): 482642  2011 John Wiley & Sons A/S
1329
Epicutaneous versus sublingual and sub-
cutaneous immunotherapy in a model of
peanut sensitised mice: a doseeffect
study
Mondoulet, L1; Dioszeghy, V1; Ligouis, M1; Dhelft, V1;
Puteaux, E1; Dupont, C2; Benhamou, P1
1
DBV Technologies, Paris, France; 2Necker Hospital,
Paris, France
Background: The aim of this study was to
compare the efcacy of three different
methods of immunotherapy in a dose
effect study with peanut-sensitized mice.
Method: After oral sensitization with pea-
nut and cholera toxin (n = 100 mice),
BALB/c mice were treated by epicutaneous
(EPIT), sublingual (SLIT) or subcutaneous
(SCIT) routes using three dosages (20, 100,
500 lg) and compared to 10 mice sensi-
tized but not treated (Sham) and 10 naive
mice. After treatment, spleens were har-
vested and cells reactivated in vitro in order
to measure the prole of secreted cyto-
kines. Specic antibodies were monitored
during immunotherapy.
Result: EPIT induced a signicant specic
(s) IgE decrease with a treatment dosage of
100 lg (from 0.22 0.03 to 0.09
0.01 lg/ml, P < 0.05) and 500 lg (from
0.28 0.03 to 0.09 0.02 lg/ml,
P <0.05). SCIT induced a decrease with
100 lg (from 0.18 0.05 to 0.09
0.01 lg/ml, P < 0.05) but not with 500 lg.
SLIT did not modify sIgE levels. The 20 lg
dosage did not induce any sIgE modication
whatever the route. sIgG2a exhibited a sig-
nicant increase with EPIT 20 lg (from
0.49 0.12 to 6.67 3.07 lg/ml,
P <0.05), EPIT 100 lg (from
0.05 0.0.01 to 5.03 2.30 lg/ml,
P < 0.05), EPIT 500 lg (from 0.44 0.29
to 4.38 1.36 lg/ml, P < 0.05) and SCIT
100 lg (from 1.9 0.23 to 7.04 2.86 lg/
ml, P < 0.05). No modication was
obtained with any other dosages or SLIT
groups. Compared to Sham, IL-4 (377 pg/
ml), IL-5 (1421 pg/ml) and IL-13 (4219 pg/
ml) decreased signicantly with EPIT
100 lg (225 pg/ml, 750 pg/ml, 2055 pg/ml,
P <0.05), EPIT 500 lg (192 pg/ml, 960 pg/
ml, 2180 pg/ml, P < 0.05), SLIT 100 lg
(172 pg/ml, 678 pg/ml, 1657 pg/ml,
P <0.05) and SCIT 100 lg (139 pg/ml,
774 pg/ml, 997 pg/ml, P < 0.05). No modi-
cation was induced with the other dosages.
Conclusion: EPIT, SCIT and SLIT had the
same efcacy at a dose of 100 lg. The
lower dosage (20 lg) had no therapeutic
effect and the higher dosage (500 lg) acted
only with EPIT.
1330
Recent trends in treatment strategies
and the usage of different probiotics bac-
teria
Alaeddini, B1; Roozbeh Nasiraie, L1; Basirnezhad, M2;
ketabdar, H3; Falak, R2; Tehrani, M4; Varasteh, A2
1
Department of Food Science, Nour Branch, Islamic
Azad University, Nour, Islamic Republic of Iran;
2
Immunobiochemistry Lab, Immunology Research
Center, School of Medicine, Mashhad University of
Medical Sciences, Mashhad, Islamic Republic of Iran;
3
Department of biology science, Islamic Azad
University of Mashhad Branch, Mashhad, Islamic
Republic of Iran; 4Faculty of Medicine, Department of
Microbiology and Immunology, Mazandaran University
of Medical Sciences, Sari, Islamic Republic of Iran
Background: Purpose of review: Incidence
of allergic disorders have been increased in
western and industrial countries ve decades
late, that is associated with a lower preva-
lence of infectious diseases and limited expo-
sure to microorganisms because of more
hygienic lifestyle. In this context, the use of
probiotics bacteria appears as a valuable and
attractive approach for treatment of allergic
disorders. In this review, different treatment
strategies with probiotics will be discussed.
Method: It is a review paper.
Result: The most recent study focus on the
effects of probiotics in enhances and
matures immunosystem in infants and
induces immunomodulation. In this regard,
probiotics are recommended for prevention
and curing allergy diseases especially for
infants that suspected cows milk allergy.
Probiotics may serve as both attractive
adjuvant systems for improved allergy
treatment and as a new treatment
approaches that use selected probiotic
strains as mucosal antigen delivery vehicles
for recombinant allergens. Important
insights have been made, indicating that
the benecial effects of probiotics are
strain dependent, as well as dependent on
the mode and timing of treatment.
Conclusion: Further information about
mechanism of probiotics in treatment
allergy, especially useful strain and dose of
probiotics for administrating and transla-
tion of studies in animal models into clini-
cal trials, gives hope that probiotics in
different strategies can be a successful
treatment for allergy in humans.
1331
Efficacy of sublingual specific immuno-
therapy drops in patients with respira-
tory allergy to Alternaria alternata: a
randomised, assessor-blinded, patient-
reported outcome, controlled 3-year trial
Milani, M1; Pozzan, M2
1
ALK, Medical Department, Lainate, Italy; 2Servizio di
Allergologia, ASL 16 Distretto 1 Padova, Padova, Italy
Background: Alternaria alternata (AA) sen-
sitisation is a common cause of respiratory
505

allergies such as rhinitis and asthma. So
far there are no double-blind randomised
controlled trials evaluating the efcacy and
safety of specic sublingual immunother-
apy (SLIT) drops in AA allergies.
Objective: To evaluate the efcacy of SLIT
treatment in terms of clinical improvement
and symptomatic medication usage in
patients with conrmed AA respiratory
allergy.
Subjects and methods: A randomised, open
parallel group study in 52 subjects was
conducted (32 men, mean age
20 9 years) with clinically and labora-
tory conrmed AA allergic rhinoconjuncti-
vitis with or without mild to moderate
asthma. Patients were randomly assigned
to SLIT treatment (SLITone Alternaria,
ALK, Denmark) one monodose ampoule
per day without up-dosing for 3 consecu-
tive years (n = 34) or to a control group
(n = 18) with a random allocation ratio of
2:1. Clinical rhinoconjunctivitis symptoms
(runny nose, blocked nose, sneezing and
itchy nose; eye symptoms included gritty
feeling/red/itchy eyes and watery eyes)
improvement was assessed by evaluating
patient-reported outcome (PRO) using a
6 cm visual analogue scale (VAS) (0:
extreme worsening; 3: no change; 6:
extreme improvement). The VAS score was
evaluated every year with an intra- and
inter-group comparison. Symptomatic
medication score (MS) was assessed by
evaluating drug consumption.
Results: After 3 years the VAS score was
4.7 0.8 in the SLIT group and 2 1.6
in the control group (P < 0.0002).Clinical
improvement was observed in 33 out of 34
subjects in the SLIT group (97%) and in 5
out of 18 in the control group (27%)
(P < 0.0001; Fishers exact test). The MS
signicantly (P = 0.0001) decreased in the
SLIT group from 4.3 to 1.7 at the end of
3 years treatment. In the control group
MS increased from 3.4 to 4.0 by the end of
the trial. No serious adverse events were
observed in the either group. Six patients
in the SLIT group (17%) reported side-
effects, in general mild and transient in
nature. One patient prematurely discontin-
ued the treatment due to gastrointestinal
discomfort.
Conclusion: The current study has shown
that a 3-year course of SLIT is efcacious
and well-tolerated in subjects with AA
induced respiratory allergies.
506
Poster Session 53 Novel clinical and basic aspects of allergen immunotherapy
1332
Allergen-specific immunotherapy with
complexes of modified allergens and
polymeric nanoparticles
Babakhin, A1; Maksimenko, O2; Gelperina, S2; Andreev,
S1; Khaitov, M1; DuBuske, L3
1
Institute of Immunology, National Research Center,
Moscow, Russian Federation; 2Nanosystem Ltd.,
Moscow, Russian Federation; 3Immunology Research
Institute of New England, Gardner, MA, United States
Background: This study investigates modu-
lation of allergic antibody responses with a
modied allergen adsorbed on polymeric
nanoparticles (NP).
Method: Polylactide-coglycolide (PLGA)
nanoparticles with end COOH groups
(PLGA-COOH) were synthesized (size:
175 46 nm). Basophil histamine release
was detected by the glass-ber based hista-
mine release assay. Ovalbumin (OVA) was
succinylated (sOVA) with modication of
80% of epsilon-amino groups. Complexes
of sOVA and PLGA-COOH NPs were pre-
pared. (CBAxC57Bl/6)F1 mice were immu-
nized three times with 10 lg OVA/mouse.
Between the second and third immuniza-
tions, treatment groups included:(1) 12 s/c
injections of saline (controls); (2) 12 s/c
injections of OVA (increasing doses from 1
to 1000 lg/mouse); (3) 6 s/c injections of
sOVA (increasing doses from 100 to
1000 lg/mouse); (4) 4 s/c injections of
sOVA and NPs (two doses of 100 lg and
two doses of 1000 lg sOVA/mouse; NPs
2 mg/mouse); (5) 4 s/c injections of PLGA-
COOH NPs only (2 mg/mouse); and (6)
non-immunized and non-treated mice (con-
trols). Serum levels of anti-OVA IgE, IgG,
IgG1, IgG2a antibodies (Abs) before, dur-
ing, and after ASIT were detected by
ELISA.
Result: NPs did not induce basophil hista-
mine release from atopic and non-atopic
donors. Group 4 induced anti-OVA IgE
Abs after the second and the third immuni-
zations similar to that of Group 3 but
slightly greater than Groups 1 and 2. Anti-
OVA IgG Abs in Groups 2, 3, and 4 were
approximately at the same level and
greater than Group 1. Anti-OVA IgG1
Abs in Groups 2, 3, and 4 were greater
than Group 1 but anti-OVA IgG2a Abs
were similar in Groups 1 through 5.
Conclusion: Complexes of modied aller-
gen and polymeric NPs induced strong
anti-allergen IgG responses suggesting the
use of NPs as adjuvants for the develop-
ment of novel vaccines for ASIT.
Allergy 66, Suppl. 94 (2011): 482642  2011 John Wiley & Sons A/S
1333
Possibility of immunotropic therapy
application in patients with acne vulgaris
Pavlenko, T1; Sizjakina, L1; Sidorenko, O2
1
Rostov-on-Don State Medical University, Clinical
Immunology and allergology department, Rostov-on-
Don, Russian Federation; 2Rostov-on-Don State Medical
University, Dermatology and Venerology department,
Rostov-on-Don, Russian Federation
Background: Acne vulgaris is the most
common skin pathology. Acneform disease
is accompanied by secondary immune de-
ciency, which is associated with intensive
growth of pathogenic microora bacteria
and different fungi. Standard therapy is
not always effective and using of systemic
and external immunotropic medications in
complex of acne vulgaris treatment is
debating. The aim of this investigation is
studying the efciency of immunotropic
medications in acne vulgaris treatment.
Methods: 36 patients at the age 16
25 years with moderate and severe acne
vulgaris, complicated with scarring, partici-
pated in the research. While investigating
the immune system there was determined
the content of CD3+, CD4+, CD8+,
CD16+, CD20+, CD25+, HLA-DR,
D95+ lymphocytes; dened immune
serum globulin A, M, G classes; studied
the oxidationreduction process rates of
neutrophils in Nitro Blue Tetrazolium
Reduction Test; level of circulating
immune complexes. The patients were
divided in two equal groups the control
group and the experimental one. The
patients of the standard group had com-
mon therapy-cream, containing 0.1%
adapalene. The experimental group
patients had therapy, consisting of complex
application of standard therapy of acne-
form disease and a course of ozone therapy
in the form of local subcutaneous injec-
tions of ozoneoxygen mixture with con-
centration 3500 mcg/ml. The ozone
procedures were performed twice a week,
total of 10 procedures. In addition to stan-
dard external therapy, the experimental
group patients were treated with cream,
containing epigallocatechin-3-gallate. The
total duration of treatment was 1 month.
Results: The analysis of parameters for
immune changes in the experimental group
has shown a statistically reliable increase
of CD4+ lymphocytes (from 32.2 1.3%
to 41.9 1.9%), which results in improve-
ment of the helpersuppressor cell ratio
values, rise of serum IgG (from
10.4 0.35% to 13.2 1.4%), improve-
ment of elimination functions which is ver-
ied by decreasing in quantity of the
circulating immune complexes (from
90.1 2.4 to 64.9 1.9).
Conclusion: Using of combination of
ozone-therapy and external medicine,
Poster Session 53 Novel clinical and basic aspects of allergen immunotherapy
containing epigallocatechin-3-gallate with
standard methods in patients with moder-
ate and severe acne vulgaris with tendency
to scarring results in signicant improve-
ment of clinical and immunological values.
1334
Patient management in the sublingual
immunotherapy of house dust mite respi-
ratory allergies
Trebuchon, F1; David, M2; Demoly, P3; Robin, B4
1 the patients were treatment-compliant.
France; 2Stallergenes SA, Antony, France; 3University
Hospital of Montpellier, Montpellier, France;
4
Stallergenes, Antony, France
Background: Little information is available
on how specialist physicians apply sublin-
gual immunotherapy (SLIT) to treat house
dust mite (HDM)-induced allergic respira-
tory diseases.
Method: In this observational, retrospec-
tive, multicentre study in France, 139 spe-
cialists recruited 1289 patients of all ages
in two groups (those seen in consultation
in 2002 and 2005). The physicians lled
out a case report form covering clinical
aspects, health-related quality of life and
treatment modalities.
Result: The populations mean SD age
was 20.50 14.34. Most of the patients
suffered from rhinitis with asthma (52.7%
and 49.5% in 2002 and 2005, respectively).
Rhinitis alone was present in 44.4% and
46.7% of the patients, respectively. The
patients generally suffered from moderate-
to-severe rhinitis (70% and 76%, respec-
tively). Overall, 85% of the patients
considered that their symptoms (rhinor-
rhoea, sneezing, nasal obstruction, nasal
pruritus) were troublesome. Following
SLIT initiation, a signicant drop in the
proportion of patients on oral antihista-
Allergy 66, Suppl. 94 (2011): 482642  2011 John Wiley & Sons A/S
mines and nasal corticosteroids was
observed (38% and 21.6%, respectively,
for patients suffering from rhinitis with or
without asthma). For asthma with or with-
out rhinitis, the physicians observed
decreases of 32%, 25% and 16% in the
number of patients on antihistamines,
inhaled corticoids and b2 agonists, respec-
tively. Improvements in asthma and rhini-
tis were evidenced in 59.1% and 61.1% of
the patients. The SLITs safety prole was
good (with mainly local reactions). 84% of
76.6% of the patients considered their
SLIT to be effective or very effective and
78.4% were satised with their treatment.
Conclusion: Patients with moderate or
severe HDM-induced allergic respiratory
diseases noted an improvement in their
symptoms during SLIT. A signicant
reduction in concurrent use of symptom-
atic medications was notably observed.
Most of the patients were satised with the
SLIT prescribed by their physician.
1335
Clinical efficacy of sublingual immuno-
therapy in seasonal allergic asthma
Stosovic, R; Bogic, M; Peric Popadic, A; Tomic Spiric,
V; Bolpacic, J; Raskovic, S; Savic, N; Stefanovic, L;
Djuric, V
Clinical Center of Serbia, Institute of Allergology and
Immunology, Belgrade, Serbia
Background: The evaluate of efcacy of
sublingual allergen specic immunotherapy
(SLIT) in seasonal allergic asthma (SAA)
have been performed predominantly by
short-term studies in monosensitised
patients. The aim of the study was to
investigate short-term clinical effects of
SLIT added to treatment with medical
therapy (MT) in comparation to MT only
in patients with SAA polisensitised to grass
and ragweed pollen.
Method: The prospective (randomized and
controlled) study covered 17 adolescent
and adult patients with mild to moderate
severe SAA (with or without seasonal aller-
gic rhinitis) induced by hypersensitivity to
grass and ragweed pollen, indicated to
SLIT. The experimental group was com-
posed of 10 patients subjected to precosea-
sonal SLIT (antiallergic biologically
standardized vaccines, sublingual drops)
and MT, while seven adequate matched
patients on MT only served as controls.
The clinical efcacy of SLIT was evaluated
by the symptom-drug (diary card) score
(SDS) measurement, skin prick test (SPT)
and non-specic bronchial hyperactivity
(NBH) measurement (nonspecic bronchial
provocation test by metacholine), before
initiation of SLIT and 1 year later (in pol-
len season and after pollen season).
Results: The SDS were signicantly
reduced in patients subjected to SLIT
(P < 0.01) 1 year after the onset it. SPT
to grass and ragweed pollen also was sig-
nicantly reduced (P < 0.05) but NBH
did not change. In patients receiving MT
only, SDS was also signicantly reduced
(P < 0.01), but signicantly higher
(P < 0.05) from the value obtained in the
experimental group. SPT and NBH
remained unchanged and signicantly
higher (P < 0.05) then in experimental
group.
Conclusion: SLIT added to MT is clinically
effective in patients with seasonal allergic
asthma polisensitised to grass and ragweed
pollen.
507

Immunological advances in allergen immunotherapy
1336
Pharmacokinetics of a major Japanese
cedar allergen, Cry j 1, for subcutaneous
or sublingual immunotherapy in the rat
Ohashi-Doi, K1; Kashiwabara, S1; Oka, H1; Mitobe, Y1;
Ikeda, S1; Takao, A2; Kashiwagi, T1
1
TORII Pharmaceutical CO. LTD, Research Laboratory,
Sakura, Japan; 2SEKISUI Medical Co Ltd, Research
Institute, Ibaraki, Japan
Background: Specic allergen immunother-
apy is an effective treatment for rhinitis
and asthma caused by allergic sensitization.
Sublingual immunotherapy (SLIT) is clini-
cally effective, and was introduced as an
alternative to subcutaneous immunother-
apy (SCIT) to solve the problem of sys-
temic reaction. Presently, SLIT has been
developed into represent an advanced clini-
cal application. However, little is known
about the pharmacokinetics of allergens
for local immunotherapy. Therefore, this
study aimed to assess the pharmacokinetics
of a major Japanese cedar allergen, Cry j
1, in rats following SLIT or SCIT.
Method: Radiolabeled (125I)-Cry j 1 was
synthesized. 125I-Cry j 1 had a specic
activity of 1.48 MBq/lg and radiochemical
purity of >98%. Male Sprague-Dawley
rats (7-week-old) were used. The radiola-
beled 125I-Cry j 1 (7.5 lg/rat) was adminis-
tered by either SCIT (50 ll/rat) or SLIT
(5 ll/rat). The approximate radioactivity
dose was 0.440.50 MBq. To measure the
radioactivity, blood samples were collected.
The tissue radioactivity was measured at
0.5, 2 or 4 h after administration. All pro-
cedures employed in the experiments were
approved by the Animal Ethics Committee
in accordance with the Laboratory Animal
Welfare guidelines.
Result: In the SCIT-treated rats, the radio-
activity was widely distributed in all tissues
except for the central nervous system. The
mean apparent Cmax of the total radioac-
tivity concentration in plasma was 31 ng-
eq. Cry j 1/ml. The Tmax in plasma was
achieved at 2-h post-administration. The
unchanged-Cry j concentration in plasma
was signicantly reduced in a time-depen-
dent manner in SCIT-treated rats. The
SLIT-treated rats showed a time-dependent
increase in the radioactivity concentration
in plasma. The unchanged Cry j 1 concen-
tration was approximately 0.100.49 ng/ml.
The tissue radioactivity analysis revealed
508
Poster Session 54
that the concentration was signicantly
higher in the mandibular lymph node than
in the plasma, and the tissue/plasma ratio
of the radioactivity concentration was 2.0
5.1 after 0.54 h in the SLIT-treated rats.
Conclusion: This is the rst report of the
distribution of a major Japanese cedar
allergen, Cry j 1, in rats. The results
showed that Cry j 1 was less likely to be
absorbed into the blood after SLIT than
after SCIT in rats. The radioactivity
detected in the mandibular lymph node
after SLIT suggests that the mandibular
lymph node may play a key role in the
mechanism of action of SLIT.
1337
Subcutanous sensitisation with high anti-
gen doses without the use of adjuvant
prevents the induction of IgE production
and airway hyperresponsiveness in
experimental allergic airway inflamma-
tion
Raker, V1; Maxeiner, J2; Montermann, E1; Taube, C2;
Grabbe, S3; Reske-Kunz, A1; Sudowe, S1
1
Department of Dermatology, Clinical Research Unit
Allergology, University Medical Center of the Johannes
Gutenberg University, Mainz, Germany; 2University
Medical Center of the Johannes Gutenberg University
Mainz, III. Medical Clinic, Asthma Core Facility, Mainz,
Germany; 3Department of Dermatology, University
Medical Center of the Johannes Gutenberg University
Mainz, Mainz, Germany
Background: The application of high aller-
gen doses during specic immunotherapy
leads to allergen-specic tolerance includ-
ing the inhibition of IgE production.
Recently, we identied CD4-CD8-double-
negative T cells as potent suppressor cells
for antigen-specic IgE immune responses
arising after repetitive intraperitoneal injec-
tion of high doses of keyhole limpet hemo-
ycyanin (KLH) adsorbed to the adjuvant
aluminiumhydroxide. This work focuses on
an alternative immunization protocol,
which favours subcutaneous, adjuvant free
application of KLH or ovalbumin (OVA)
as clinically relevant allergen.
Objective: To analyze antigen dosedepen-
dent immune responses after repeated sub-
cutaneous sensitization without adjuvant in
a mouse model of allergic airway inam-
mation.
Method: BALB/c or C57BL/6 mice were
immunized subcutaneously with low doses
(LD) or high doses (HD) of KLH or OVA
Allergy 66, Suppl. 94 (2011): 482642  2011 John Wiley & Sons A/S
as antigen. Content of antigen-specic IgE
in sera was determined after the sixth
immunization. Antigenic restimulation of
spleen cells was performed in vitro. At the
end of the sensitization phase mice were
challenged with antigen for 3 days by
intranasal instillation. Subsequently, airway
reactivity was measured after methacholine
challenge by invasive measurement and
lung histology was performed.
Result: In contrast to LD mice, which
developed a strong IgE immune response,
HD mice showed reduced IgE production
as well as symptoms of tolerance induction
indicated by reduced secretion of Th2 cyto-
kines (IL-5, IL-13) by and hypoprolifera-
tion of splenocytes after restimulation
in vitro. The suppression of IgE production
in HD mice was long-lasting and effective
and could not be reversed by subsequent
immunization with low antigen doses. The
antigen dosedependent regulation of IgE
production could be veried in TLR-4-/-
mice, implying that LPS contamination of
antigen is not responsible for the inhibitory
effects on IgE production observed in HD
mice. Whereas LD mice showed airway
hyperreactivity (AHR) after intranasal
challenge, HD mice did not develop AHR.
Furthermore, HD mice showed markedly
reduced mucus production by goblet cells
after local provocation of the airways as
compared with LD mice.
Conclusion: Subcutaneous application of
high antigen doses prevents IgE immune
responses as well as on the induction of
AHR and lung inammation following
local challenge with antigen.
1338
Characterisation and cross-reactivity of
the human T cell response to the major
bahia grass pollen allergen, Pas n 1
Etto, T1; Gardner, L1; de Boer, C1; OHehir, R2; Rolland,
J1
1
Department of Immunology, Monash University,
Melbourne, Australia; 2Department of Allergy,
Immunology and Respiratory Medicine, Alfred Hospital,
Melbourne, Australia
Background: Bahia grass (Paspalum nota-
tum) pollen is a major cause of allergic rhi-
nitis and asthma, especially late in the
pollen season. Allergen-specic immuno-
therapy is used to treat patients with grass
pollen allergy, but those with moderate to
Poster Session 54 Immunological advances in allergen immunotherapy
severe asthma are excluded due to risk of
severe reactions including anaphylaxis. To
develop a safer specic treatment for Bahia
grass pollen allergy, T cell-reactive but
non-IgE-binding allergen variants are
required. This treatment may also be suit-
able for allergy to related grasses. Our aim
was to identify dominant CD4+ T cell-
reactive peptides of the major Bahia grass
pollen allergen, Pas n 1, and examine
cross-reactivity of Pas n 1-specic T cells
to other common grass pollen allergens.
Method: Peripheral blood mononuclear
cells from 19 Bahia grass pollen-allergic
subjects were cultured with Pas n 1 to gen-
erate Pas n 1-specic T cell lines (TCLs).
TCLs were screened for proliferative and
cytokine response to individual overlapping
20-mer Pas n 1 peptides. Pas n 1 peptide-
specic T cell clones (TCCs) were gener-
ated by single cell sorting of CD4+ T cell
blasts (CD4+CFSElo lymphocytes) into
96-well plates. Following expansion, TCCs
were tested for cross-reactivity to Lol p 1
and Cyn d 1, the major allergens of Rye-
grass and Bermuda grass pollen, respec-
tively. IgE reactivity of Pas n 1 peptides
was tested by dot blotting.
Result: Pas n 1-specic CD4+ T cell lines
were generated from all Bahia grass pollen-
allergic patients in this study. Three Pas n 1
20-mers were identied as the dominant T
cell-reactive peptides of Pas n 1 in our patient
population; 78% patients reacted to one or
more of these peptides. Two of the dominant
peptides showed T cell cross-reactivity with
homologous Lol p 1 and Cyn d 1 peptides.
None of these dominant peptides showed
binding to patient serum IgE antibodies.
Conclusion: We have identied the domi-
nant T cell-reactive peptides of the major
Bahia grass pollen allergen, Pas n 1. Two
peptides showed T cell cross-reactivity with
other grasses but one was unique for Bahia
grass. These T cell-reactive but non-IgE-
reactive peptides will be suitable for devel-
opment of a safe vaccine to induce
specic immune tolerance in Bahia grass
and other grass pollen-allergic individuals,
including asthmatics.
1339
The evaluation of venom immunotherapy
efficiency with basophil activation test
and sting challenge
Erzen, R1; Andreja, P2; Mira, S2; Kosnik, M2; Korosec, P2
1 and Environmental Sciences, Deakin University,
2
Hospital Golnik, Allergology, Golnik, Slovenia;
Slovenia
Background: The role of basophil activa-
tion test (BAT) in evaluating the efciency
of treatment with venom immunotherapy
(VIT) in Hymenoptera venom allergic
patients is poorly understood.
Allergy 66, Suppl. 94 (2011): 482642  2011 John Wiley & Sons A/S
Method: Subjects with conrmed history
of anaphylaxis (Muller grade IIIV) after
Hymenoptera sting and treated with VIT
were included into this prospective, single-
blinded study during years 20042010.
Basophil CD63 activation was measured
before VIT and at the end of build-up
phase in year 2004, one year later in 2005
and after VIT was nished in year 2010.
After the last BAT measurement a sting
challenge test was performed.
Result: 22 patients were included into the
study, 11 with hypersensitivity to wasp and
11 with hypersensitivity to honeybee
venom. Only one patient had a systemic
hypersensitivity reaction (Muller grade II)
after the sting challenge with honeybee. In
a group of patients with negative sting
challenge BAT level did not change after
1 year of VIT. However, after 5 years of
VIT we found a signicant decrease in
basophil allergen specic sensitivity, repre-
sented by a marked drop of CD63
response at sub maximal 0.1 lg/ml of
venom concentration. On the other hand
there were no changes at the level of baso-
phil reactivity represented by the CD63
response at maximal 1 lg/ml of venom
concentration. In a patient with positive
sting challenge, and therefore failed VIT,
the basophil CD63 allergen specic respon-
siveness did not decrease, but even increase
after completed VIT.
Conclusion: These results suggest that mea-
surement of basophile responsiveness might
be a useful marker for following and moni-
toring the efcacy of VIT. Moreover, it
seems that changes in basophil responsive-
ness might be important for long term pro-
tection achieved by VIT.
1340
CD25+CD4+Tregs mediate the protection
from oral peanut-induced esophageal
lesions of sensitised mice treated by
epicutaneous immunotherapy
Dioszeghy, V1; Mondoulet, L1; Dhelt, V1; Ligouis, M1;
Puteaux, E1; Dupont, C2; Benhamou, P1
1
DBV-Technologies, Research and Development, Paris,
France; 2Universite Paris Descartes Hopital Necker,
Paris, France
Background: In mice sensitized to peanut,
sustained oral ingestion of peanut protein
extracts (oral PPE) induces eosinophilic
inltration of the esophageal mucosa com-
parable to eosinophilic esophagitis (EoE)
and villous atrophy in jejunum. When sen-
sitized mice are previously treated by epi-
cutaneous immunotherapy (EPIT), lesions
decrease dramatically, together with a
decrease of IL5, IL13 and eotaxin expres-
sion and an increase of FoxP3 expression
in the esophageal mucosa (Mondoulet,
ESPGHAN 2010). These ndings suggest a
role for T regulatory cells (Tregs) in EPIT.
The aim of this study was to conrm the
EPIT-induced Treg mechanism in the pre-
vention of EoE occurring in sensitized mice
following oral PPE.
Method: First, two groups of eight Balb/c
mice were orally sensitized by PPE with
cholera toxin during 6 weeks, then treated
by EPIT using a patch (Viaskin) with
100 lg of PPE 48 h once a week for
8 weeks. In one group, Tregs were blocked
by injection of anti-mouse CD25 antibody
(aCD25) once a week for 8 weeks. The
two groups (EPIT and EPIT+ aCD25)
were compared to sensitized untreated
(Sham) and nave groups after oral PPE.
Second, CD25+CD4+ Tregs were sorted
from spleen cells of EPIT and Sham trea-
ted mice and transferred to PPE-sensitized
mice which then received oral PPE. At the
end of experiments, esophageal samples
were taken for histology and mRNA anal-
ysis.
Result: As previously described, oral PPE
induced EoE in Sham treated mice and
increased eotaxin expression, which were
prevented in EPIT treated mice (respec-
tively 36.39 5.61 versus 13.30 4.07 eo-
sinophils per mm2 (E/mm2), and 2.11
0.29 versus 1.17 0.15 eotaxin relative
mRNA (rmRNA), P < 0.05). Also, the
esophageal expression of FoxP3 increased
in EPIT compared to Sham (respectively
2.04 0.49 versus 085 0.15 rmRNA,
P < 0.05). aCD25 reversed the effects of
EPIT in the esophagus (38.93 7.13 E/
mm2, 2.35 0.62 eotaxin rmRNA and
0.71 0.18 FoxP3 rmRNA; P < 0.05 ver-
sus EPIT). Transferring Tregs from EPIT
treated mice to PPE sensitized mice sub-
mitted to oral PPE prevented eosinophil
inltration and eotaxin expression, and
induced FoxP3 in esophagus whereas cell
transfer from Sham treated mice demon-
strated no effect.
Conclusion: Induction of Tregs by EPIT is
able to prevent EoE in mice sensitized to
peanut conrming the Treg mediated
mechanism of EPIT.
1341
Characterisation of a novel IL4 receptor
antagonist for allergy treatment
Ahmed, N1; Dhanapala, P1; Carr, S1; Suphioglu, C2
1
School of Life and Environmental Sciences, Deakin
University, Burwood, Victoria, Australia; 2School of Life
Geelong, Victoria, Australia
Background: Allergy is a signicant source
of nancial, physical and psychological
stress, not only on those immediately
affected, but the society as a whole.
Unlike pollen allergy, allergy to peanut
can result in life-threatening reactions
509

causing morbidity and high economic cost.
Therefore, safer and more effective
reagents are urgently needed for peanut
allergy treatment. The interaction of cyto-
kines IL4 and IL13 with IL4 receptor
(IL4R) is critical in the allergic response,
which results in the production of aller-
gen-specic IgE antibodies by B cells.
Therefore, down-regulation of this interac-
tion is a desirable target to identify novel
allergy therapeutics that down-regulate
IgE production, which forms the overall
aim of this study.
Method: We have employed the innovative
phage display technology to screen a ran-
dom peptide library with a biologically
active puried human IL4R to identify
potential peptide candidates that interacted
with the receptor. Once potential peptides
were identied, they were commercially syn-
thesized and used in a number of in vitro
immunoassays, to assess their capacity in
inhibiting the IL4 and IL13 interaction with
the IL4R.
Result: We have successfully used random
peptide phage display library to identify
M13 phage clones that demonstrated spe-
cic binding to IL4R. All of the 10 individ-
ual phage clones characterised had
identical peptide sequence motifs, which
was used to synthesize the corresponding
peptide for use in ELISA, which demon-
strated signicant binding to the IL4R.
The capacity of the synthetic peptide to
inhibit IL4 binding to IL4R was investi-
gated with inhibition ELISA, showing sig-
nicant inhibition. As both IL4 and IL13
interact with the IL4R, competition
between the peptide antagonist and the
cytokines IL4 and IL13 in binding to the
IL4R was assessed with competition
ELISA. We have successfully shown that
although the peptide antagonist signi-
cantly inhibited both cytokines IL4 and
IL13 in interacting with the IL4R, IL4 was
shown to be a better competitor than the
IL13, which correlates with ndings in the
literature.
Conclusion: We have successfully identied
a novel synthetic peptide antagonist
against IL4R, which effectively blocks IL4
and IL13 interaction with the IL4R in vi-
tro. Since IL4 and IL13 interaction with
IL4R is a common pathway for many
allergies, a prophylactic treatment can be
devised by blocking this interaction for
treatment of allergies.
510
Poster Session 54 Immunological advances in allergen immunotherapy
1342
Conclusion: The modied Dp and Df aller-
Physicochemical characterisation of the gen preparations were well characterised
modification of a mite allergen extract by the applied physicochemical techniques.
Identication of the relevant allergens and
Luykx, D1; de Bruijn, J1; Sleijster, H1; Cordewener, J2;
showing consistency for the modication
America, T2; Westphal, A3; van den Hout, R1
1
HAL Allergy BV, Development, Leiden, Netherlands; of the Dp and Df extract were accom-
2
Plant Research International, Wageningen, plished.
Netherlands; 3Department of Biochemistry,
Wageningen University, Wageningen, Netherlands
Background: D. pteronyssinus (Dp) and D. 1343
farinae (Df) mite allergen extracts are trea- Stability studies on alum-adsorbed mite
ted separately with glutaraldehyde to allergoid with physicochemical tech-
obtain Dp and Df mite allergoids, respec- niques
tively. The allergoids are drug substances Luykx, D1; Cordewener, J2; America, T2; Westphal, A3;
for mite allergy vaccine PURETHAL Koppelman, S4; van den Hout, R1
mites and characterisation of these allerg-
1
HAL Allergy BV, Development, Leiden, Netherlands;
oids is needed. This includes identication
2
Plant Research International, Wageningen,
Netherlands; 3Department of Biochemistry,
of the relevant allergens in the allergoid Wageningen University, Wageningen, Netherlands;
and showing consistency of the modica- 4
HAL Allergy BV, Research and Development, Leiden,
tion process. Therefore, three successive Netherlands
Dp and Df mite extracts and corresponding
Background: Several physicochemical tech-
allergoids were analysed with various phys-
niques have been selected to investigate
icochemical techniques; mass spectrometry
their suitability to monitor the stability of
(MS), SDS-PAGE, HPLC-SEC, uores-
PURETHAL mites (PUR-M). These
cence and free lysine determination.
techniques are mass spectrometry (MS),
Method: MS: Preparation of tryptic
circular dichroism (CD), and front-face u-
digests, peptide separation via nano-HPLC
orescence (FF-uorescence). They should
before electrospray ionisation. Ionised pep-
be able to investigate the adsorbed proteins
tides were fragmented revealing amino acid
on all structural levels, i.e. primary struc-
sequences. SDS-PAGE: 1020% gels com-
ture (for protein identication), secondary
bined with Coomassie Blue staining and
and tertiary structure (for structural stabil-
reduced samples were used. HPLC: GF250
ity). Samples were temperature stressed to
SEC column was used combined with UV-
nd out if these techniques are stability
detection. Fluorescence: Emission spectra
indicating.
were recorded from 290500 nm, with exci-
Method: PUR-M: D. pteronyssinus and D.
tation at 280 nm. Lysine determination:
farinae mite extracts are modied with
Samples were hydrolysed into amino acids
glutaaraldehyde and adsorbed onto alu-
followed by free lysine determination with
minium hydroxide. MS: Preparation of
HPLC.
tryptic digests, peptide separation via
Result: A consistent group of peptides was
nano-HPLC before electrospray ionisation.
identied in the Dp and Df allergoids with
Ionised peptides were fragmented revealing
MS. These peptides were corresponding to
amino acid sequences. CD: Far-UV CD
relevant mite allergens group 1 and 2.
spectra were recorded from 195 to 260 nm
SDS-PAGE (qualitative) showed for all
while stirring the sample suspension. FF-
allergoid batches formation of various high
uorescence: Fluorescence emission spectra
molecular weight molecules via a smear
were recorded from 290 to 400 nm, with
and clear band around 250 kDa (i.e.,
excitation at 280 nm.
Mw 250kDa) on gel. HPLC-SEC (quan-
Result: Via MS relevant allergens (Der p1,
titative) showed for all allergoid batches
Der p2, Der f1 and Der f2) and other aller-
formation of molecules 670 kDa. For the
gens were identied in different PUR-M
extracts a minority of HPLC peaks is cor-
batches that were stored at different
responding to proteins with a molecular
(stressed) temperatures. The CD-spectrum
mass above 17 kDa (2230%). For the al-
of PUR-M represented a protein structure
lergoids the majority of peaks (8086%) is
consisting of mainly a-helices and b-struc-
corresponding to proteins with a molecular
ture. The FF-uorescence spectrum showed
mass (far) above 17 kDa. Determination of
an emission maximum at 326 nm suggest-
free lysines showed a consistent degree of
ing strong hydrophobic surroundings for
cross-linking in the allergoids; 4755% ly-
the aromatic amino acids present in the
sines were modied. Fluorescence studies
protein sample. Storage at various elevated
indicated a consistent hydrophobic tertiary
temperatures resulted in spectral alterations
protein structure after modication; a con-
meaning structural alterations of protein.
siderable shift of the uorescence emission
CD spectra indicated partially loss of a-
maximum (~20 nm) and a decrease of
helices (increase of ratio 207/222 nm) while
intrinsic uorescence intensity was
uorescence spectra indicated formation of
observed.
a more rigid structure (increase of uores-
Allergy 66, Suppl. 94 (2011): 482642  2011 John Wiley & Sons A/S
Poster Session 54 Immunological advances in allergen immunotherapy
cence at 326 nm). Spectra of different
PUR-M batches were very similar suggest-
ing structural similarity and consistency of
drug product.
Conclusion: MS, CD and FF-uorescence
were able to characterise the complex mite
vaccine on all structural levels, i.e. primary,
secondary and tertiary structure. Via MS
the mite allergens could be identied while
via CD and uorescence the protein struc-
ture could be investigated. By analysing
temperature stressed products these tech-
niques were identied as stability indicating
assays and applicable for showing batch-
to-batch consistency.
1344
Are all toll-like receptor (TLR) ligands
potential adjuvants in allergy vaccines?
Deifl, S1; Kitzmuller, C1; Zlabinger, G2; Steinberger, P2;
Bohle, B1
1 1
Christian Doppler Laboratory for Immunomodulation,
Department for Pathophysiology and Allergy Research,
Medical University of Vienna, Vienna, Austria; 2Medical
University of Vienna, Institute of Immunology, Vienna,
Austria
Background: Toll-like receptor (TLR)
ligands are promising adjuvants in vaccines
for allergen-specic immunotherapy. So
far, monophosphoryl lipid (MPL) A, a
TLR4-ligand, and oligodeoxynucleotides
containing CpG motifs (CpG-ODN), a
TLR9-ligand, have been tested in clinical
trials. We compared these agents with a-
gellin, a TLR5-ligand, Pam3CSK4, and
FSL-1, two TLR2-ligands, and lipopolysac-
charide (LPS), a TLR 4 ligand regarding
their effects on cells from allergic donors.
Method: Peripheral blood mononuclear
cells (PBMC) were stimulated with the dif-
ferent TLR-ligands in the presence or
absence of autologous plasma and the lev-
els of pro- and anti-inammatory cytokines
were assessed in supernatants. Monocyte-
derived dendritic cells (mdDC) were tested
for upregulation of maturation markers
and co-stimulatory molecules in response
to the TLR ligands and their functional
activity was tested in mixed leukocyte reac-
tions (MLR). mdDC were co-cultured with
CD4+CD45RA+ T cells in the absence
or presence of the different TLR ligands to
check for their potential to induce Th1- or
Th2-like responses. Finally, the inuence
of TLR ligands on allergen-uptake by
monocytes was analysed using uorescent-
ly-labelled Bet v 1.
Result: All TLR ligands induced moderate
levels of IFN-gamma but only LPS and
MPL-A induced high levels of IL-10 in
PBMC. All TLR-ligands except for CpG-
ODN induced TNF-alpha with LPS as
strongest stimulus. The presence of autolo-
gous plasma elevated all cytokine levels.
Allergy 66, Suppl. 94 (2011): 482642  2011 John Wiley & Sons A/S
Intracellular cytokine staining identied
monocytes as the main cytokine source in
PBMC. All TLR ligands induced the matu-
ration of functional DC as revealed by
MLR. Compared to LPS-matured mdDC
(Th1-promoting conditions) and mdDC
cultured with the major birch pollen aller-
gen Bet v 1 (Th2-promoting conditions),
FSL-1 and CpG-ODN promoted Th1-like
responses. The other TLR ligands upregu-
lated IL-5, IL-13, and IFN-gamma produc-
tion of nave T cells. All TLR ligands
except for CpG-ODN enhanced the uptake
of allergen.
Conclusion: Our study shows that different
TLR ligands exert different effects on cells
from allergic donors. Thus, not all TLR-
ligands are equally well suited as adjuvants
in allergy vaccines.
1345
Sublingual allergen immunotherapy:
mode of action and relationship with the
safety profile
Calderon, M1; Demoly, P2; Moingeon, P3; Malling, H4;
Simons, E5
1
Imperial College London, London, United Kingdom;
2
University Hospital of Montpellier, France;
3
Stallergenes SA, Antony, France; 4National University
Hospital, Copenhagen, Denmark; 5Department of
Pediatrics & Child Health, Department of Immunology,
Faculty of Medicine, University of Manitoba, Canada
Background: Sublingual allergen immuno-
therapy (SLIT) can be used at home and
has a better safety prole than subcutane-
ous allergen immunotherapy (SCIT), which
has a low but not negligible risk of severe/
fatal anaphylactic reactions.
Method: We performed a comprehensive
analysis of the scientic literature on SLIT
safety prole and mechanisms of action.
Considerable evidence that SLITs safety
prole is related to its mucosal mechanism
of action was found.
Result: Primarily local, oral-mucosal
adverse reactions to SLIT occur in 40
75% of patients but generally do not
necessitate dose adjustment or treatment
interruption. A recent, comprehensive
review found a rate of one SLIT-related
serious adverse event per 384 treatment
years. Only six cases of non-fatal anaphy-
lactic SLIT-related reactions are found in
papers published in indexed peer-reviewed
medical journals. Interpretation of these is
complicated by variations in the denition
and diagnosis of anaphylaxis. These
reports involved non-standardized extracts,
allergen mixtures, rush protocols, overdos-
ing or patients who had previously discon-
tinued SCIT due to severe reactions. In
oral mucosal immuno recognition, aller-
gens bind rapidly to epithelial cells and
cross the mucosa within 1530 min. Lin-
gual/oral antigen-presenting cells (APCs,
mostly Langerhans cells and myeloid den-
dritic cells) exhibit a tolerogenic phenotype
and do not typically engage in the pro-
inammatory immune responses which can
trigger systemic reactions. Oral subepitheli-
al tissues contain low numbers of mast
cells and eosinophils. SLIT-associated
immune responses include the induction of
circulating, allergen-specic Th1 and regu-
latory CD4+ T cells, leading to the estab-
lishment of clinical tolerance.
Conclusion: Rapid allergen capture by
local, tolerogenic APCs may explain
SLITs excellent safety prole.
1346
Long-term induction of IgG antibodies
induced by birch pollen-specific
immunotherapy
Mobs, C1; Ipsen, H2; Slotosch, C1; Wurtzen, P2; Hertl,
M1; Pfutzner, W1
Department of Dermatology and Allergology, Philipps
University Marburg, Marburg, Germany; 2ALK-Abello,
Experimental Immunology, Hrsholm, Denmark
Background: For a century allergen-specic
immunotherapy (SIT) is still the only
accepted and effective therapy to treat
immediate type allergies. The modications
induced by SIT are characterized by differ-
entiated immune reaction patterns. On the
cellular level the correction of an imbal-
ance between allergen-specic T helper
(Th) cell subsets and allergen-induced IL-
10-producing type 1 regulatory T (Tr1)
cells is considered to be the most critical
event in the early phase of SIT. Focusing
on SIT-induced long-term effects the
induction of allergen-specic IgG antibod-
ies (ab), especially of the IgG4 isotype,
seems to be crucial in promoting allergen
tolerance by SIT.
Method: To better understand the role of
humoral changes induced by SIT, we mea-
sured birch pollen-specic serum immuno-
globulins. Specic concentrations of IgE
and IgG, i.e. IgG4, ab for up to 66 months
were calculated after initiation of SIT by
the Phadia ImmunoCap system. To ana-
lyze potential functional implications we
investigated the inuence of SIT-induced
allergen-specic IgG ab on IgE binding to
allergen, histamine release (HR) from ba-
sophils and allergen uptake mediated by
the low afnity IgE-receptor CD23 on
antigen-presenting cells.
Result: Allergen-specic IgG and IgG4
serum levels showed a continuous increase
over the 3-year-period of SIT. After cessa-
tion of SIT, IgG serum concentrations
stopped to rise and started gradually to
decline, but still remained signicantly ele-
vated above pre-treatment levels, while no
substantial changes of IgE ab titers were
found. By analysis of serum samples
511

obtained from 12 SIT-treated birch pollen
allergic individuals we could demonstrate
that SIT-induced increases of IgG and
IgG4 ab were accompanied by enhanced
inhibition of allergen binding to IgE,
diminished HR and reduced allergen
uptake by B cells. After ending of SIT, the
concomitant decrease of IgG and IgG4 ab
titers were paralleled by diminished IgE-
blocking effects.
Conclusion: SIT-induced allergen-specic
IgG ab seem to play an important role in
establishing allergen tolerance, probably by
competing with IgE ab for capturing spe-
cic allergens. Long-term studies following
the post-treatment course of SIT are neces-
sary to determine for how long IgG ab
concentrations can be maintained at a level
high enough to block IgE-allergen interac-
tions, thereby ensuring continued allergen
tolerance.
1347
The effect of preseasonal grass pollen
allergoid immunotherapy on basophil
CD203c expression
Kepil Ozdemir, S1; Sin, B1; Guloglu, D2; _Ikinciogullari,
A2; Misirligil, Z1
1
Department of Allergy and Immunology, Ankara
University School of Medicine, Ankara, Turkey;
2
Department of Pediatric Immunology and Allergy,
Ankara University School of Medicine Ankara, Turkey
Background: Short-term preseasonal immu-
notherapy has shown to be effective in pol-
linosis. To the best of our knowledge,
there were no published studies that evalu-
ate the impact of the allergoid immuno-
therapy on CD203c expression of
basophils. We sought to assess the impact
of preseasonal grass pollen allergoid immu-
notherapy on basophil CD203c expression
and its clinical efcacy in patients with
grass pollen related allergic rhinitis.
Method: In this randomized, double-blind,
placebo-controlled study, 31 patients with
allergic rhinitis sensitized only to pollens
(conrmed by compatible history, and skin
prick tests) received either grass pollen al-
lergoids in a preseasonal immunotherapy
protocol (n = 16) or placebo injections
(n = 15). CD203c expressions were mea-
sured using ow cytometer after in-vitro
stimulation with four different concentra-
tions of grass pollen extract. The concen-
trations of the allergen extracts were
expressed as the content of the major aller-
gen Phl p 5. Basophil activation tests via
CD203c were carried out immediately
before and after the preseasonal immuno-
therapy/placebo periods and at the peak
pollen season. Symptom free days, medica-
tion free days and average visual analogue
scores during the pollen season were com-
pared to evaluate clinical efcacy.
512
Poster Session 54 Immunological advances in allergen immunotherapy
Result: The changes from baseline values
(CD203c) were signicantly different
between the two groups after in-vitro stim-
ulation with 4.510)4 lg/ml, 4.510)5 lg/
ml Phl p 5 (P < 0.05). The CD203c
expressing activated basophil percentages
after in-vitro stimulation with 4.510)4 lg/
ml, 4.510)5 lg/ml Phl p 5 signicantly
decreased compared to baseline in the
immunotherapy group (P < 0.025). There
were no signicant differences in the
CD203c expressing activated basophil per-
centages compared to baseline in the pla-
cebo group (P > 0.025). Symptom free
days were signicantly higher and average
visual analogue scores were signicantly
lower in the immunotherapy group
(P < 0.05). Medication free days were sim-
ilar between the two groups (P > 0.05).
Conclusion: This study indicates that pre-
seasonal short course allergoid immuno-
therapy is an effective method in reducing
symptoms of grass pollen related allergic
rhinitis. Furthermore, a suppressive effect
of the administered immunotherapy proto-
col was detected on the CD203c expression
of activated basophils. This study was
funded by The Scientic and Technological
Research Council of Turkey (project num-
ber 109S251).
1348
Predicting treatment response after one
course of short term immunotherapy ad-
juvanted with Monophosphoryl lipid A
Christmann, M; Girod, K; Rosewich, M; Rose, M;
Schulze, J; Schubert, R; Zielen, S
Department of Allergology, Pneumology and Cystic
fibrosis, J.W. Goethe-University, Childrens Hospital,
Frankfurt, Germany
Background: After several years of specic
immunotherapy (SCIT) there is an increase
in specic bronchial tolerance. Aim of the
study was to nd biomarkers predicting a
favourable treatment response already after
one course of short-term immunotherapy
with Monophosphoryl lipid A (Pollinex
Quattro).
Method: 22 children were included in the
study (19 male, average 10.8 years). In
bronchial allergen challenge (BAP) all had a
drop in FEV1 of >20% (mean 32.8%)
before treatment. All patients received a
short-term SCIT for grass pollen. 4 weeks
and one year after the rst treatment cycle
BAP was repeated. According to the results
patients were divided into treatment
responders and non-responders. Biomarkers
collected before SIT were evaluated in order
to predict a favourable treatment response.
Result: Four weeks after the rst treatment
cycle n = 11 children showed an improve-
ment in BAP with a drop in FEV1 < 20%
(drop out n = 3). One year after treatment
Allergy 66, Suppl. 94 (2011): 482642  2011 John Wiley & Sons A/S
n = 7 children still had a favourable treat-
ment response. The analysis of baseline bio-
markers showed signicant higher levels of
specic IgE for grass pollen in non-
responders (P < 0.01) and lower IgG4
antibody levels (5.05 versus 11.7 U/ml) after
one year. In addition, responders showed
an increase of blocking IgG4 antibodies of
119% (non-responders 5%). The only sig-
nicant marker for a favourable short-term
treatment response after 4 weeks was a
lower eNO at baseline (P < 0.05). No dif-
ferences could be seen in baseline age,
FEV1, total IgE and bronchial hyperre-
sponsiveness in methacholine challenge.
Conclusion: The majority of our patients
showed a signicantly positive effect on
bronchial tolerance 4 weeks after treat-
ment. This effect was weakened 1 year
after treatment and encourages the contin-
uation of SCIT for a second and third
cycle. Despite a small sample size our
results suggest the measurement of specic
IgE for grass pollen and the increase of
IgG4 antibodies after the rst therapy cycle
to identify treatment responders. Studies
with larger populations are necessary to
conrm our ndings.
1349
A European pattern of Der p 1 and
Der f 1 allergen exposure in a house dust
mite allergic population
Mosbech, H1; Lolck, R2; Ljrring, C3; Backer, V3
1
Gentofte Hospital, Allergy Unit, Hellerup, Denmark;
2
Djibouti; 3Denmark
Background: Population exposure to differ-
ent house dust mite (HDM) species varies
considerably in different regions. We inves-
tigated the HDM species exposure in a
HDM allergic population from eight coun-
tries in Europe.
Method: Data were obtained from a
randomised, double-blind, placebo-con-
trolled, Phase II/III trial assessing the ef-
cacy and safety of the HDM allergy
immunotherapy tablet (AIT), which con-
tains allergens from D. pteronyssinus and
D. farinae (ALK, Denmark). Participants
were 14 years of age with HDM allergic
asthma and rhinitis. HDM sensitisation
was documented by skin prick test (3 mm
weal area) and sIgE (class 2) against D.
pteronyssinus and D. farinae. Dust samples
were collected by vacuum cleaning of mat-
tresses in homes of participants from eight
European countries, and analysed for their
content of major allergen group 1 of D.
pteronyssinus (Der p1) and D. farinae (Der
f1) (MSD FORMAT V assay). Detection
limits were 4.6 ng/g dust for Der p1 and
19.6 ng/g dust for Der f1. Descriptive sta-
tistics were applied.
Poster Session 54 Immunological advances in allergen immunotherapy
Result: A total of 499 dust samples were
analysed; 470 (94%) had detectable levels
of Der p1 and/or Der f1. For the countries
GE, IT, PL and UK, >10% were below
detection limits. The highest total exposure
(Der p1 and Der f1) was found in ES
(median: 8.369 lg/g dust) and the lowest in
UK (5 lg/g). Both Der p1 and Der f1 were
detectable in the majority of samples for
the countries DK (77%), FR (69%), GE
(57%), IT (66%), PL (67%) and ES
(67%). Der f1 dominated in all countries
except UK: ranged from 62 lg/g Der f1
versus 0 lg/g Der p1 in SE to 4 lg/g Der
f1 versus 0.1 lg/g Der p1 in GE. Der p1
only dominated in UK (0.2 lg/g Der p1
versus 0 lg/g Der f1). Highest Der p1 lev-
els were found in ES and FR.
Conclusion: In this large European survey,
both D. pteronyssinus and D. farinae aller-
gens were at detectable levels in the majority
of dust samples from homes of HDM aller-
gic patients. In accordance with ndings in
The European Community Respiratory
Health Survey II, a tendency of higher levels
of Der p1 in warmer countries/close to the
Atlantic was seen. We found dominating
levels of Der f1 in all countries except UK.
However, both Der f1 and Der p1 were
present in signicant amounts in the major-
ity of countries. In order to estimate HDM
exposure in Europe, the importance of anal-
ysing for the content of both Der p1 and
Der f1 in dust samples is emphasized. These
ndings also support the inclusion of both
species in immunotherapy products such as
HDM AIT.
1350
Inhibition of grass allergen-specific T-cell
responses by CD4+CD25+CD127lo
regulatory T-cell
Wurtzen, P1; Aasbjerg, K2; Grauert, G1; Koed, G1;
Backer, V2; Lund, K1
1
ALK-Abello, Experimental Immunology, Hrsholm,
Denmark; 2Department of Respiratory Medicine,
Bispebjerg Hospital, Copenhagen, Denmark
Background: Regulatory T-cells play a cen-
tral role in regulating immune responses
and might be a key element in the effect of
allergen immunotherapy. Currently, ow
cytometric identication and sorting of
these cells are based on the surface mark-
ers CD25 and CD127 as well as the tran-
scription factor foxp3. We intend to
investigate changes in the regulatory T-cell
population during immunotherapy and
study the effect of CD4+CD25+CD127lo
cells on T-cell responses to Phleum pra-
tense allergen extract (Phl p).
Method: Percentages of CD4+CD25+
CD127lofoxp3+ cells in peripheral blood
were determined by ow cytometry. CD4+
T-cells from 11 grass allergic donors were
Allergy 66, Suppl. 94 (2011): 482642  2011 John Wiley & Sons A/S
isolated by magnetic beads purication,
and CD4+CD25+CD127lo (Treg) and
CD4+CD25+CD127high (Teff) cells were
subsequently separated by FACS. Phl p-
specic T-cell proliferation in Co-cultures
of Teff and Treg at different ratios was
investigated by EdU incorporation.
Result: The average percentage of
CD4+CD25+CD127locells was 2.8%
(1.0), and the percentage ranged from
1.2% to 4.8% among the grass allergic
donors investigated (n = 20). The average
percentage of CD4+CD25+CD127lo-
+
foxp3 cells was slightly lower (2.4%
1.0) while somewhat higher average per-
centages were found for CD25+foxp3+
and CD127lofoxp3+ (4.8% 1.7 and
4.4% 1.8, respectively). Expression of
CTLA-4 and GITR could not be demon-
strated for the majority of regulatory T-
cells while expression of both these mole-
cules was found on other T-cell subpopula-
tions. Strong inhibition of Phl p-specic
proliferation was observed in most donors,
while more modest or even absent inhibi-
tion was found in some donors (n = 11).
A link between percentages of CD4+
CD25+CD127lo T-cells in peripheral blood
and the level of Treg suppression of the
allergen-specic responses was not found
in this patient subset.
Conclusion: The average percentages of
regulatory T-cells in peripheral blood dif-
fers considerably depending on the FACS
markers used to dene this cell population.
Markers involved in the regulatory func-
tion, such as CTLA-4 and GITR were in
general not detected on the investigated
populations of regulatory T-cells in periph-
eral blood. The CD4+CD25+CD127lo T-
cells seem to function as effective inhibitors
of allergen-specic responses in grass aller-
gic patients. It will be highly relevant to
investigate the number, phenotype and
effect of these cells in detail in patients
treated with grass allergen immunotherapy.
1351
Responses of patients with multiple drug
resistant pulmonary tuberculosis to cellu-
lar immunotherapy with autologous
monocyte-derived dendritic cells
Titov, L1; Hancharou, A1; Skrahina, A1; Novokhatsko, T1;
Shpakovskaya, N2; Antonova, N2; Zalutskaya, A2;
DuBuske, L3
1
Republican Scientific and Practical Center for
Epidemiology and Microbiology, Minsk, Belarus;
2
Republican Scientific and Practical Center for
Pulmonology and Physiology, Minsk, Belarus;
3
Immunology Research Institute of New England,
Gardner, MA, United States
Background: Dendritic cells (DC) have an
important role in the initiation and control
of immune response to Mycobacterium
tuberculosis infection. Patients with multi-
drug resistant (MDR) tuberculosis (TB)
have impaired DC functions, including
maturation, antigen-processing and prim-
ing of T-cells. This study evaluates the
response of TB patients to treatment with
autologous DC.
Method: Monocytes were isolated from
peripheral blood of 13 patients with MDR
TB and cultured in AIM-V medium sup-
plemented with GM-CSF and IL-4 for
7 days to produce immature DC (iDC).
iDC were pulsed with M. tuberculosis ly-
sates obtained from each patient and cul-
tured with TNF-alpha and dibutyryl
cAMP for 1518 h to produce mature DC.
DC were assayed for immunophenotype
and sterility and injected subcutaneously
three times at 2- to 3-week intervals. Clini-
cal monitoring, sputum assessments, chest
X-rays and immune status were performed
before and 23 months after the treatment.
The control group (C) consisted of 13
patients with MDR TB matched by sex
and age.
Result: DC obtained from all patients were
sterile, viable, morphologically intact and
phenotypically mature (expression of CD83
and CD86 being > 90%). The number of
injected DC averaged 15.8 (range: 12.0
18.7) 106. Treatment with DC was well
tolerated by all patients. Combination of
standard treatment and DC promoted the
decrease or complete clearance Mycobacte-
ria TB from sputum (DC-treated patients
69.23 12.8% versus 30.76 12.8% in
C, P < 0.05) and X-ray improvement
(69.23 12.8% versus 30.76 12.8% in
C, P < 0.05). The patients treated with
DC (but not C) exhibited increased CD4/
CD8 ratios and activation of Th1 immune
responses after the trials. The number of
INF-c producing T-cells in response to M.
tuberculosis lysates increased signicantly
after treatment (before 1.6 (range 0.9
2.8), after 3.2 (range 2.33.6),
P = 0.006) reecting the accumulation of
antigen-specic T-cells in peripheral blood.
Conclusion: Treatment of TB patients with
autologous DC was safe and well tolerated.
A brief time to efcacy of DC-based treat-
ment was shown in 69% of patients versus
31% in C. DC-based treatment may
become an effective valuable method of
cellular immunotherapy for MDR TB
patients.
513

Novel mechanisms in pathogenesis of asthma
1352
Changes in peripheral blood monocyte
subpopulations in house dust mite aller-
gic asthma patients during allergen chal-
lenge
Kowal, K; Moniuszko, M; Dabrowska, M; Bodzenta-
Lukaszyk, A
Medical University of Bialystok, Allergology and
Internal Medicine, Bialystok, Poland
Background: Monocytes/macrophages play
an important role in the regulation of
inammatory response and tissue remodel-
ing. A monocyte subpopulation CD14++
CD16+CD163++ exert predominantly
anti-inammatory effects. The aim of this
study was to evaluate changes of individual
monocyte subpopulations in peripheral
blood of house dust mite allergic asthma
patients (HDM-AAs) during allergen chal-
lenge.
Method: Twenty HDM-AAs and 10 HDM
allergic rhinitis patients (HDM-ARs) with-
out asthma were included to this study.
Blood samples were collected before (T0),
6 h (TLAR) and 24 h (T24) after allergen
challenge. Blood samples were stained with
labeled monoclonal antibodies against
CD14, CD16 and CD163 and ow cytome-
try was performed using FACSCalibur cy-
tometer.
Result: The mean baseline number of cir-
culating monocytes did not differ between
HDM-AAs and HDM-ARs (438
133 cells/mm3 versus 380 84 cells/mm3;
P = 0.24). Neither in HDM-AAs nor in
HDM-ARs signicant change in the mean
number of circulating monocytes was dem-
onstrated at TLAR or T24. However, at
T0 the mean number of CD14hi++
CD16+CD163++ monocytes was higher
in HDM-AAs (141 99 cells/mm3) than
in HDM-ARs (81 48 cells/mm3, P =
0.049). At T24 signicant increase in the
number of CD14++CD16+CD163++
monocytes was demonstrated in HDM-
AAs but not in HDM-ARs. However,
when individual subpopulations of mono-
cytes where calculated as fraction of the
whole population, a signicant increase
in the number of CD14++CD16+
CD163++ monocytes in peripheral blood
was demonstrated at TLAR only in those
HDM-AAs who did not develop late asth-
matic response (LAR) but a decrease was
seen in those who develop LAR.
514
Poster Session 55
Conclusion: Allergen induced asthmatic
response is associated with changes in the
number of circulating CD14++
CD16+CD163++ monocytes. In HDM-
AAs the anti-inammatory monocyte sub-
population CD14++CD16+CD163++
seems to participate in the regulation of
asthmatic response to allergen challenge.
1353
Changes in bronchial hyperreactivity dur-
ing follow-up of adult asthmatics
Yun, I1; Lim, H2; Lee, J1; Park, J1; Hong, C1
1
Yonsei University College of Medicine, Division of
Allergy & Immunology, Seoul, Republic of Korea;
2
Yonsei University College of Medicine, Department of
Research Affair, Seoul, Republic of Korea
Purpose: To investigate any differences in
changing patterns of bronchial hyperreac-
tivity during follow-up among asthma
groups.
Methods: Sixty-one patients (22 males and
39 females; mean age, 46.3 years) who vis-
ited the out-patient clinic for bronchial
asthma from 2000 to 2002 were enrolled in
this study. They were followed up until
2008 to 2010. During the treatment period,
the patients underwent a methacholine
bronchoprovocation test at least three
times at 1- to 2-year intervals.
Results: We compared the changing pat-
terns of serial PC20s methacholine (PC20
Mch) among groups of adult asthmatics.
We grouped the patients by the onset age
of asthma, by aspirin intolerance and skin
reactivity of allergens, and by onset age
and skin reactivity of allergens. There were
no statistically signicant differences in
changing patterns of PC20 Mch among the
aforementioned groups. However, there
were signicant differences in changing
patterns of PC20 Mch among groups
divided according to initial bronchial
hypersensitivity to methacholine. During
follow-up, the patients could be grouped
by the changing patterns of bronchial hyp-
erreactivity. Ten (16.4%) patients were
classied into the good response group, 40
(65.6%) into the fair response group and
11 (18.0%) into the poor response group.
Signicant differences among the three
groups were shown in initial PC20 Mch,
past history of childhood asthma, aspirin
intolerance and skin reactivity to aeroaller-
Allergy 66, Suppl. 94 (2011): 482642  2011 John Wiley & Sons A/S
gens. We made a regression equation to
predict the good response group. The pre-
dictive probability for the good response
group was 0.961 according to the equation.
Conclusions: In this study, we were able to
classify adult asthmatics into three differ-
ent groups regarding changing patterns of
serial PC20 Mch. We suggest that interpre-
tation of these three groups as a predictive
model of asthma treatment response
should take into account initial and follow-
up PC20 Mch.
1354
Bronchial responses to D. pteronyssinus
in asthmatic subjects without systemic
atopy
Campo, P1; Antunez, C2; Mayorga, C2; Rondon, C1;
Garcia, R1; Ruiz, M1; Melendez, L2; Rodriguez-Bada, J2;
Blanca, M1
1
Carlos Haya Hospital, Allergy, Malaga, Spain;
2
Fundacion Imabis, Allergy Research Laboratory,
Malaga, Spain
Background: Recent studies have shown
evidence of positive nasal challenges to
allergens in subjects with rhinitis without
systemic atopy (negative skin testing and
serum specic IgE). This type of local rhi-
nitis also shows inammatory changes
and local production of IgE. We present
four cases of positive bronchial challenges
to D. pteronyssinus in subjects with asthma
and no systemic atopy
Method: Four subjects with mild-moderate
perennial non-allergic asthma (NAA) and
rhinitis, with suggestive symptoms of
allergy to dust mites, negative skin testing
(prick and intradermal) and negative spe-
cic IgE to D. pteronyssinus were evaluated.
Six subjects with allergic asthma (AA)
caused by D. pteronyssinus and two healthy
subjects were included as controls. Bron-
chial challenge was performed with serial
dilutions of D. pteronyssinus extract (4 lg/
ml of Der p1). Serial metacholine challenge
and induced sputum were performed prior
to and 24 h after challenge. Cell popula-
tions were evaluated by ow cytometry.
Result: All NAA and AA cases had imme-
diate positive response to D. pteronyssinus
with 20% fall in FEV1. All NAA/AA sub-
jects had positive methacholine challenge at
baseline, and PC20 decreased in both
groups 24 h after challenge (n.s), being
negative in controls (PC20 > 16 mg/ml).
Poster Session 55 Novel mechanisms in pathogenesis of asthma
Sputum ECP was higher in AA after chal-
lenge compared to NAA and controls.
However, in both NAA cases the percent-
age of eosinophils increased after challenge.
Sputum and serum basophils were higher in
AA and NAA compared with controls.
Conclusion: We present four cases of posi-
tive bronchial responses to D. pteronyssinus
in subjects with suggestive symptoms but
with no evidence of systemic atopy. More
studies are needed to increase the number of
cases and measure inammatory mediators.
1355
Overweight and obesity reduce exhaled
nitric oxide levels in Japanese women
with asthma
Imaoka, M; Tanahashi, T; Kishikawa, R; Shimoda, T;
Iwanaga, T
Department of internal medicine, Fukuoka National
Hospital, Fukuoka, Japan
Background: Overweight and obesity have
been shown to be risk factors for asthma;
there are also numerous studies describing
a role for gender in the relationship
between obesity and asthma. The aim of
this study was to determine whether there
are gender differences in the effects of
overweight and obesity on airway inam-
mation in steroid-nave adults with asthma.
Method: The subjects comprised 236 Japa-
nese adults [81 men and 155 women, median
(range) age 52 (2088) years] with asthma
who were untreated with glucocorticoster-
oids and during attack-free periods. We
used the levels of fractional exhaled nitric
oxide (FeNO) as a marker of airway inam-
mation. The FeNO concentration was mea-
sured using the recommended online
method. We compared the levels of FeNO
between overweight/obese [body-mass index
(BMI) (the weight in kilograms divided by
the square of the height in meters) 25) and
normal weight patients (BMI, 18.524.9),
separately for men and women.
Result: In 81 men, 29 overweight/obese
patients had higher, but not statistically
signicant, FeNO levels compared with 52
normal weight patients (56.4 61.7 versus
46.5 43.8 ppb, respectively; P = 0.22).
On the other hand, in 155 women, 38 over-
weight/obese patients had signicantly
lower FeNO levels compared with 117 nor-
mal weight patients (29.0 23.6 versus
39.4 33.9 ppb, respectively; P = 0.01).
Conclusion: Our results suggest that over-
weight and obesity reduce airway inam-
mation in Japanese women with asthma.
Other factors than airway inammation
alone may be involved in the mechanistic
basis underlying the relationship between
overweight/obesity and asthma in women
with asthma.
Allergy 66, Suppl. 94 (2011): 482642  2011 John Wiley & Sons A/S
1356
Eotaxin in exhaled breath condensate of
allergic asthma patients with exercise-
induced bronchoconstriction
Zietkowski, Z; Skiepko, R; Tomasiak-Lozowska, M;
Zietkowska, E; Bodzenta-Lukaszyk, A
Medical University of Bialystok, Poland, Department of
Allergology and Internal Medicine, Bialystok, Poland
Background: Eosinophils are the key
inammatory cells in asthma, and more and
more evidence suggests their crucial role in
postexercise bronchoconstriction (EIB).
Eotaxin as the most important chemotactic
factor for eosinophils, plays an important
role in the pathogenesis of asthma.
Method: The aim of the study was to eval-
uate the changes in eotaxin levels in
exhaled breath condensate (EBC) following
intensive exercise in allergic asthmatics.
The study was performed in a group of 27
asthmatics (17 with EIB, 13 without EIB)
and nine healthy volunteers. Changes
induced by intensive exercise in the concen-
trations of eotaxin in EBC during the 24 h
after an exercise test were assessed. The
possible correlations of these measurements
with the results of other tests commonly
associated with eosinophilic airway inam-
mation were also determined.
Result: In asthmatic patients with EIB, a
statistically signicant increase in eotaxin
concentrations in EBC collected during the
rst 24 h after an exercise test with maxi-
mal increase after 6 h was revealed. A
statistically signicant correlation between
the maximum increase in eotaxin concen-
trations in EBC after exercise, and either
an increase in serum ECP or FENO 24 h
after exercise in the groups of asthmatics
with EIB, was observed.
Conclusion: Our results conrm connec-
tions between post-exercise bronchocon-
striction and airway eosinophilic
inammation. The increase of eotaxin in
asthmatic airways, by promoting the
migration and activation of eosinophils,
may play an important role in upregulation
and sustaining of the airway inammation
observed after EIB in asthmatic patients.
1357
A correspondence between clinical status
and urinary leukotriene E4 in aspirin-
induced asthma
Bochenek, G; Kuschill-Dziurda, J; Sanak, M;
Nizankowska-Mogilnicka, E; Szczeklik, A
Jagiellonian University Medical College, Department of
Internal Medicine, Krakow, Poland
Background: A systemic overproduction of
cysteinyl leukotrienes (cysLTs) is a hall-
mark of aspirin-induced asthma (AIA),
reected by leukotriene E4 (uLTE4) excre-
tion into urine. We studied possible rela-
tionship between clinical status and urinary
levels of LTE4 in patients with this distinct
phenotype of asthma.
Method: Asthmatics (128 women, 66 men,
mean age 51.3 years, SD 12.5) with aspirin
hypersensitivity conrmed by provocation
tests were enrolled into the study. Clinical
data were collected using unied question-
naire with the software MS Access.
Asthma severity was assessed and levels of
asthma control were analyzed using NAE-
PP EPR3 guideline and Asthma Control
Test (ACT). Clinical course, FEV1, asthma
treatment, nasal symptoms, presence of
nasal polyps, history of polypectomies were
among parameters evaluated. Morning
samples of urine were collected and uLTE4
was estimated by ELISA (Cayman Chemi-
cal Co., Ann Arbor, MI, USA) in unpuri-
ed urine using correction for creatinine
content.
Result: uLTE4 showed no gender differ-
ences and its levels did not correlate with
FEV1. Patients with different degrees of
asthma severity had comparable urinary
excretion of LTE4. Similarly, uLTE4 did
not discriminate patients stratied by dif-
ferent levels of asthma control. No correla-
tion was found between uLTE4 and oral
or inhaled dose of corticosteroids. Patients
with nasal symptoms, nasal polyps and his-
tory of polypectomies demonstrated signi-
cantly higher levels of LTE4 than those
not affected by proximal airways disease
(P = 0.01, P < 0.001, P = 0.02, respec-
tively).
Conclusion: In AIA single measurement of
uLTE4 shows no correspondence to bron-
chial obstruction, asthma severity and con-
trol, as well as corticosteroid treatment.
On the contrary, uLTE4 levels are closely
associated with upper airway disease,
reaching the highest values in the advanced
stages of rhinosinusitis.
1358
Airway inflammation markers and cyto-
kine production by different asthma phe-
notypes in children
Nakonechna, A1; Antipkin, J2; Umanets, T2; Lapshyn,
V2; Zadorozhna, T2; Pustovalova, O2
1
Broadgreen Hospital, Allergy Dept., Liverpool, United
Kingdom; 2Institute of Paediatrics, Obstetrics and
Gynaecology, Kiev, Ukraine
Background: Eosinophils and neutrophils
play role in different asthma phenotypes,
but there is increased evidence that T cells
and cytokines are also involved, although
the relationships between these parameters
are not clearly investigated.
The aim of our study was to investigate
the airway inammation markers and cyto-
kine production in children with different
asthma phenotypes.
515

Method: Ninety-eight children aged 6
14 years with bronchial asthma and 37 age
matched controls group were observed.
Diagnosis was made using a clinical ques-
tionnaire, physical examination and skin
prick tests (SPTs) to ten common allergens.
Lung functions and induced sputum analy-
sis were measured in all patients. The total
and antigen specic IgE and IL-5, IL-6,
IL-8, IL-10, IFN-c, TNF-a levels were
assessed by ELISA.
Result: Among investigated children
66.3% (65 patients) had atopic asthma,
which was conrmed by positive SPT at
least to one allergen and increased levels of
total and specic IgE. These children had
mild-moderate asthma (84.6%) and
induced sputum eosinophilia, which corre-
lated with skin sensitization and level of
total and specic IgE. We also noted the
correlation of these parameters with
increased levels of IL-5, IL-10 and
decreased production of IFN-c (P < 0.05).
In contrast 33.7% of children (33
patients) had non-atopic asthma with nega-
tive SPT and normal levels of total and
specic IgE. They had moderatesevere
asthma, induced sputum neutrophilia,
which correlated with increased levels of
IL-6, IL-8, TNF-a (P < 0.05).
Conclusion: The investigation conrmed
that elevated level of total and allergen
specic IgE and sputum eosinophilia
directly correlates with enhanced produc-
tion of Th2 cytokines in children with ato-
pic asthma.
Children with non-atopic asthma pre-
sented sputum neutrophilia, which corre-
lates with increased production of Th1
cytokines.
1359
The receptor factor in the secretion of
cytokines induced by toluene diisocya-
nates
Liu, P1; Tsai, L2; Ching-Hui, H2; Chen, L3; Chiung, Y2
1
Department of Microbiology, Soochow University,
Taipei, Taiwan; 2Division of medicine, Institute of
Occupational Safety and Health, New Taipei City,
Taiwan; 3Department of Microbiology and
Immunology, National Defense Medical College, Taipei,
Taiwan
Background: We previously proved Tolu-
ene Diisocyanate (TDI) induces calcium
signaling in neuronal cells. Its induction on
the changes on [Ca2+]c of cells that are
active in airway, including lung epithelial
cell, T-cell and neuroblastoma cells were
found. Recently, We observed some impor-
tant proinammatory cytokines were
released by acute TDI stimulation coupled
with the changes of [Ca2+]c in T cell-line.
TDI promoted the IL-4 release signicantly
in both Jurkat cells and white blood cells,
516
Poster Session 55 Novel mechanisms in pathogenesis of asthma
moreover, an elevation of IL-4 in the
plasma of TDI allergic workers were found
also.
Method: We used a ligand of IL-4 recep-
tor, inhibitors of it and other receptors to
deliberate the roles of different receptors in
the phenomenon described above. The
induction of the increases in the mRNA
level of IL-4 by different treatment was
also analyzed to count the weight propor-
tion of the IL-4 receptor in the following
up interactions in cytokine network. Other
receptors such as nicotinic acetylcholine
receptor and muscarinic cholinergin recep-
tor were also examined also.
Result: Different effects of ligands of IL-4
receptor and inhibitor of it were found on
the cytokine secretion stimulated by TDI.
The induction of the increases in the
mRNA level of IL-4 by different treatment
was also found. By statistical methods, an
evaluation of receptor factors of IL-4
secretion and the following up interactions
between cytokine networks were discussed.
Conclusion: We postulate that IL-4 recep-
tor is important in the coupling with the
elevation in [Ca2+]c induced by TDI and
IL-4 secretion. Other receptors still cannot
be neglected in the immediate stimulation
of TDI on cytokine secretion.
1360
Dysfunctional breathing phenotype in
adults with asthma
Ciobanu, C1; Agache, I2; Paul, G3; Barbieru, V2; Creitaru,
D2
1
Theramed Medical Center, Allergy and Clinical
Immunology, Brasov, Romania; 2Transylvania
University, Allergy and Clinical Immunology, Brasov,
Romania; 3Theramed Medical Center, Psychology,
Brasov, Romania
Background: Abnormal breathing patterns
may cause characteristic symptoms and
impair quality of life. In a cross-sectional
survey 29% of adults treated for asthma in
primary care had symptoms suggestive of
dysfunctional breathing, more likely to be
female and younger, with no differences
for severity of asthma. No clear risk fac-
tors were demonstrated for dysfunctional
breathing in asthma, nor the impact of
asthma medication was evaluated.
Method: Our study aimed to describe the
dysfunctional breathing phenotype in
adults with asthma treated in a specialized
asthma center. Nijmegen questionnaire was
administered to adult patients aged 1765
with diagnosed asthma who were receiving
treatment. Main outcome were phenotypic
traits (female sex, asthma severity, asthma
duration > 5 years, asthma control, xed
airway obstruction, fast lung function
decline, frequent exacerbator, brittle
asthma, atopy, obesity, active smoker,
Allergy 66, Suppl. 94 (2011): 482642  2011 John Wiley & Sons A/S
moderate/severe rhinitis, psychopathology,
GERD, arterial hypertension, lack of ICS,
use of LABA or LTRA) independently
associated with a score >23 on Nijmegen
questionnaire (multiple regression analy-
sis).
Result: Ninety-one adults with asthma,
mean age 35.04 1.19 years, 47 (51.65%)
females completed the questionnaire. Six-
teen (17.58%) subjects had a Nijmegen
score >23. Independent phenotypic traits
for dysfunctional breathing were psychopa-
thology (P = 0.000002), frequent exacer-
bator (P = 0.01) and uncontrolled asthma
(P = 0.000000).
Conclusion: Dysfunctional breathing is not
infrequent in asthma patients and should
be seek in asthma patients presenting with
psychopathology, frequent asthma exacer-
bations or uncontrolled asthma. Asthma
medication (ICS, LABA or LTRA) has no
risk for dysfunctional breathing.
1361
Fungal colonisation of the pharynx in
patients with persistent bronchial
asthma
Reshetnikova, I1; Fassakhov, R1; Glushco, N2;
Khaldeeva, E2; Lisovskaya, S2
1
Kazan Scientific Research Institute of epidemiology
and microbiology, Eazan Medical Academy, Kazan,
Russian Federation; 2Kazan Scientific Research Institute
of epidemiology and microbiology, Kazan, Russian
Federation
Background: The purpose of this study
was to investigate fungal colonization the
pharyngeal mucosa in patients with persis-
tent asthma, who constantly applying the
inhaled corticosteroids (iGCS).
Methods: There were examined 30 patients
with bronchial asthma (13 men and 17
women, mean age 51.8 years) using iGCS
at high and moderate doses of at least
year. Mycological study of mucous phar-
ynx (posterior wall of the pharynx, tongue
root) for nding of Candida was per-
formed on the Sabourau agar medium.
Fungi growing was carried out at 2830 C
for 48 h. To study the adhesive there was
used the model of a nitrocellulose lm with
immobilized hemoglobin.
Results: Fungi of the Candida genus were
isolated from the mucosa in 23 patients
(76.6%): Candida albicans 22 cases (of
which in one case there was marked the
combination of Candida albicans and Geo-
trihum candidum), Candida tropicalis one
case. In 73.9% of the cases intensity of the
pharynx colonization by yeastlike fungi
was high and was103, 105 CFU/ml. It
has been stated that 16 out of 22 isolated
Candida albicans cultures (72.7%) have less
vivid ability to form tube germination and
psevdomitsely in comparison with clinical
Poster Session 55 Novel mechanisms in pathogenesis of asthma
cultures, isolated from the pharynx of
patient with the diagnose pharyngomikoze,
the adhesive activity of these cultures
shoved at the middle level and made 15
32%. 8 out of 22 patients (27.3%) have
cultures with a high ability for germination
tubes formation and adhesive activity at
the level of 3545%, which corresponded
to the level of clinical cultures with high
virulence.
Conclusion: The causes of differences in
the activity of the fungus Candida in asth-
matic patients receiving iGCS, require fur-
ther study.
1362
Proliferation and survival of airway
smooth muscle cells is regulated by epi-
thelium derived chemokines
Halwani, R1; Almuhsen, S1; Beland, M2; Hamid, Q2
1
Asthma Research Chair and Prince Naif Center for
Immunology Research, college of medicine, King Saud
University, Riyadh, Saudi Arabia; 2Meakins-Christie
Laboratories, McGill University, Montreal, Canada
Background: The increase in ASMC mass
is a major structural change described in
asthma. This increase has been attributed
to ASMC hyperplasia and hypertrophy.
Recent studies have suggested a role of
chemokines in SMC migration toward the
epithelium. The objective of the current
study is to test the hypothesis that chemo-
kines (Eotaxin, RANTES, IL-8 and MIP-
1a) can increase the rate of proliferation
and enhance the survival of ASM cells.
Method: ASMCs were exposed to different
concentrations of eotaxin, RANTES, IL-8
or MIP-1a. To test for proliferation, stimu-
lated ASMC were pulsed with 3H-thymi-
dine or ASMCs were stained with BrdU
and then analyzed with ow cytometry.
Apoptosis was measured using Annexin-V
and ow cytometry. P38 MAP Kinase
pathway inhibitor was used to block path-
way activation.
Result: In a concentration-dependent man-
ner, chemokines such as Eotaxin, RAN-
TES, IL-8 and MIP-1a increased ASMCs
3H-thymidine incorporation and DNA
synthesis. IL-8, Eotaxin, and MIP-1a
decreased the number of apoptotic ASMCs
compared to the matched controls. Inhibit-
ing MAP kinase pathway using ERK1/2
kinase inhibitor blocked chemokine
induced ASM cell proliferation.
Conclusion: We conclude that chemokines
contribute to increasing ASM mass by
enhancing ASMCs proliferation and sur-
vival via the activation of Erk1/2 pathway
activation.
Allergy 66, Suppl. 94 (2011): 482642  2011 John Wiley & Sons A/S
1363
Analysis of TNF-alpha polymorphisms in
a Romanian cohort of asthmatic patients
Berghea, E1; Popa, O1; Dutescu, M2; Meirosu, M3; Bara,
C4; Popa, L5
1
University Carol Davila, Bucharest, Romania;
2
National Hematology Institute Prof. Dr. C.T. Nicolau,
Bucharest, Romania; 3ICA Research & Development,
Bucharest, Romania; 4University Carol Davila,
Bucharest, Romania; 5Grigore Antipa National Museum
of Natural History, Bucharest, Romania
Background: Tumor necrosis factor alpha
(TNF-alpha) is a proinammatory cyto-
kine that contributes to airway pathology
in asthma. Genetic studies of TNF-alpha
polymorphisms in asthma have produced
different results that may reect differences
in the ethnic background.
Objectives: The aim of this study was to
investigate the association of two known
promoter polymorphisms of TNF-alpha
gene ()238G/A and )857C/T) with asthma
and its clinical phenotypes in Romania.
Methods: Ninety-two unrelated Romanian
patients with asthma and 147 healthy unre-
lated ethnically matched controls were
involved in this study. Among patients, 50
subjects had aspirin tolerant asthma (ATA)
and 42 subjects had aspirin intolerant
asthma (AIA). All individuals were geno-
typed by TaqMan SNP Genotyping Assays
C_2215707_10 and C_11918223_10, respec-
tively (Applied Biosystems, USA). Associa-
tion tests for each polymorphism and
haplotypes frequencies estimations were
performed with the software package
PLINK v 1.07.
Results: Patients and controls groups were
in HardyWeinberg equilibrium for both
polymorphisms. No association was found
between any of the investigated polymor-
phisms and asthma. In the analysis of the
subgroups of patients according to aspirin
tolerance, the frequency of the minor allele
A of the )238G/A polymorphism was
higher in AIA patients then in controls
(4% versus 2%), but not statistically signif-
icant (P > 0.05). Two main haplotypes
were constructed, the most commune being
857C/238G (77% in patients and 79% in
controls). There was no association of
these haplotypes with the disease or with
clinical phenotypes.
Conclusion: The present study shows no
association of TNF-alpha )238G/A and
)857C/T polymorphisms and derived hapl-
otypes with susceptibility to asthma and its
clinical phenotypes in Romanian patients.
1364
Double-stranded RNA up-regulates IL-32
in primary human bronchial epithelial
cells from asthmatic subjects
Brandelius, A1; Yudina, Y1; Bjermer, L2; Persson, C3;
Morgan, A4; Uller, L1
1
Department of Experimental Medical Science, Lund
University, Lund, Sweden; 2Skane University Hospital,
Lung medicine, Lund, Sweden; 3Skane University
Hospital, Clinical Pharmacology, Lund, Sweden; 4Skane
University Hospital, ENT, Lund, Sweden
Background: Interleukin-32 (IL-32) is a
newly discovered pro-inammatory cyto-
kine that may play an important role by
linking innate and adaptive immune
responses. IL-32 is mainly produced by T
lymphocytes and epithelial cells. The air-
way epithelium is target for viral infections
that are major causes of asthma exacerba-
tions. We hypothesized that viral stimula-
tion is able to induce production of IL-32
in asthmatic bronchial epithelial cells. We
have previously demonstrated that a major
immunoregulatory cytokine, thymic stro-
mal lymphopoietin (TSLP), is produced by
virally stimulated epithelial cells; the effect
abnormally great in cells obtained from
asthmatic donors. Our objective in this
study was to compare effects of a viral
infection mimic, double-stranded RNA
(dsRNA) on gene expression of IL-32 and
TSLP in primary bronchial epithelial cells
from asthmatic subjects.
Method: Primary human bronchial epithe-
lial cells (HBECs) obtained from bronchial
brushings from asthmatic individuals
(n = 4) were grown until conuent in 12-
well plates and stimulated with the viral
surrogate molecule dsRNA (110 lg/ml)
for 3 and 24 h to induce IL-32 and TSLP
gene expression (analyzed by RT-qPCR).
dsRNA-induced gene expression was also
examined in cells pre-treated for 1 h with
dexamethasone (1 lg/ml).
Result: No increased expression of IL-32
was observed after 3 h dsRNA-treatment.
However, 24 h dsRNA stimulation mark-
edly induced IL-32 expression in HBECs
(P < 0.01) compared to unstimulated cells.
In contrast, 3 h dsRNA stimulation
increased the allergy-mediating cytokine
TSLP (30 times over control P < 0.001),
whereas by 24 h dsRNA stimulation TSLP
gene expression levels were back to control,
unstimulated cells. Dexamethasone reduced
the dsRNA-induced expression of TSLP by
60% but IL-32 levels were unaffected.
Conclusion: These data indicate that viral
stimulation increase gene expression of IL-
32 as well as TSLP. However the time course
for this effect differs between these cytokines
and so does the sensitivity to steroid treat-
ment. IL-32 may complement TSLP and
contribute to immune regulation in asth-
matic airways subject to viral infection.
517

1365
Serum level of IL-6 and IL-8 in severe and
mild asthma
Adjami, M1; Hooshiar, B2; Rafiee, A2; Ghafari, J2; Ajami,
A2
1
Immunology Department, Faculty of Medicine, Shahid
Sadoughi University of Medical Science, Tehran,
Islamic Republic of Iran; 2Immunology Department,
Faculty of Medicine, Mazandaran University of Medical
Science, Sari, Islamic Republic of Iran
Background: Asthma is an inammatory
disease of lower respiratory tract. Pro-
inammatory and inammatory cytokines
like interleukin-6 (IL-6) and Interleukin-8
(IL-8) may play a key role in severity of
asthma. This study was undertaken
whether difference in serum cytokines (IL-
6 and IL-8) could involve in pathogenesis
of asthma status (severe versus mild).
Materials and Methods: In a cross-sec-
tional study, 60 patients have investigated,
including 30 patients with severe asthma
and 30 patients with mild asthma, with
recurrent wheeze, cough, dyspnea. Asthma
disease was diagnosed based on history,
clinical examination and spirometery
(according to American Society of Lung
Disease criteria). Participants asthma
severity (severe and mild) were dened
based on symptoms and forced expiratory
volume in 1 s (FEV1). We assessed IL-6,
IL-8, immunoglobulin E (IgE) serum level
by enzyme-linked immunosorbent assay
(ELISA) methods. Blood eosinophil count
also was examined. Data was analyzed
with t-test and statistical SPSS software.
Results: The levels of IL-6 were higher
than normal range (according to Kit nor-
mal range, <2 pg/ml) and IL-8 levels were
in normal range (according to Kit normal
range, <29 pg/ml) in both groups of
patients (severe and mild), serum levels of
IL-6 in 30 patients with severe asthma were
2.2 2.09 pg/ml and 2.62 2.98 pg/ml
in 30 mild asthmatic patients. IL-8 serum
levels were 11.7 7.2 pg/ml for patients
with severe asthma and 11.08 11.49 pg/
ml for mild asthmatic patients. There was
no statistically signicant difference
between level of IL-6 and IL-8 in severe
versus mild asthma (P > 0.05). 15% of
patients with severe asthma and 10% of
mild asthma showed eosinophilia that was
not signicant (P > 0.05). The IgE level in
both groups was higher than normal
(235.1 187.26 versus 228.8 200 IU/
ml) but the difference between two group
was not statistically signicant (P > 0.05).
Conclusions: This study demonstrated that
IL-6 and IL-8 involved in pathogenesis of
asthma while they have no signicant role
in severity of disease. Also there is no cor-
relation between Eosinophil count and IgE
levels and severity of asthma.
518
Poster Session 55 Novel mechanisms in pathogenesis of asthma
1366
The impact of total antioxidant capacity
on pulmonary functions of Korean
patients with asthma
Yoon, S1; Moon, K1; Baek, S2; Kim, S1; Lee, Y1; Lee, T1;
Kwon, H1; Bae, Y1; Kim, T1; Moon, H1; Cho, Y1
1
Department of Allergy and Clinical Immunology,
University of Ulsan College of Medicine, Asan Medical
Center, Seoul, Republic of Korea; 2Department of
Biostatistics, University of Ulsan College of Medicine,
Asan Medical Center, Seoul, Republic of Korea
Background: Oxidative stress is believed to
play a critical role in the pathogenesis of
asthma. But, the impact of antioxidant
capacity on outcomes of asthmatics has
not yet been clearly dened. This study
was performed to evaluate not only cross-
sectional but also longitudinal association
between serum levels of total antioxidant
capacity (TAC) and pulmonary functions
of the asthmatic patients in Korea.
Method: One hundred and seven adult
asthmatics participated in this study from
COhort for Reality and Evolution of Adult
Asthma in Korea (COREA). All the sub-
jects have been properly managed for
2 years with optimized anti-asthma medica-
tions according to the GINA guideline.
The initial baseline demographic data and
various clinical parameters were collected
at the moment of enrollment to this study.
Pulmonary function tests have been con-
ducted for each asthma patient at baseline,
1 year and 2 year after enrollment. Serum
levels of TAC at baseline were measured
by using Trolox-equivalent antioxidant
capacity assay. The associations between
TAC levels and pulmonary functions and
other parameters were analyzed. The
patients were divided into three groups
(low, intermediate and high TAC group)
and the results of pulmonary function tests
measured at baseline, 1 and 2 years after
enrollment were compared between each
group to evaluate the longitudinal effect of
TAC levels on pulmonary functions.
Result: The levels of TAC were closely
correlated with the values of pre-bronchod-
ilator FEV1% predicted at all three time
points. (at baseline, r = 2.28, P = 0.021;
at 1 year, r = 0.221, P = 0.025; at 2 year,
r = 0.254, P = 0.009) Blood eosinophils
were also positively associated with the lev-
els of TAC. The patients with higher levels
of TAC showed shorter disease durations.
In the longitudinal analysis, overall values
of FEV1 were improved after enrollment
into this study across the three groups. The
mean value of FEV1 in the group with
high TAC levels was still signicantly
greater than those in other groups at 1 and
2 years after enrollment Although there
was no statistical signicance, the differ-
ences in values of FEV1% between base-
line and 2-year after enrollment tended to
Allergy 66, Suppl. 94 (2011): 482642  2011 John Wiley & Sons A/S
be greater in the group with higher TAC
when compared to those in the other
groups.
Conclusion: The serum antioxidant capac-
ity could be an important factor to deter-
mine pulmonary functions and higher
levels of TAC may be a feasible biomarker
representing favorable prognosis in Korean
asthmatic patients.
1367
Role of haptoglobin and its polymor-
phism in bronchial asthma
Cortez e Castro, M1; Ferreira, J2; Pereira-Barbosa, M1;
Bicho, M2; Marinho, C2
1
Allergy Department, Hospital Santa Maria- CHLN
Lisbon- Portugal, Lisbon, Portugal; 2Genetic
Department, Lisbon Medical School-Lisbon- Portugal,
Lisbon, Portugal
Background: Haptoglobin (Hp), an alpha
2-sialoglycoprotein known to bind free
hemoglobin (Hb), has been implicated in
the modulation of Th1/Th2 response.The
Hp locus is located at 16q22 chromosome,
being in humans polymorphic for the alpha
chain, that leads to three genotype vari-
ants, Hp1-1, Hp2-1, Hp2-2.
Method: One hundred and sixteen asth-
matic patients were compared with a con-
trol group (n = 50) in order to:(i)
Evaluate different Hp genotype and allelic
frequencies between the two groups;
(ii)Correlate the Hp genotype with serum
Hp levels (intermediate phenotype/ endo-
type);(iii)Correlate Hp genotype and phe-
notype with asthma susceptibility/severity.
Hp levels assayed by nephelometry and
genotypes by PAGE. Statistical analysis
was performed with PASW 18, establishing
a signicance level of P < 0.05.
Result: Hp Allelic and genotype frequen-
cies were not signicantly different between
groups (P > 0.05). In asthma, differences
were observed in Hp levels in age-groups:
<29 years presented lower Hp levels com-
pared with 30 years (P < 0.05). Addi-
tionally, Hp 2-2 asthmatics have lower
levels of Hp when compared to Hp 2-1
and 1-1 (P < 0.05). Different genotype
distribution of Hp levels was only observed
in the group 15 years (P < 0.05). Hp 1-1
asthmatic patients presented an increased
risk of 4.7 to be uncontrolled when com-
pared to Hp2-2 patients [OR: 4.7; IC95%
(1.01221.891)]. No differences in Hp levels
between asthma and control group
(137.83 51.4 mg/dl versus 123.92
51.36 mg/dl), however Hp 1-1 and 2-2 indi-
viduals presented statistical differences
between groups, being the asthmatic
patients those with lower levels of circulat-
ing Hp (P < 0.05). In the control group,
no differences were observed in Hp levels
by genotype or age group (P > 0.05).
Poster Session 55 Novel mechanisms in pathogenesis of asthma
Conclusion: Despite not having observed a
prevalence of the Hp allele 1 in asthma,
that has been extensively associated with a
Th2 prole, the data point to differences
among groups that could be related to Hp
polymorphism, contributing for a different
polarization of the innate and adaptive
immune system.
1368
Proinflammatory cytokine gene variants
and airways obstruction in asthma
Daneshmandi, S1; Pourfathollah, A2; Zahra, P3;
Heidarnazhad, H4
1
Department of Immunology, Tarbiat Modares
University, Tehran, Islamic Republic of Iran;
2
Department of Immunology, School of Medical
Sciences, Tarbiat Modares University, Tehran, Islamic
Republic of Iran; 3Asthma and Allergy Research
Institute and Department of Clinical Immunology and
Allergy, Children Medical Center, Tehran University,
Tehran, Islamic Republic of Iran; 4TB and Lung Disease
Research Cancer, NRITLD, Masih Daneshvari Hospital,
Shahid Beheshti University, Medical Sciences, Tehran,
Islamic Republic of Iran
Background: Asthma is a multifactor
inammatory disorder and chronic bron-
chial inammation resulting in airway
obstruction in various degrees. Cytokines
and other inammatory mediators are
important factors in asthma pathophysiol-
ogy and airway remodeling. In this study
we evaluated role of cytokine polymor-
phisms and in lung functions.
Method: In 81 asthmatic patients ARMS-
PCR method was used to characterize IL-6
G-174C, TNF-a-A308G, TGF-b T+869C
and IFN-c T+874A polymorphisms and
PCR-RFLP using AvaII restriction enzyme
for IL-4 C-590T. A complete clinical his-
tory, physical examination, and pulmonary
function test (PFT) in a standard fashion
were performed for all subjects.
Result: IL-6-174C allele (P = 0.045),
TNF-a-308GG genotype (P = 0.002) and
TNF-a-308G allele (P = 0.004) were asso-
ciated with reduced and TNF-a-308GA
genotype (P = 0.002) with increased
FEF25-75 value in asthmatics. IFN-c
+874AA genotype caused decrease in
FVC factor (P = 0.045).
Conclusion: This study showed that IL-6,
TNF-a and IFN-c lower producer variants
are associated with reduced pulmonary
capacities. These genetic variants maybe by
affecting on cytokine synthesis participate
in airway wall thickening and obstruction
by both structural remodeling and inam-
mation. Further studies with consideration
of local, serum and systemic examination
of cytokine proteins and other different
inammatory agents and airway pathology
will be needed.
Allergy 66, Suppl. 94 (2011): 482642  2011 John Wiley & Sons A/S
1369
meric glycoproteins composed of non-
Determination of the transcription factors covalently associated alpha and beta su-
associated with altered transcriptional bunits. The best known integrins in cells
activity in asthma related squamous cell trafcking include alpha4 and beta2 con-
carcinoma antigen 1 promoter taining molecules. We hypothesized, that
also collagen receptors, alpha1beta1 and
Karaaslan, C1; Birben, E2; Sackesen, C2; Kalayci, O2
1
Molecular Biology Section, Department of Biology,
alpha2beta1 integrins (both found by us on
Faculty of Science, Hacettepe University, Beytepe, blood eosinophils of asthmatic subjects),
Ankara, Turkey; 2Pediatric Allergy and Asthma Unit, are important in cell activation and tissue
School of Medicine, Hacettepe University, Hacettepe,
Ankara, Turkey
transmigration.
Method: We assessed the expression of
Background: We have previously reported elected very late antigen (VLA) integrin su-
four polymorphisms in linkage disequilib- bunits: alpha1, alpha2, alpha4 and beta1
rium in the promoter region of the Squa- on blood CD4 and CD8 T lymphocytes in
25 adult atopic asthmatics (mild to moder-
mous Cell Carcinoma Antigen 1 (SCCA1)
that show increased promoter activity and ate persistent asthma in stable clinical con-
are associated with asthma. The aim of the dition) and 15 healthy controls, using ow
cytometry (Beckman Coulter). Studied
present study was to determine the tran-
scription factor(s) that are responsible for groups were similar by age and sex, but
the altered transcriptional activity. differed in eosinophil blood counts, serum
ECP and IgE levels. Expression of analy-
Method: Single nucleotide polymorphisms
located in the promoter region of SCCA1 sed subunits was assessed as Median Fluo-
gene were: )918 (A/G), )888 (A/G), )877 rescence Intensity (MFI).
(T/C) and )864 (A/G). )888 (A/G). Public Result: Expression of both alpha4 and
databases were searched to determine the beta1 on CD4 T cells was signicantly
higher in asthma subjects as compared to
transcription factor(s) that show >85%
healthy controls (P = 0.02 and 0.0004,
binding afnity to the sequences of inter-
respectively). Subunit alpha1 was practi-
est. Binding avidity of the mutant and wild
cally absent from blood lymphocytes (only
type genotypes were tested using TransAm
a few asthmatics showed low, detectable
ELISA before and after stimulation with
signal of this integrin). On the other hand,
IL-4 and IL-13 in nuclear extracts obtained
alpha2 chain almost absent on lympho-
from A549 cells.
cytes of healthy voluntaries, was detected
Result: Search for public databases identi-
in some asthma patients. Surprisingly, in
ed GATA-1, GATA-2 and GATA-3 as
subjects suffering from asthma for longer
candidate transcription factors. GATA-2
than 4 years (n = 15) statistically signi-
and GATA-3 but not GATA-1 showed
cant overexpression of alpha2, alpha4 and
higher binding afnity for the mutant
beta1 subunits on both CD4 and CD8 T
genotype both under unstimulated condi-
cells was observed as compared to con-
tions and following IL-4 and IL-13 stimu-
trols. The same was true for patients suf-
lation compared to the wild type (GATA-
fering from persistent moderate asthma
2: P = 0.003, P = 0.043, GATA-3:
(n = 7).
P = 0.003, P = 0.018).
Conclusion: Expression of VLA integrins
Conclusion: Differential binding of GATA-
on blood lymphocytes in asthma depends,
2 and GATA-3 to the mutant and wild
among others, on asthma severity and
type sequences are responsible for the
duration. In more severe and long-lasting
altered transcriptional activity of the
disease alpha2, alpha4 and beta1 subunits
SCCA polymorphisms. ?Supported by
are overexpressed on CD4 and CD8 lym-
Turkish Scientic and Technological
phocytes. The biological role of alpha2-
Research Institute
beta1 integrin in asthma is unknown. This
integrin has been described as a stimulator
of collagen accumulation in the airways
1370 and may play a role in asthma airway
Overexpression of very late antigen inte- remodeling. It is tempting to speculate that
grins on blood lymphocytes in asthma integrin collagen receptors may become a
patients
new therapeutic target in asthmatic airway
Zuk, J; Bazan-Socha, S; Musial, J remodeling.
Jagiellonian University Medical College, Allergy and
Clinical Immunology, Krakow, Poland
Background: In bronchial asthma T lym-
phocytes possibly direct cells to the sites of
inammation. Recruitment of engaged cells
from blood to the airways is mediated by
adhesive molecules e.g. selectins and inte-
grins. Integrins are a family of heterodi-
519

1371
Circulating T-bet+, GATA-3+, RORct+ and
FOXP3+ cells in individuals with eosino-
philic and non-eosinophilic asthma
Malmhall, C; Bossios, A; Radinger, M; Sjostrand, M; Lu,
Y; Lundback, B; Lotvall, J
University of Gothenburg, Sahlgrenska Academy/
Krefting Research Centre, Goteborg, Sweden
Background: Cluster analysis has revealed
distinct groups of asthma phenotypes,
some expressing eosinophilic inammation
and some not. It is likely that different
subsets of T helper cells can be involved in
the immune process driving the eosino-
philia related asthma compared to the non-
eosinophilic asthma. Our aim was to deter-
mine whether individuals with asthma and
eosinophilia express signs of different regu-
latory immune mechanisms compared to
asthmatics without signs of eosinophilia,
and non-asthmatic controls, specically
determining the relationship of eosinophilia
520
Poster Session 55 Novel mechanisms in pathogenesis of asthma
to circulating Th1, Th2, Th17 and Treg
cells.
Method: Participants were selected from
an epidemiological cohort, the West Swe-
den Asthma Study. Immunophenotyping
of fresh peripheral blood cells obtained
from ve healthy non-asthmatics and two
groups of asthmatics, seven subjects with
and six subjects without eosinophilia
related asthma, was performed using ow
cytometry.
Result: This study shows that individuals
with asthma and concurrent eosinophilia
have signicantly increased numbers of cir-
culating Th2 cells (expressing the transcrip-
tion factor GATA-3) compared to both
controls and asthmatics without eosino-
philic inammation (P < 0.05), whereas
no differences in the number of Th1 (T-
bet+), Th17 (RORct+) or Treg
(FOXP3+) cells were observed in compari-
son to control subjects. GATA-3+ cells
Allergy 66, Suppl. 94 (2011): 482642  2011 John Wiley & Sons A/S
expressing additional transcription factors,
including T-bet and/or RORct or FOXP3,
were numerically more abundant in indi-
viduals with eosinophilia related asthma.
In comparison to control subjects, asth-
matics without eosinophilic inammation
show lower numbers of T-bet+, RORct+
and FOXP3+ cells as well as signicantly
reduced numbers of T-bet+ and RORct+
cells expressing additional transcription
factors (P < 0.05).
Conclusion: Individuals with eosinophilia
related asthma express signs of a specic
immune process involving Th2 cells in con-
trast to individuals with non-eosinophilic
asthma. The ability of human T helper cells
to express several regulatory transcription
factors argues for plasticity of these cells.
The proportion of possible less exible
T helper cells in individuals with non-eosin-
ophilic asthma may be a sign of disease.

Novel approaches to allergy diagnosis
1372
Polysensitisation to pollens analyzed and
reinterpreted with the microarray
technique
Chambel, M1; Paiva, M1; Prates, S1; Loureiro, V2; Leiria
Pinto, P1
1
Hospital Dona Estefania, Immunoallergology
Department, Lisbon, Portugal; 2Hospital Dona Estefania,
Pathology Department, Lisbon, Portugal
Background: ImmunoCAP ISAC (Immuno
Solid-phase Allergen Chip) is a new semi-
quantitative in vitro test that aims the detec-
tion and identication of seric specic IgE
(sIgE) to multiple allergenic molecular com-
ponents. Objectives: To analyze the results
of ImmunoCAP ISAC (ISAC) in patients
with respiratory allergic disease, with skin
prick tests (SPT) suggestive of polysensitiza-
tion to pollens.
Method: We selected 34 patients with
respiratory allergy and SPT positive to two
or more pollens extracts. In all patients,
sIgE to mollecular allergens were deter-
mined by the ISACmethod; val-
ues 0.3 ISU were assumed as positive.
We only analyzed the results to the pollens
tested with both methods (grasses, parie-
taria, mugwort, salsola, olive tree, plane
tree, birch).
Result: Median age was 18 years (450),
with predominance of male sex (56%). The
median number of positive SPT per patient
was 4 (29). Grasses were responsible for
the highest number of sensitizations
(n = 33) and birch for the lowest (n = 10).
With ISAC, we found that the highest
number of positivity was for grass allergens
(n = 31 patients), with birch in second
place (n = 20) and the last being plane tree
(n = 6). To all pollens, except grasses and
birch, there were a higher number of posi-
tive results with SPT than with ISAC. The
concordance between the two methods is
high to grasses (91%), ranging from 79%
to 47% to other pollens. With exception of
birch, the discordant cases mainly result
from positive SPT and negative results with
ISAC. For birch, the discordant cases
result from the opposite phenomenon.
When we analyzed the frequency of sensiti-
zation to panallergens (rPhl p7, rPhl p12,
Bet v2, Bet v4) we found that it was higher
in cases of discordance between the two
methods (compared with the concordant
cases C 79% versus 48%).
Allergy 66, Suppl. 94 (2011): 482642  2011 John Wiley & Sons A/S
Poster Session 56
Conclusion: In this sample, grasses were
the most frequent sensitizing pollen, with a
good concordance between the two meth-
ods. For other pollens, there was a consid-
erable number of patients in which ISAC
was negative although SPT were positive
to the same pollen. In a high percentage,
this phenomenon was associated with sen-
sitization to panallergens, suggesting that
these may be the responsible for the falsely
positive results in SPT, resulting in a pat-
tern of apparent polysensitization. ISAC
gives a useful contribution to discriminate
between polysensitization and cross-reac-
tion in some patients, allowing us to refor-
mulate therapeutic strategy.
1373
IgE cross-reactivity between Bet v 1 and
cytokinin-specific binding protein from
mung bean, a homologous protein out-
side the PR-10 subfamily
Guhsl, E1; Gepp, B1; Balasz, N1; Hofstetter, G1;
Hemmer, W2; Breiteneder, H1; Radauer, C1
1
Department of Pathophysiology and Allergy Research,
Medical University of Vienna, Vienna, Austria;
2
Floridsdorfer Allergiezentrum, Vienna, Austria
Background: Cytokinin-specic binding
protein (CSBP) from mung bean (Vigna
radiata) is a member of the Bet v 1 family
with low sequence identity (31%) to the
major birch pollen allergen, Bet v 1, never-
theless adopting an identical fold. The aim
of this study was to compare CSBP with
Bet v 1 regarding biochemical and immu-
nological aspects.
Methods: Percent surface identity was cal-
culated based on a structural alignment of
the crystal structures of Bet v 1 and CSBP
using an algorithm that took into account
backbone conformations and identities of
aligned residues. CSBP was expressed in
Escherichia coli and puried by metal che-
late afnity and ion exchange chromatog-
raphy. Secondary structures of rCSBP and
rBet v 1.0101 were compared using circular
dichroism spectroscopy. Binding and cross-
reactivity of IgE from Bet v 1-sensitized
patients sera to rCSBP and rBet v 1.0101
were examined by ELISA and ELISA inhi-
bition.
Results: Structural comparison of Bet v 1
and CSBP revealed that 29% of the sol-
vent-accessible surface area of CSBP was
identical to Bet v 1. In addition, two con-
served surface patches were identied as
potential cross-reactive epitopes. The CD
spectra of Bet v 1.0101 and CSBP were
nearly identical. Ten of 35 (29%) birch
pollen allergic patients sera showed IgE
binding to CSBP. IgE binding to CSBP
was completely inhibited by preincubation
of the sera with Bet v 1.0101. In contrast,
CSBP inhibited IgE binding to Bet v
1.0101 only by 1520%.
Conclusion: This study, supported by
grants P-22559-B11 (CR) and SFB-F01802
(HB) from the Austrian Science Fund,
showed IgE cross-reactivity between Bet v
1 and a homologous protein outside the
PR-10 subfamily. In addition to Vig r 1,
the PR-10 subfamily member, IgE binding
to CSBP may contribute to allergic reac-
tions to mung bean sprouts.
1374
Molecular characterisation of pollen
allergens from the pectate lyase family
Sancho, A1; Asam, C1; Hauser, M1; Bohle, B2;
Ferreira, F1
1
Department of Molecular Biology, University of
Salzburg, Salzburg, Austria; 2Department of
Pathophysiology, Medical University of Vienna, Vienna,
Austria
Background: Certain allergens found in
cypress, cedar, mugwort, and ragweed pol-
len belong to the protein family of pectate
lyases and are implicated in rhinoconjunc-
tivitis and asthma. Plant derived members
of this family are difcult to produce in a
heterologous recombinant form. However,
well dened proteins are needed for diag-
nosis and as new vaccine candidates for
allergen-specic immunotherapy. The aim
of this study is to develop the heterologous
expression of pollen pectate lyases and to
characterise the allergenic effect of natural
and recombinant forms of these proteins.
Method: Allergens were cloned, expressed
in a suitable Escherichia coli host strain
and puried by standard chromatographic
techniques. The natural counterparts were
also puried and their identity and purity
veried by N-terminal sequencing and pep-
tide mass ngerprinting. The aggregation
behaviour in solution and the secondary
structure of the puried proteins were
determined by SEC and circular dichroism
(CD)-spectroscopy, respectively. Human
521

allergen-specic IgE binding properties to
the natural and recombinant allergens were
assessed by immunoblotting and ELISA
experiments.
Result: A panel of allergens of the pectate
lyase family was either puried from the
natural sources or as recombinant forms.
CD analysis of the puried allergens indi-
cated a mixture of alpha-helical and beta-
sheet structures, which is in accordance
with the known structure of pectate lyase
proteins. The immunological properties of
the recombinant allergens were comparable
to those of the natural proteins.
Conclusion: Pollen pectate lyases can be
successfully expressed in a heterologous
host and will allow further examination of
these proteins as candidates for improved
diagnosis and for specic immunotherapy.
1375
Polcalcin IgE and clinical reactivity in
patients from the mediterranean area
Scala, E; Ferrara, R; Zennaro, D; Alessandri, C; Palazzo,
P; Pomponi, D; Liso, M; Mari, A
IDI-IRCCS, Center for Molecular Allergology, Rome,
Italy
Background: Polcalcins are a group of
highly cross-reactive allergens belonging to
the family of calcium-binding proteins
sharing common domains termed EF-
hands that are specically expressed in pol-
len tissues. Polcalcins are responsible for
IgE co-recognition between unrelated pol-
len species and considered the marker to
identify multiple pollen-sensitized patients.
The purpose of this study was to analyse
the prevalence of polcalcins IgE reactivity
within an Italian allergic population.
Method: Era from patients complaining
about allergic symptoms was studied.
Demographic and clinical information,
including symptoms and all diagnostic test
results, were recorded by means of the In-
terAll software version 3.0 (Allergy Data
Laboratories s.c., Italy). The ISAC103
allergen microarray (PMD, Austria) has
been used to test sera for allergenic mole-
cule specic IgE, including two polcalcins
from timothy grass (Phl p 7) and birch pol-
len (Bet v 4). Prevalence of respiratory
symptoms has been calculated in patients
tested positive for the polcalcins. Patients
prole for all the remaining allergens spot-
ted on the ISAC103 microarray was also
evaluated.
Result: We studied 414 patients (210 f,
mean age 33.6 + 17.3) reactive to Bet v 4
or Phl p 7. 45.9% of patients were older
than 45 years. 331 of them (79.9%) was
Bet v 4+/Phl p 7+, whereas 45 were Bet v
4-/Phl p 7+, and 38 had Bet v 4+/Phl p
7- immune phenotype. A signicant direct
relationship of IgE co-recognition of pol-
522
Poster Session 56 Novel approaches to allergy diagnosis
calcins has been recorded (P < 0.001).
Considering genuine allergens from pollen
sources, 96.8% of patients were sensitized
to timothy grass pollen allergens, 59.6% to
cypress pollen major allergen Cup a 1,
48.3% to the pellitory allergen Par j 2, and
30.7% to the platan pollens studied. Con-
current IgE reactivity to prolin or PR-10
panallergens was observed in 29.5% and
23.9% of patients, respectively. 75.8% of
polcalcin sensitized individuals complained
about rhino-conjunctivitis and 50.5% had
asthmatic symptoms.
Conclusion: Polcalcin represent an impor-
tant marker of multiple pollen-sensitized
patients. We found that polcalcins reactiv-
ity is associated with IgE recognition of
genuine molecules belonging to unrelated
pollen sources and less commonly to other
plant derived panallergens. The asthma
prevalence within the study group suggests
polcalcins as markers of severity as previ-
ously preliminary reported though clinical
data should be further conrmed.
1376
Development of an immunoassay to
quantify purified allergens based on the
competition of the binding to solid-phase
based specific IgE
Galan, A; Gavilan, D; Gomez, N; Ledesma, A; Jimeno,
L; Barber, D
ALK-Abello, Research, Madrid, Spain
Background: The in vitro standard methods
to measure allergens in extracts are mainly
based in the use of specic monoclonal
antibodies. The availability of these
reagents could be problematic in some
cases and furthermore, the specicity of
these reagents could be a problem if we
want to measure homologue allergens in
different sources. Since the determination
of specic IgE levels against puried aller-
gens using the platform ADVIA-Centaur is
a validated method with a high sensitivity
and precision, the purpose of this work has
been to use this platform to quantify puri-
ed allergens, using the panallergen prolin
as example.
Method: The assay employs a reverse
sandwich architecture using a monoclonal
anti-human IgE covalently bound to para-
magnetic particles in the solid phase cap-
turing the specic IgE from the serum
pool. Afterwards, it is incubated with the
allergen (biotinylated allergen plus sample
or standard). The amount of biotinylated
allergen captured by the IgE will be inver-
sely dependent on the amount of non-
labelled allergen present in the sample or
standard. By using a standard curve of
known concentration, the test samples can
be quantied by interpolation.
Allergy 66, Suppl. 94 (2011): 482642  2011 John Wiley & Sons A/S
Result: The assay has been applied to
quantify the panallergen prolin. A com-
petitive assay using puried Pho d 2 (palm
pollen prolin) as standard, has been
developed and the amount of prolin
equivalent has been quantied in seven
pollen extract preparations (Betula, Che-
nopodium, Lolium, Olea, Phleum, Salsola,
Plantago). The results show very high vari-
ability in the proling equivalent content,
with a range between 1 lg/vial in the Betu-
la extract and 75 lg/vial in the Lolium
preparation and also it is observed the
amount of this allergen present in these
extracts is very low compared to the con-
tent of major allergen.
Conclusion: A suitable new method to
quantify puried allergens has been devel-
oped. A signicant advantage is that the
method does not need the availability of
specic monoclonal antibodies, and since
specic IgE is used, the allergenic activity
is quantied too. As an example, this assay
has been applied for the quantication of
the panallergen proling in several pollen
extracts.
1377
The receptor factor in the secretion
of cytokines induced by toluene
diisocyanates
Chiung, Y1; Liu, P2; Tsai, L1; Chen, L3; Ching-Hui, H1;
Kao, Y2
1
Division of Medicine, Institute of Occupational Safety
and Health, New Taipei City, Taiwan; 2Department of
Microbiology, Soochow University, Taipei, Taiwan;
3
Department of Microbiology and Immunology,
National Defense Medical College, Taipei, Taiwan
Background: We previously proved Tolu-
ene Diisocyanate (TDI) induces calcium sig-
naling in neuronal cells. Its induction of the
changes on [Ca2+]c of cells that are active
in airway, including lung epithelial cell, T-
cell and neuroblastoma cells were found.
Recently, We observed some important
proinammatory cytokines were released by
acute TDI stimulation coupled with the
changes of [Ca2+]c in T cell-line. TDI pro-
moted the IL-4 release signicantly in both
Jurkat cells and white blood cells, more-
over, an elevation of IL-4 in the plasma of
TDI allergic workers were found also.
Method: We used a ligand of IL-4 recep-
tor, inhibitors of it and other receptors to
deliberate the roles of different receptors in
the phenomenon described above. The
induction of the increases in the mRNA
level of IL-4 by different treatment was
also analyzed to count the weight propor-
tion of the IL-4 receptor in the following
up interactions in cytokine network. Other
receptors such as nicotinic acetylcholine
receptor and muscarinic cholinergin recep-
tor were also examined also.
Poster Session 56 Novel approaches to allergy diagnosis
Result: Different effects of ligands of IL-4
receptor and inhibitor of it were found on
the cytokine secretion stimulated by TDI.
The induction of the increases in the
mRNA level of IL-4 by different treatment
was also found. By statistical methods, an
evaluation of receptor factors of IL-4
secretion and the following up interactions
between cytokine networks were discussed.
Conclusion: We postulate that IL-4 recep-
tor is important in the coupling with the
elevation in [Ca2+]c and IL-4 secretion
induced by acute treatment of TDI. Similar
effects on other receptors still cannot be
ignored.
Keywords: toluene diisocyanate (TDI);
cytokine; Jurkat cell; IL-4 receptor.
1378
Purification and cross-reactivity profiling
of the five clinically most relevant aller-
genic pectate lyases
Hauser, M1; Wolf, M1; Wallner, M1; Himly, M1; Ebner,
C2; Briza, P3; Mari, A4; Behrendt, H5; Ferreira, F1
1
Christian Doppler Laboratory for Allergy Diagnosis and
Therapy, Department of Molecular Biology, University
of Salzburg, Salzburg, Austria; 2Allergieambulatorium
am Reumannplatz, Vienna, Austria; 3Department of
Molecular Biology, University of Salzburg, Salzburg,
Austria; 4Center for Molecular Allergology, IDI-IRCCS,
Rome, Italy; 5ZAUM, Center for Allergy and
Environment, Munich, Germany
Background: The botanically unrelated
families of Asteraceae and Cupressaceae
are widely distributed over the Northern
hemisphere and their pollen represents one
of the most clinically relevant allergenic
sources. However, the major disease elicit-
ing allergens all belong to the same protein
family of pectate lyases. To date, recombi-
nant allergenic pectate lyases showing
native folds are not available, thus most
studies elucidating pectate lyase allergies
were performed using allergen extracts.
Method: During the present study, ve
natural allergenic pectate lyases from As-
teraceae as well as Cupressaceae pollen
(Amb a 1, Art v 6, Cry j 1, Cup a 1, and
Jun a 1, respectively) were puried to
homogeneity by standard chromatography
techniques. The optimized methods
allowed purication of structurally intact
pectate lyases without detectable auto-pro-
teolysis, degradation, or truncation prod-
ucts. Puried proteins were characterized
physicochemically in terms of identity,
structural integrity, and proteolytic stabil-
ity. Additionally, an assay to analyze the
pectolytic activity was established and the
puried pectate lysases as well as pollen
extracts were characterized in detail.
Result: Different pectolytic activities were
detected for all ve candidate allergens.
Furthermore, the puried pectate lyases
were used for a large-scale cross-reactivity
Allergy 66, Suppl. 94 (2011): 482642  2011 John Wiley & Sons A/S
study including patients sera from four
distinct geographical areas (Northern
America, the Mediterranean Area, Central
Europe, and Asia, respectively). Little
cross-reactivity has been observed between
Asteraceae and Cupressaceae pectate lyas-
es, whereas within the two families the
level of cross-reactivity was relatively high.
Conclusion: These results allow the classi-
cation of allergenic pectate lyases, which
will provide the basis for the selection of
suitable candidate molecules for diagnosis
and therapy of pectate lyase-triggered aller-
gic reactions.
1379
Strategy to obtain mature Der p 1 by
inserting a proteolytic cleavage site into
the proDer p 1 gene
Petersen, A1; Bottger, M2; Suck, R2; Jappe, U2
1
Research Center Borstel, Clinical and Molecular
Allergology, Borstel, Germany; 2Germany
Background: House dust mite allergen Der
p 1 is one of the most important aeroaller-
gens. For component resolved diagnosis
and functional research, pure and well-
dened recombinant Der p 1 is requested.
To prevent early proteolysis and self degra-
dation, the allergen must be expressed as
proDer p 1 (proenzyme). Subsequent split-
ting of the prosequence under acidic condi-
tions is critical. If cleavage occurs, the
active Der p 1 might cause self degrada-
tion.
Objective: Cloning Der p 1 with a TEV
(Tobacco Etch Virus) protease cleavage
site between the prosequence and the
mature Der p 1 sequence to express and
isolate proteolytic active Der p 1 under
controlled conditions.
Method: The TEV cleavage site was cloned
into the gene coding for proDer p 1. After
ligation into pET23b vector and transfec-
tion into E. coli BL21 cells, proDer p 1
was expressed and isolated by Ni-NTA
chromatography. Proteolytical cleavage
was tested under various conditions (pH
and reducing agents). Since prosequence
and TEV protease were His-tagged, they
could be removed by a further separation
on Ni-NTA sepharose. Eluted samples
were investigated by Western blotting using
patients sera and monoclonal antibodies
to identify Der p 1.
Result: Removal of the prosequence from
proDer p 1 under acidic conditions did not
work in our hands. Expression of the
proDer p 1 construct with a TEV cleavage
site (36 kDa) provided suitable amounts
(about 10 mg puried proDer p 1 per l).
Incubation of 100 lg proDer p 1 with
250 U TEV protease for 1 h yielded about
80% cleavage of prosequence and mature
sequence. This was estimated from Coo-
massie stained SDS gels showing protein
bands of 28 and 10 kDa, respectively, and
correlates with the theoretical molecular
masses.
Conclusion: Instead of the spontaneous
cleavage of the prosequence from the
mature Der p 1 sequence, we obtained a
precise proteolytic cleavage by integrating
a TEV cleavage site between pro- and
mature sequence. As shown by the use of
monoclonal antibodies we were able to
detect mature Der p 1. Functional assays
with the isolated mature Der p 1 will dem-
onstrate whether the product shows prote-
olytic activity.
1380
An improved method for isolation and
purification of native bovine b-lactoglobu-
lin; separation of A and B b-lactoglobulin
isoforms
Stojadinovic, M1; Burazer, L2; Ercili-Cura, D3; Sancho,
A4; Buchert, J3; Mills, C4; Cirkovic Velickovic, T1; Stanic-
Vucinic, D1
1
Faculty of Chemistry, University of Belgrade,
Biochemistry, Belgrade, Serbia; 2Institute of Virology,
Vaccines and Sera Torlak, Belgrade, Serbia; 3VTT
Technical Research Centre of Finland, Espoo, Finland;
4
Institute of Food Research, Norwich, United Kingdom
Background: b-lactoglobulin (BLG) is the
major whey protein of cows milk exten-
sively used as a functional ingredient in
food industry due to its functional and
nutritional properties, but at the same time
BLG is known as one of the major milk
allergens. Until now many methods for iso-
lation of this protein have been described.
The scope of this paper was to develop an
easy, inexpensive, one-step method for iso-
lation and purication of BLG while pre-
serving its native structure.
Method: BLG was puried from defatted
whey obtained from raw cows milk by
anion-exchange chromatography. Protein
purity and identity were determined using
reversed-phase HPLC and mass spectrome-
try. The structure and native conformation
of the isolated BLG and its potential to
bind anti-BLG rabbit antibodies were com-
pared to the standard commercial BLG by
circular dichroism (CD) spectrometry and
by inhibition ELISA, respectively. Their
susceptibility to various cross-linking
enzymes was also studied.
Result: Total BLG yield was 80% with
purity from 9799%. Far-UV CD spectra
indicated that the isolated and the stan-
dard BLG molecules had similar secondary
structures, while near-UV CD spectra
showed that the standard BLG had slightly
disrupted tertiary structure compared to
the isolated BLG. The isolated protein
bound anti-BLG rabbit antibodies with
higher afnity (IC50 = 0.163 lg/ml) then
523

the standard BLG (IC50 = 0.414 lg/ml).
Their susceptibility to cross-linking
enzymes pointed out that the puried BLG
is more compact than the standard BLG.
Conclusion: A one-step method for isola-
tion and purication of bovine BLG pre-
sented here has several advantages over
other methods: it is a cheap, efcient and
reproducible method yielding milligrams/
grams of highly puried native BLG with
the separation of A and B BLG isoforms.
Therefore isolated protein is suitable for
studying antigenic and molecular charac-
teristics of this protein, as well as the effect
of food processing on these properties.
This method is suitable for laboratory or
medium-scale isolation yet holding scale-up
potential for industrial purposes.
1381
Comprehensive allergen quantitation
using mass spectrometry
Nony, E1; Bouley, J1; Le Mignon, M1; Fenaille, F2;
Becher, F2; Chabre, H1; Lautrette, A1; Batard, T1;
Moingeon, P1
1
Stallergenes, Scientific Department, Antony, France;
2 Center of RAS, Laboratory of human molecular
Commissariat a lEnergie Atomique, Service de
Pharmacologie, Gif-sur-Yvette, France
Background: The exact quantitation of rel-
evant allergens is essential to support the
development of high-quality biologicals
intended for safe and efcacious immuno-
therapy. In this regard, conventional
ELISA assays often fail to detect selected
allergen isoforms. Herein, we describe the
application of mass spectrometry (MS) in
order to quantify major allergens present
within two pharmaceutical-grade allergen
products: a 5-grass pollen extract (from
Pooideae) and a house dust mite extract
(made from Dermatophagoides pteronyssi-
nus and Dermatophagoides farinae species).
Method: An in-depth MS characterisation
of group-1 and group-2 allergens puried
from grass pollens and house dust mites,
respectively, was rst performed to estab-
lish primary structure variability. Based
upon those results, proteotypic allergen
peptides were selected and analysed using
liquid chromatography (LC)-ESI-MS/MS.
To this aim, target tryptic peptides were
monitored in comparison with synthetic
peptides labelled with [13C6,15N]Leu and
[13C6,15N]Phe used for calibration.
Result: The primary structures of allergens
considered in the present study were
entirely conrmed, leading to the identi-
cation of numerous (i.e. up to 20) promi-
nent sequence variants. Two MS
quantitation assays were consequently
developed to monitor group-1 content in a
ve grass mix vaccine and group-2 within
a house dust mites extract, based on the
determination of three specic proteotypic
524
Poster Session 56 Novel approaches to allergy diagnosis
(i.e. highly conserved) tryptic peptides for
each assay. Quantitation results, based
upon the detection of such conserved pep-
tides, conrmed batch-to-batch consistency
and therefore process robustness. When
compared to established ELISA sandwich
methods, MS provided up to 50-fold
higher quantitation values for both group-
1 grass allergens and group-2 mite aller-
gens, conrming that ELISA method fails
to detect some isoforms.
Conclusion: Mass spectrometry represents
an effective tool for accurate and compre-
hensive quantitation of pharmaceutical-
grade allergens and may favourably substi-
tute for conventional ELISA methods.
1382
Quantitative analysis of cytokine expres-
sion in peripheral blood lymphocytes and
nasal mucosa of the patients with aller-
gic diseases
Fedorova, Y1; Karunas, A1; Nurgalieva, A2; Gimalova,
G1; Murzina, R2; Levasheva, S2; Gatiyatullin, R2; Etkina,
E2; Khusnutdinova, E1
1
Institute of Biochemistry and Genetics, Ufa Science
genetics, Ufa, Russian Federation; 2Bashkir State
Medical University, Ufa, Russian Federation
Asthma and allergic rhinitis (AR) are
complex genetic disorders that result from
interactions between multiple genes and
environmental factors. Immune and inam-
matory responses mediated by cytokines,
play important roles in the pathophysiology
of allergic diseases. These responses are
associated with overexpression of Th2 cyto-
kines. Using RT-PCR, we performed a
cytokine genes expression analysis in
peripheral blood lymphocytes and epithelial
cells from human nasal mucosa of nine
patients with asthma, seven subjects with
allergic rhinitis, and seven healthy control
subjects. A highly sensitive quantitative
real-time PCR method for IL4, IL18 and
IL13 mRNA quantication was performed
using the SYBR Green. In accordance with
delta-delta Ct method, the mRNA
amounts of a target gene were normalized
to an endogenous control (GAPDH). Nor-
mality of quantitative data was assessed
using the KolmogorovSmirnov test. T
tests were used when normality assumptions
were satised, otherwise the MannWhitney
U test was utilised. The statistical analysis
was done using the GraphPad Software and
statistical signicance was obtained with
P < 0.05. Specimens from peripheral blood
lymphocytes of patients with asthma
showed higher level of IL18 mRNA expres-
sion (M SEM: 3.22 0.76) than those
from healthy patients (M SEM:
1.34 0.41) P = 0.04. Increased level
expression of IL18 gene was detected in
Allergy 66, Suppl. 94 (2011): 482642  2011 John Wiley & Sons A/S
nasal mucosa of the patients with allergic
rhinitis (M SEM: 241.7 90.67, P =
0.005). Expression analysis demonstrated a
signicant increase in the expression of
IL13 in peripheral blood lymphocytes of
asthma patients (M SEM: 7.00 1.855)
as compared with controls (M SEM:
2.04 0.91) P = 0.04. Also IL13 was
highly expressed in epithelial cells from
human nasal mucosa of AR patients
(M SEM: 260.1 119.4, P = 0.0005).
This study provides additional evidence
that the expression of Th2 cytokine (IL18,
IL13) genes play key roles in the allergic
inammation.
1383
Structural and enzymatic characterisation
of pollen allergen Phl p 4
Zafred, D1; Nandy, A2; Keller, W1
1
University of Graz, Institute of Molecular Biosciences,
Graz, Austria; 2Allergopharma, J. Ganzer KG, Reinbek,
Germany
Background: As natural extracts carry the
risk of uncontrolled side effects, recombi-
nant proteins are becoming important
component of vaccines. Side effects of the
allergen administration can be further
reduced by introduction of hypoallergenic
derivatives. These are developed in such a
manner that they evoke the immune
response but do not cause mast cell
degranulation. In order to create such hyp-
oallergens in a rational manner the knowl-
edge of IgE epitopes on the allergen
surface is essential. Phl p 4 is one of the
most important allergens from timothy
grass (Phleum pratense) and thus cause of
the hay fever as well as other diseases that
evoke from pollen allergy. This highly
cross-reactive allergen is recognized by 70
88% of grass pollen allergic patients and
thus should be part of potential recombi-
nant vaccine.
Method: Recombinant Phl p 4 has been
produced in Pichia pastoris. Crystals grew
in sitting drop vapor diffusion system. 3D
structure was determined using X-ray crys-
tallography and reticuline oxidase was used
for molecular replacement. Enzymatic
activity was determined monitoring DCPIP
concentration at a spectrophotometer.
Result: Crystals have been obtained in two
conditions diffracting to 1.9 and 1.65 A.
Structure was solved in both conditions.
Known linear epitopes were mapped on to
the surface of the protein and additional
putative epitopes were predicted using
SPADE epetope prediction tool with help
of cross-reactivity data from related aller-
gens. Protein is a glucose dehydrogenase
when using DCPIP as the electron acceptor.
Poster Session 56 Novel approaches to allergy diagnosis
Conclusion: We have solved the rst group
4 pollen allergen structure. The structure
reveals the fold and nature of the protein,
explains its enzymatic activity and is base
for hypoallergenic recombinant vaccine
development.
1384
Creation of hypoallergenic actinidin, the
major kiwifruit allergen by error-prone
PCR for therapeutic applications
Popovic, M1; Prodanovic, R1; Ostafe, R2; Schillberg, S3;
Fischer, R2; Gavrovic-Jankulovic, M1
1
Faculty of Chemistry, Biochemistry, Belgrade, Serbia;
2
RWTH Aachen University, Institute for Molecular
Biotechnology, Aachen, Germany; 3Fraunhofer Institute
for Molecular Biology and Applied Ecology (IME), Plant
Biotechnology, Aachen, Germany
Background: Standard immunotherapeutic
approaches in food allergy are not ade-
quate due to unacceptably high risk-to-
benet ratio, however hypoallergenic forms
of molecules with reduced IgE binding
potential seem to be promising concept.
Aim: The idea behind the creation of
recombinant allergen library was to modify
the surface topography of mayor kiwifruit
allergen actinidine (Act d1) in a way as to
reduce IgE binding, while retaining the
folding pattern of the a-carbon backbone
thus preserving surface structures capable
of generating an IgG response that would
compete with IgE for binding to the natu-
ral allergen.
Methods: Diversication of the gene
encoding the mature form of the actinidin
was done via error prone PCR, followed
by cloning of the library into the Aga2
yeast surface display expression system.
Expressed mutants were sorted with Fluo-
rescence Activated Cell Sorter (FACS)
based on preferential binding for rabbit
anti actinidin IgG antibodies and dimin-
ished binding to IgE antibodies from sera
of kiwifruit allergic individuals. After three
rounds of sorting by FACS, the IgE bind-
ing of selected mutants was also assessed
by both dot blot and Enzyme linked
immunosorbent assay (ELISA) for their
ability to compete with wild type recombi-
nant actinidin. In vitro reactivity of selected
mutants was assessed by analyzing baso-
phil activation from kiwifruit allergic
patients using ow cytometry. The allergen
threshold sensitivity was determined as
CD63 upregulation on CD203c-identied
basophils.
Results: FACS sorting enabled enrichment
of the library as the ratio of IgE positive /
IgG positive cells decreased from 42% in
the rst round to only 16% in the third
round of sorting. Randomly picked colo-
nies were further analysed by dot blot and
ELISA using pool of six kiwifruit allergic
Allergy 66, Suppl. 94 (2011): 482642  2011 John Wiley & Sons A/S
patients. Two mutants selected from the
sorted library exhibited reduced IgE bind-
ing in both assays compared with wild type
Act d 1. Compared to both natural and
recombinant wild type actinidin one of the
mutants exibited no ability to activate ba-
sophils.
Conclusion: This kind of mutational analy-
sis could be a useful technique in the devel-
opment of vaccines for allergy therapy,
since substitution or deletion of amino
acids within the IgE binding epitopes could
result in reduction or elimination of the
anaphylactic potential of the molecule as
was suggested for recombinant Bet v1.
1385
Generation and epitope analysis of
human monoclonal antibody isotypes
with specificity for the timothy grass
major allergen Phl p 5a
Hecker, J1; Diethers, A1; Etzold, S2; Seismann, H1;
Michel, Y1; Plum, M1; Bredehorst, R1; Blank, S1; Braren,
I3; Spillner, E1
1
University of Hamburg, Institute of Biochemistry and
Molecular Biology, Hamburg, Germany; 2PLS-Design
GmbH, Hamburg, Germany; 3University Medical Center
Hamburg-Eppendorf, Hamburg Center for Experimental
Therapy Research (HEXT), Hamburg, Germany
Background: The scarcity of monoclonal
human IgE antibodies with specicity for
dened allergens is a bottleneck for the
molecular characterization of allergens and
their epitopes. Insights into the characteris-
tics of such antibodies may allow for anal-
yses of the molecular basis underlying
allergenicity, allergen cross-reactivities,
standardisation of allergens as well as
improvement of allergy diagnostics and
therapeutics. Here we report the generation
and application of the rst set of authentic
human IgG, IgE and IgA antibodies.
Method: On the basis of a Phl p 5a specic
antibody fragment, a lambda light chain
and the IgG1, IgG4, IgE, IgA1, and IgA2
heavy chains, the corresponding human
immunoglobulins were constructed and
produced in mammalian cells. In parallel, a
murine hybridoma line with specicity for
Phl p 5a was established, recloned and pro-
duced as human chimeric IgE. Specicity
and immunoreactivity of the resulting anti-
bodies were assessed applying different
immunological and cellular assays.
Result: All Phl p 5a-specic antibody iso-
types were successfully cloned and pro-
duced in mammalian cells. After
purication, specicity of the antibodies as
well as their immunoreactivity to the aller-
gen was assessed. Applicability in allergy
diagnostics was conrmed by establishment
of articial human sera. Functionality of
both antibodies was further demonstrated
in receptor binding studies and mediator
release assays using humanized rat baso-
phil leukaemia cells (RBL-SX38) suggest-
ing the presence of spatially separate
epitopes. By using Phl p 5 fusion proteins
and recombinant IgE in immunoblotting
and mediator release assays we assigned
the epitope of the authentic IgE to a pre-
dominantly looped stretch exclusively
available in Phl p 5a.
Conclusion: In summary, the Phl p 5-spe-
cic antibodies are the rst full set of their
kind and represent valuable tools for stud-
ies of fundamental mechanisms and struc-
ture/function relationships of allergens
with the humoral immune response.
1386
Higher prevalence of IgE to Mal d 1 and
Pru p 1 in PR-10 patients with clinically
relevant food allergy
Scala, E; Zennaro, D; Ferrara, R; Bernardi, M;
Alessandri, C; Santoro, M; Palazzo, P; Pomponi, D; Liso,
M; Mari, A
IDI-IRCCS, Center for Molecular Allergology, Rome,
Italy
Background: Allergy to plant-derived foods
has been described most frequently as a
pollen-related phenomenon. Bet v 1-homol-
ogous allergens belong to the pathogenesis-
related proteins (PR-10) and constitute a
group of closely related molecules found in
pollen (e.g. birch, hazel and oak) and
plant-derived food (e.g. Rosaceae, Umbel-
liferae). Bet v 1-homologous structures are
very labile, and as a result they almost
exclusively induce the so-called oral allergy
syndrome, unless severe reactions have
been seldom reported.
Method: We selected Bet v 1 allergic indi-
viduals as assessed by means of the Immu-
noCAP System (Phadia, Sweden).
Demographic and clinical information were
recorded by means of the InterAll software
version 3.0 (Allergy Data Laboratories s.c.,
Italy). The ISAC103 allergen microarray
(PMD, Austria) has been used to test spe-
cic IgE reactivity to a panel of PR-10 mole-
cules, namely Act d 11, Aln g 1.0101, Api g
1.0101, Ara h 8.0101, Bet v 1.0101, Cor a
1.0101, Cor a 1.0401, Dau c 1.0101, Gly m
4, Mal d 1.01081, Pru p 1.0101.
Result: Ninety-ve allergic individuals (55
female, 59.8%), showing specic IgE levels
to Bet v 1 above 1 kUa/l, up to 100 kUa/l,
represented our study group. Fifty-three
patients complained about symptoms after
fruit ingestion, 42 patients had no problem
with plant derived food. We sought to dis-
criminate between patients with food
allergy from those being tolerant. CAP Bet
v 1 IgE, using a cutoff value >9.23 kUa/l,
had a sensitivity of 64.2% and a specicity
of 66.7% (70.8% PPV and 59.6% NPV).
When evaluating patients with food allergy
with ISAC, the top ranked molecules in
525

the population studied were Bet v 1.0101
(91.3%), Cor a 1.0101 (88%), Mal d
1.0108 and Pru p 1.0101 (73.9% both).
Patients with food allergy showed signi-
cantly higher prevalence of Aln g 1.0101,
Api g 1.0101, Ara h 8.0101, Cor a 1.0101,
Cor a 1.0401, Mal d 1.0108 and Pru p
1.0101 IgE recognition. Mal d 1.0108 and
Pru p 1.0101 evaluation (cutoff
value > 0.1 and > 1.08 kUa/l, respec-
tively) showed a higher prognostic ef-
ciency than simple Bet v 1 measurement.
Conclusion: Although representing the
molecule recognising most of the patients,
Bet v 1 itself cannot be useful in discrimi-
nating patients with associated food
allergy. The combined use of allergenic
molecules spotted on the ISAC microarray
is useful in establishing an appropriate
clinical diagnosis in PR-10 sensitized
subjects.
1387
Purification of basogranulin, a unique
product of human basophil granules
Fawzy, A1; Nicholas, B2; Schramm, G3; Haas, H3; Walls,
A4
1
University of Southampton, Southampton General
Hospital/National Research Center of Egypt,
Immunopharmacology Group, Southampton/Cairo-
Egypt, United Kingdom; 2University of Southampton,
Southampton General Hospital, Immunopharmacology
Group, Southampton, United Kingdom; 3Division of
Cellular Allergology, Research Center Borstel, Borstel,
Germany; 4University of Southampton, Southampton
General Hospital, Immunopharmacology Group,
Southampton, United Kingdom
Background: The basophil has long been
implicated in the pathogenesis of allergic
disease, but understanding of its precise
contribution has been hampered by a lack
of cell-specic markers. Basogranulin, a
unique constituent of the granules has been
identied using basophil-specic monoclo-
nal antibody BB1, but little is known of its
properties.
Method: Peripheral blood basophils were
puried by negative selection with mag-
netic beads, and lysates examined by SDS
polyacrylamide gel electrophoresis
(PAGE), blue native PAGE or a novel
composite PAGE technique with western
blotting. Components were separated by
gel ltration and cDNA libraries for ba-
sophils were screened. Basogranulin was
detected using immunoblotting procedures
with BB1; and a new monoclonal antibody
(BB5) prepared against puried basogranu-
lin.
Result: Basophils were isolated from
peripheral blood with a purity greater than
87%. Basogranulin was detected by immu-
nocytochemistry in the granules of 80% of
basophils. Immunoscreening of basophil
cDNA libraries with BB1 antibody, failed
to identify positive clones, raising the pos-
526
Poster Session 56 Novel approaches to allergy diagnosis
sibility that BB1 reacts with a complex
rather than a single peptide, or that the
cDNA for basogranulin is too large to be
represented in the library. Western blotting
of basophil lysates following SDS PAGE
revealed BB1 binding to a band at the top
of the separating gel (>250 kDa). Treat-
ment of basogranulin with reducing agents
(dithiothreitol or b-mercaptoethanol) led to
complete loss of antibody binding. On blue
native PAGE, there was a major band at
3 MDa and a minor band too big to enter
the gel. Application of a novel composite
gel procedure allowed better resolution of
the major band. On gel ltration of baso-
phil lysates, BB1 binding on dot blotting
was restricted to a fraction that eluted
close to the void volume. Cetyl pyridinium
chloride (CPC) precipitation of glycosami-
noglycans led to a reduction in BB1 bind-
ing, as did treatment with pronase (but not
hyaluronidase). BB1 binding was enhanced
with deoxycholic acid and SDS, but
reduced with NP-40 and Triton X-100
detergents. Basogranulin was puried from
basophil lysates using heparin agarose,
carboxymethyl cellulose and butyl agarose
chromatography, and a single band of
approximately 60 kDa detected on western
blotting with BB1. BB5 detected a compo-
nent of the same size on western blots.
Conclusion: Basogranulin consists of a
macromolecular complex of proteins (with
S-S bonds) and glycosaminoglycans.
1388
Identification and characterisation of a
basic polygalacturonase as a major
Cupressus sempervirens pollen allergen
Shahali, Y1; Sutra, J1; Haddad, I2; Vinh, J2; Mari, A3;
Chollet-Martin, S4; Charpin, D5; Senechal, H1; Poncet, P1
1
ESPCI-ParisTech, LSABM UMR 7195 CNRS, Paris,
France; 2ESPCI-ParisTech, SMBP, USR 3149, Paris,
France; 3CMA, Rome, Italy; 4INSERM U996, Unity of
Immunology Autoimmunity and Hypersensibility,
Bichat-Claude Bernard Hospital, Paris, France; 5North
Hospital, Marseille, France
Background: Although polygalacturonases
(PGs) represent major allergens in several
Cupressaceae pollen grains, no protein
from this family has been reported and
identied so far in Italian cypress (C. sem-
pervirens, Cups) pollen. The aim of this
survey was to investigate, by using a prote-
omic approach, the presence of an allergen
homologous to already characterized Cu-
pressaceae PGs in Cups pollen.
Method: Water and PBS pollen extracts
were resolved by two-dimensional gel elec-
trophoresis (2-DE) and assayed with sera
of allergic subjects. The IgE binding pro-
teins were identied by MALDI-TOF mass
spectrometry analysis. The prevalence of
IgE sensitization to main Cups pollen aller-
Allergy 66, Suppl. 94 (2011): 482642  2011 John Wiley & Sons A/S
gens was obtained by means of double one
dimensional gel electrophoresis (D 1-DE).
Result: PBS pollen extracts of Cups pollen
contained two water-unextractable aller-
genic protein spots homologous to cypress-
cedar PGs. These polypeptides exhibited
the same pI (99.5) and different molecular
masses of 43 and 63 kDa. D 1-DE analysis
followed by immunoblotting demonstrated
that the prevalence of IgE sensitization to
this basic allergen reaches 55% of the stud-
ied cypress allergic patients (n = 30).
Conclusion: The 4345 kDa allergenic frac-
tion of Cups pollen includes two major
allergens: Cup s 1 and a basic polygalactu-
ronase. The intensity of IgE binding to this
basic allergen substantially varied accord-
ing to the allergic individuals. For some
tested allergic patients (17%), this protein
seems to be more immunodominant than
other cypress pollen allergens.
1389
Comparison of in vivo allergenic activity
of recombinant grass pollen allergens
(Phl p 1, Phl p s, Phl p 5, Phl p 6) and a
hybrid molecule consisting of their
epitopes
Douladiris, N1; Piskou, K1; Savvatianos, S1; Linhart, B2;
Vrtala, S2; Sifneos, M1; Valenta, R2; Papadopoulos, N1
1
University of Athens, Allergy Research Center, Athens,
Greece; 2Div. of Immunopathology, Dept of
Pathophysiology, Medical University of Vienna, Vienna,
Austria
Background: The diagnostic approach to
grass pollen allergy may become more
detailed and efcient by detecting specic
IgE to its allergenic components. The
objective of our study is to evaluate the in
vivo allergenic activity of recombinant
grass pollen allergens (Phl p1, Phl p2, Phl
p5, Phl p6) and a hybrid molecule of their
epitopes, in grass pollen allergic children.
Methods: Seventy-six children (15 girls, 5
C
15 years) sensitized to grass pollen par-
ticipated in the study. Skin prick tests were
performed with rPhl p1, rPhl p2, rPhl p5,
rPhi p6, the hybrid molecule at a dose of
30 mcg/ml of each protein, timothy and
Bermuda commercial extracts, out of sea-
son. Mean diameters were measured and
compared at diameter 3 mm as a thresh-
old of positivity.
Results: Hybrid molecule detected 61/68,
61/34, compared to timothy and/or Ber-
muda extract and to a combination Phl p
1/5 SPTs respectively. Using ROC analysis,
in our population we observed a Phl p 1
C 5 C 2 C 6 tendency for those with
Bermuda>Timothy (SPTs), while a Phl p
5 C 1 C 2 C 6 tendency shown for
those with Timothy>Bermuda (SPTs).
Non-detected cases were under the positiv-
ity threshold as settled.
Poster Session 56 Novel approaches to allergy diagnosis
Conclusion: A single recombinant hybrid
molecule consisting of the four major Phl
p allergens at a concentration of 30 mcg/
ml was able to detect the majority of grass
pollen sensitized children.
1390
High potential of cross-reactive carbohy-
drate determinant -reduced plants for
improving routine allergy diagnosis
Mertens, M1; Kaulfurst-Soboll, H2; Brehler, R1; von
Schaewen, A2
1
Department of Dermatology, University of Munster,
Munster, Germany; 2Institute of Plant Biology and
Biotechnology, University of Munster, Munster,
Germany
Background: A longstanding debate in
allergy is whether or not specic immuno-
globulin-E antibodies (sIgE) against cross-
reactive carbohydrate determinants (CCDs)
are able to elicit clinical symptoms. In pol-
len, food and insect venom allergy, 20% of
patients display in vitro CCD reactivity.
Since the allergenicity of tomato glycoaller-
gen Lyc e 2 was ascribed to glycan chains
alone, we aimed at evaluating clinical rele-
Allergy 66, Suppl. 94 (2011): 482642  2011 John Wiley & Sons A/S
vance of CCD-reduced foodstuff in
patients with CCD-sIgE.
Method: Gene-modied tomato and/or
potato plants with stable reduction of
CCD epitopes were obtained via RNA
interference by silencing N-acetylglucos-
aminyltransferase I (GNTI), an enzyme,
that catalyses the initial step of CCD for-
mation in the Golgi apparatus.
Two CCD-positive patient groups were
analysed: one with tomato and/or potato
food allergy, and another with insect
venom allergy reporting no symptoms to
potato and/or tomato. The latter served as
control for only CCD-dependent reactions
in immunoblot, basophil activation, and
streptavidin-based in-house-made Immuno-
CAP analyses. Furthermore, non-allergic
and CCD-negative food-allergic patients
were investigated.
Result: Successful CCD reduction in
tomato and potato plants was conrmed
by immunoblot analyses using a CCD-spe-
cic rabbit serum and CCD-positive
patient sera. Basophil activation tests
(BAT) with CCD-positive patients showed
decreased (re)activity with CCD-reduced
foodstuff, most obvious in the insect
venom-allergic but less in the food-allergic
group, indicating that in vivo reactivity is
primarily based on peptide- and not CCD-
sIgE. Peptide epitopes remained unaffected
in CCD-reduced plants, which was veried
with CCD-negative patients displaying sim-
ilar reactivity with wild-type and CCD-
reduced plant extracts in immunoblot anal-
yses and BATs. In-house-made Immuno-
CAPs, applied to investigate feasibility in
routine diagnosis, conrmed the results of
the BAT at the sIgE level.
Conclusion: Although indicating no allergic
symptoms to tomato and potato, CCD-
positive insect venom-allergic patients
showed a positive BAT with wild-type
plant-food extracts. This suggests low clini-
cal relevance of CCD-sIgE. Since with
CCD-reduced plant food extracts, the acti-
vation of basophils was strongly decreased,
this proof-of-principle study demonstrates
usefulness of CCD-reduced foodstuff to
minimize false-positive results and high
potential for improving already existing
diagnostic tools (i.e. BAT and sIgE deter-
mination).
527

Allergy diagnosis: what can we learn from new methods?
1391
Basophil activation test and iodinated
contrast medium immediate hypersensi-
tivity
Rouzaire, P1; Foray, A1; Bienvenu, F1; Dubost, R2;
Mullet, C2; Diot, N2; Nosbaum, A3; Dubost, J2; Berard,
F3; Bienvenu, J1
1
Immunology Laboratory, Lyon-Sud University
Hospital, Pierre-Benite, France; 2Allergo-Anesthesia
Unit, Anesthesia department, Lyon-Sud University
Hospital, Pierre-Benite, France; 3Allergology and
Clinical Immunology department, Lyon-Sud University
Hospital, Pierre-Benite, France
Background: In 2009, a European multi-
centric study reported that 50% of immedi-
ate hypersensitivity reactions (IHR) to
iodinated contrast medium (ICM) consisted
in allergic reactions (1). In 2010, another
work demonstrates the excellent negative
predictive value of skin tests (2). The aim
of our study is to evaluate the perfor-
mances of the basophil activation test
(BAT) in the investigation of IHR to ICM.
Method: During the year 2010, 13 patients,
consulting for the exploration of an IHR
to ICM, were addressed to our lab for a
BAT with an ICM.
Result: Among the 13 patients, two pre-
sented positive BATs for the culprit ICM.
These two patients have developped severe
reactions (grade 4 according Ring and
Messmer). The rst patient was positive
for iobitridol and ioxaglic acid in BAT,
whereas iopamidol and iodixanol were neg-
ative. Skin tests were not realizable. The
second patient was positive for ioxaglic
acid, both in BAT and skin tests. In con-
trast, the two tests were negative with iom-
eprol, iobitridol and iodixanol. Among the
11 patients with negative BATs, skin tests
were also negative for eight (concordance:
73%). A reintroduction test under preme-
dication was performed for three of these
eight patients, and was well tolerated. The
three patients with discordant results (neg-
ative BATs and positive skin tests) were
considered as allergic, and were not rein-
troduced.
Conclusion: BAT seems thus to be a good
diagnostic tool in the exploration of
patients who have presented IHR to ICM,
in complement to skin tests. BAT is also of
particular interest in cases where skin tests
are not available or interpretable. Larger
surveys are still needed to conrm these
preliminary data.
528
Poster Session 57
1. Brockow K, Romano A, Aberer W, et
al. Skin testing in patients with hyper-
sensitivity reactions to iodinated con-
trast media: a European multicenter
study. Allergy. 2009; 64:234241.
2. Caimmi S, Benyahia B, Suau D, et al.
Clinical value of negative skin tests to
iodinated contrast media. Clin Exp
Allergy. 2010; 40:805810.
1392
Impulse oscillometry: an alternative
method to conventional spirometry in
asthmatic children
Tufekci, S1; Arikoglu, T1; Helvaci, I2; Kuyucu, S1
1 73%; P < 0.05). The highest performance
Pediatric Allergy and Immunology Department, Mersin
University Hospital, Mersin, Turkey; 2Biostatistics and
Bioinformatics Department, Mersin University Mersin,
Turkey
Background: The impulse oscillometry
(IOS) is a technique to assess airow
obstruction in patients not able to perform
forced breathing maneuvers. However, the
value of IOS technique in the routine eval-
uation of asthmatic children is still
debated. The aim of this study was to eval-
uate the relationship between impulse os-
cillometry and the gold standard
spirometry parameters in asthmatic chil-
dren.
Methods: 107 stable asthmatic children
aged 717 years performed rst IOS and
then spirometry, both before and after
salbutamol inhalation, on the same occa-
sion. IOS parameters were airway resis-
tance at 535 Hz (R 535), airway
reactance at 535 Hz (X 535), airway
impedance (Zr), resonant frequency (Fres)
and reactance area (XA), and spirometric
parameters were forced vital capacity
(FVC), forced expiratory volume in 1 s
(FEV1), FEV1/FVC ratio, peak expiratory
ow (PEF) and maximum mid-expiratory
ow (MMEF). Correlations between raw
values of IOS parameters and percent pre-
dicted values of spirometry parameters
were calculated by Pearsons correlation
coefcients. The discriminating perfor-
mances of IOS parameters were determined
by receiver operating curves (ROC).
Results: Coefcients of correlations were
poor but signicant when percent predicted
values of spirometry and raw values of
impulse oscillometry were compared. The
Allergy 66, Suppl. 94 (2011): 482642  2011 John Wiley & Sons A/S
following parameters were found to be sig-
nicantly correlated (r = 0.250.50,
P < 0.05): FVC with X5; FEV1 with Zr,
X5-25, XA, and Fres; FEV1/FVC with
X5-25, XA, and Fres; PEF with Zr, R5,
R10, X5-20, XA, and Fres and MMEF
with Zr, R5-15, X5-25, XA and Fres.
When the distinctive performance of the
raw values of IOS parameters were evalu-
ated according to the predened spiromet-
ric cut-off values by using ROC analysis,
high and statistically signicant area under
curve values (AUC) were obtained from
XA, X15 and R5 for PEF (AUC = 61
63%; P < 0.05) and R5-15, R35, X5-20,
Zr, XA and Fres for MMEF (AUC = 62
was found with the R5 (post R5 minus pre
R5 /pre R5) value (AUC = 60%,
P = 0.089, cut off value )16.67),
although not statistically signicant, to
detect reversibility after bronchodilator
inhalation.
Conclusion: IOS measurements signicantly
correlate with spirometry parameters, espe-
cially with those reecting small airway
disease in asthmatic children. IOS can be
used to demonstrate airow obstruction
when spirometry can not be performed,
especially in preschool age group.
1393
Monitoring latex immunotherapy efficacy
with in vivo and in vitro methods (baso-
phil activation test)
Rodrguez-Trabado, A1; Camara-Hijon, C2; Jimenez-
Timon, S3; Romero-Chala, S2; Garca-Trujillo, J2;
Hernandez-Arbeiza, F3; Ferandez-Pereira, L2
1
Campo Aranuelo Hospital, Allergy Department,
Navalmoral de la Mata, Spain; 2San Pedro de Alcantara
Hospital, Immunology Department, Caceres, Spain;
3
Caceres Hospital, Allergy Department, Caceres, Spain
Background: We report a latex allergy case
with and early improvement after two
months of latex immunotherapy. The
patient was a 47-year-old woman, working
as clinical support, with acute urticaria due
to latex allergy and latex-fruit syndrome
(oral ulcerations and pyrosis with pineap-
ple and kiwi). She had rhinoconjunctivitis
and seasonal asthma with improvement
after immunotherapy with grass pollen.
Two years ago, she developed moderate
persistent asthma, without improvement
after immunotherapy with parietaria
Poster Session 57 Allergy diagnosis: what can we learn from new methods?
Table 1. For abstract 1393 AUTHOR: Please cite Table 1 in text.
% activated basophils
Stimulus before immunotherapy
Negative control 2.5
Latex (25 ng/ml) not performed
Latex (50 ng/ml) not performed
Latex (100 ng/ml) 9.00
Latex (250 ng/ml) 17.50
Latex (500 ng/ml) 28.10
Latex (1000 ng/ml) 31.50
Latex (2500 ng/ml) 27.40
Latex (5000 ng/ml) 25.89
pollen. Apart from daily intake of salme-
terol/uticasone, latex sublingual immuno-
therapy (LSLIT) was prescribed
(maintenance dose: ve drops a day, 5 days
per week). Two months after beginning
immunotherapy, the patient reported toler-
ance to kiwi intake, unknown to pineapple,
but not clear clinical improvement of
asthma.
Method: Prick test, specic IgE and glove-
use test were performed to diagnose latex
allergy. The efcacy of immunotherapy
was carried out by globe-use test and
Basophil Activacion Test (BAT), both per-
formed before beginning LSLIT and two
months later. BAT was performed in whole
heparinized blood. Basohpil population
was detected with CD203-FITC and baso-
phil activation with CD63-PE. Serum sal-
ine was used as negative control and the
bacterian peptide fMLP as positive. A
powder latex extract was tested at different
concentrations (505000 ng/ml). An activa-
tion of at least 15% of basophils after
latex stimulation was required to consider
a result as positive.
Result: Prick test and prick-prick with
latex glove were negative. Total IgE:
82.1 mU/ml. Specic IgE (kU/l): latex
4.28, rHev b 6.01 1.58, negative for rHev
b1, 3, 5, 8 and 11. Before treatment:
Glove-use test: Positive (two habonose les-
sons in hand). BAT was positive with
250 ng/ml and higher concentrations. After
2 months treatment: Glove-use test: Nega-
tive (No lessons after 40 min glove con-
tact). BAT: The treatment increased the
activation threshold (minimum antigen
concentration required for a test positive)
from 250 ng/ml to 1000 ng/ml.
Conclusion: An early improvement in cuta-
neous symptoms and latex fruit-syndrome
is possible with LSLIT. A decrease in the
basophil sensitivity, according to an
improvement in the glove-use test, indi-
cates that BAT can be useful as a non-
invasive test to monitor clinical efcacy of
sublingual immunotherapy with latex.
Allergy 66, Suppl. 94 (2011): 482642  2011 John Wiley & Sons A/S
% activated basophils
after immunotherapy
6.10
4.70
4.30
4.70
7.10
8.50
22.40
15.60
19.40
1394
Development of a enzyme-linked
immunosorbent assay kit for
determination of the major allergen from
Dermatophagoides pteronyssinus, Der
p1, house dust mite
van Oeveren, W1; ten Brinke, O2; van der Graaf, K2
1
Haemoscan, R&D, Groningen, Netherlands; 2Citeq,
R&D, Groningen, Netherlands
Background: Dermatophagoides pteronyssi-
nus 1 (Der p1) is a major mite allergen in
Dermatophagoides pteronyssinus house dust
mite. Currently there are no reliable ready-
to-use-kits for the determination of Der
p1. Therefore we developed a ready-to-use-
ELISA for Der p1.
Method: A ninety-six-wells plate (NUNC
Maxisorp) was coated with a monoclonal
antibody (clone 9-5-1) against Der p1 gen-
erated in a mouse. The monoclonal-anti-
body was obtained from a hybrid between
BALB/c spleen cells and a myeloma cell
line. Samples and a standard are incubated
for 1 h at room temperature. For the stan-
dard we used a semi-puried standard
obtained from Der p1 whole culture mate-
rial. Next, a rabbit-polyclonal antibody
(from rabbits immunized with Der p1) was
used and incubated for 30 min, followed
by a peroxidase-labeled detection antibody
and substrate for detection. Afterwards
cleavage of the substrate was quantied by
measuring the optical density at 450 nm
using a microplate reader. A pre-coated
plate and all other components are
included in the kit.
Result: The working range of the Der p1
ELISA was between 1.5 ng/ml and 21 ng/
ml. The Lowest Limit of Quantication
(LLOQ) was 1 ng/ml. The CVs of the
accuracy and precision were lower than
15%. The ELISA kit was also tested on
robustness, specicity, sensitivity, repeat-
ability and stability. All of these variables
were tested according the EMEA guide-
lines for validation. The ELISA kit shows
a good result for each of these variables.
There is a small cross-reactivity (5%)
against Euroglyphus maynei.
Conclusion: This ELISA kit is useful for
analyzing unknown samples for Dermato-
phagoides pteronyssinus mite allergens and
shows 5% cross-reactivity only with Eu-
roglyphus maynei due to genetic similarity.
There is no other cross-reactivity with any
other mite species for example Der f1, Der
p2, Acarus siro.
1395
Detection of IgG and IgE reactivity to the
major bullous pemphigoid autoantigen
using a microarray system
Pomponi, D1; Di Zenzo, G2; Zennaro, D1; Calabresi, V2;
Hertl, M3; Scala, E1; Bernardi, M1; Mari, A1
1
IDI-IRCCS, Center for Molecular Allergology, Rome,
Italy; 2IDI-IRCCS, Molecular and Cell Biology
Laboratory, Rome, Italy; 3Department of Dermatology
and Allergology, Philipps University, Marburg,
Germany
Background: Bullous pemphigoid (BP) is
an autoimmune bullous disease, in which
direct immunouorescence (IF) studies on
perilesional skin reveal linear basement
membrane zone deposits of IgG. Autoanti-
bodies of BP patients react with BP180
and BP230. IgG are detectable by ELISA,
and recently IgE have been shown to play
a role in the disease, often being positive in
the more severe cases. We sought to evalu-
ate the feasibility of detecting IgE and IgG
to BP180 antigen by using a microarray
system.
Method: The BP180 fragment encompass-
ing aa 490562 (NC16A), obtained as a
GST fusion protein, has been spotted on
an experimental version of the ISAC
microarray (PMD, Austria). E. coli GST
(Sigma) has been used for control pur-
poses. The anti-IgE and -IgG, labelled with
two uorochromes, were used in the exper-
iments as supplied by the manufacturer.
Assays were run by mixing the two second-
ary antibodies. Serum samples from 31
subjects affected by BP, already tested for
IgG to BP180 by ELISA, were used. Seven
of 31 BP sera were tested positive for
ELISA-IgE to BP180. A control group was
set in the allergic population seen at CAM.
All subjects in the control group were
tested using the same microarray.
Result: Positive IgG results obtained by
ELISA on the 31 BP patients were repli-
cated by the ISAC IgG testing in 29/31
sera. IgG reactivities were subtracted from
IgG against the GST present in all BP sub-
jects. A serum with positive IF reactivity
and doubtful IgG reactivity by ELISA was
IgG -negative by ISAC, whereas a second
serum with doubtful ELISA reactivity was
IgG reactive by ISAC. 6 of 7 BP sera were
IgE reactive by ELISA, but only weakly
positive by ISAC. In most cases, it was not
possible to reproduce ELISA IgE reactivity
529

by ISAC. Two of the 143 age-matched
allergic control subjects showed IgE reac-
tivity by ISAC, whereas ISAC-IgG were
positive in 19 (13%). The following results
refer to IgE prevalence to ve allergens in
BP patients and in controls, respectively:
Can f 1 0% versus 0.7%, Der p 2 3% ver-
sus 4.2%, Bet v 1 0% versus 10%, Phl p 1
3% versus 6.3%, Pru p 3 3% versus 0.7%,
and Phl p 12 3% versus 0%.
Conclusion: ISAC microarray system is
suitable for detecting IgG autoantibodies
in patients affected by BP. The combined
detection of IgG and IgE requires improve-
ment of the system. The ISAC microarray
testing allows parallel detection of IgE and
IgG towards other antigens helping to
understand patients clinical prole and
giving suggestions for disease immune
mechanisms.
1396
Investigation of the correlation between
itch and inflammation during the early
and late phases of allergic reaction
Okamoto, N; Tanaka, A; Ohmori, K; Matsuda, A;
Matsuda, H
Tokyo University of Agriculture and Technology, Fuchu,
Tokyo, Japan
Background: Itch is dened as an unpleas-
ant sensation that causes the desire or
reex to scratch which is the one of most
serious clinical symptoms in patients with
atopic dermatitis (AD). Inammation is
also common features observed in skin
lesions of patients with AD. Although vari-
ous inammatory cytokines and mediators
released in affected sites may be involved
in itch induction, the exact relation
between allergic inammation and itch
induction remains uncertain. Particularly
in laboratory mice, researchers inject dif-
ferent kinds of pruritogenic reagents
including histamine and substance P to
evaluate the efcacy of anti-AD drugs.
However, it is unclear whether experimen-
tally induced itch represent real itch in AD
or not. We have therefore investigated rela-
tionship between itch and allergic inam-
mation in experimental models with
laboratory mice.
Method: Compound 48/80, histamine, sub-
stance P, and proteinase-activated receptor
2 agonist SLIGRL-NH2 were injected
intracutaneously into the dorsal skin or
subcutaneously into the left ear of ICR
and C57BL/6 mice, respectively. Passive
cutaneous anaphylaxis (PCA) was also
induced with anti-DNP-IgE and its specic
antigen DNP-BSA. Scratching behavior of
individual mice was captured for 24 h and
analyzed by a novel scratch analyzer
SCLABA-Real (Noveltec Inc., Kobe.
Japan). Ear swelling was measured with a
530
Poster Session 57 Allergy diagnosis: what can we learn from new methods?
digimatic micrometer to assess the dgree of
local inammation.
Result: Compound 48/80 and histamine
induced scratching behavior simultaneously
with inammation in ICR mice. However,
in ICR mice injected with substance P and
SLIGRL-NH2, scratching behavior was
induced independently with inammation.
The same phenomenon was observed in
the PCA model. Furthermore, simulta-
neous occurrence of itch and inammation
in C57BL/6 mice was observed only when
histamine was administered. Regarding the
difference between mouse strains, the num-
ber of scratching behavior induced by each
pruritogen in ICR mice is larger than that
in C57BL/6 mice. On the other hand,
inammation in the late phase of allergic Concurrently, cDNA cloning and sequenc-
reaction in C57BL/6 mice was more severe
than that in ICR mice.
Conclusion: The results indicate that itch
and inammation induced by each prurito-
gen are independently regulated and reac-
tivity varies by mouse strains.
1397
Identification of a new allergen from
amaranthus retroflexus pollen, cobala-
min-independent methionine synthase
Tehrani, M1; Sankian, M2; Assarehzadegan, M3; Falak,
R4; Noorbakhsh, R2; Moghadam, M2; Jabbari, F2; Nouri,
H2; Roozbeh Nasiraii, L5; Varasteh, A2
1
Faculty of Medicine, Department of Microbiology and
Immunology, Mazandaran University of Medical
Sciences, Sari, Islamic Republic of Iran; 2Bu-Ali
Research Institute, Department of Immunology, Faculty
of Medicine, Mashhad University of Medical Sciences,
Mashhad, Islamic Republic of Iran; 3Immunology
Department, Faculty of Medicine, Ahvaz Jundishapur
University of Medical Sciences, Ahvaz, Islamic Republic
of Iran; 4Department of Immunology, Faculty of
Medicine, Tehran University of Medical Sciences,
Tehran, Islamic Republic of Iran; 5Department of Food
Science, Islamic Azad University of Nour, Nour, Islamic
Republic of Iran
Background: Cobalamin-independent methi-
onine synthase (MetE) has been previously
reported as an allergen in oilseed rape (Bras-
sica napus) and Russian thistle (Salsola kali)
pollens. MetE from the S. kali pollen has
been previously identied as an 85 kDa pro-
tein which is broken down into two frac-
tions with the apparent molecular weights
(MWs) of 39 and 45 kDa. In this study, we
report MetE as a new allergen from the
redroot pigweed (Amaranthus retroexus)
pollen.
Method: Thirteen adult patients with
allergy to the A. retroexus pollen were
selected according to clinical history and
positive skin prick test. IgE reactivity of
the A. retroexus pollen extract was evalu-
ated by ELISA and immunoblotting.
Based on conserved nucleotide sequences
of MetEs from different plants, the cDNA
of A. retroexus pollen was amplied and
ligated into the PTZ57RT T/A cloning vec-
Allergy 66, Suppl. 94 (2011): 482642  2011 John Wiley & Sons A/S
tor. The recombinant plasmids were then
subjected to sequencing.
Result: Two IgE reactive bands with the
approximate MWs of 39 and 45 kDa were
detected from the immunoblotting of most
of patients sera (62.5% and 81.25%,
respectively). Furthermore, nucleotide
sequencing of the inserted cDNA fragment
revealed an open reading frame of 2271
bp, encoding a 757 amino acid protein.
Sequence alignment revealed that it had an
extensive homology with the family of
MetEs.
Conclusion: The two IgE binding fractions
with the apparent MWs of 45 and 39 kDa
were assumed to be produced by the
breakdown of the 85 kDa protein, MetE.
ing conrmed that the A. retroexus pollen
extract contained the MetE sequence. The
MetE from A. retroexus pollen is thus
suggested as a new allergen. Efforts are
now underway to produce a recombinant
form of this allergen.
1398
Diagnosis of Chenopodium album allergy
with a cocktail of recombinant allergens
as a tool for component-resolved
diagnosis
Nouri, H1; Varasteh, A1; Afsharzadeh, D1; Tehrani, M2;
Roozbeh Nasiraii, L3; Sankian, M1
1
Bu-Ali Research Institute, Department of Immunology,
Faculty of Medicine, Mashhad University of Medical
Sciences, Mashhad, Islamic Republic of Iran; 2Sari
Medical University, Department of Microbiology and
Immunology, Mazandaran University of Medical
Sciences, Sari, Islamic Republic of Iran; 3Department of
Food Science, Islamic Azad University of Nour, Nour,
Islamic Republic of Iran
Background: Chenopodium album pollen is
one of the main sources of pollen allergy
in desert and semi-desert area and contains
three identied allergens. So, the aim of
this study is determination of the diagnos-
tic potential of the C.album recombinant
allergens in an allergenic cocktail form in
comparison with C.album pollen extract.
Method: Diagnostic potential of the aller-
genic cocktail was investigated in thirty-
two individuals with skin prick tests and
obtained results were compared with the
C. album pollen extract. Specic IgE reac-
tivity against pollen extract and the aller-
genic cocktail was determined by ELISA
and western blotting tests. Inhibition
assays were performed by the allergenic
cocktail as an inhibitor.
Result: The exact sensitization prole of
every patient was identied and showed
that 72%, 81% and 46% of allergic
patients had IgE reactivity with rChe a 1,
rChe a 2 and rChe a 3, respectively.
Almost of C. album allergic patients (30/
32) had specic IgE against the allergenic
Poster Session 57 Allergy diagnosis: what can we learn from new methods?
cocktail. In addition, there was a high cor-
relation between IgE levels to the allergenic
cocktail versus pollen extract. The aller-
genic cocktail was able to completely inhi-
bit IgE binding to natural Ch a 1, Che a 2
and Che a 3 in C. album extract. Positive
skin test reactions were visualized in aller-
gic patients that tested by the allergenic
cocktail.
Conclusion: Reliable results which obtained
from this study conrmed that, the aller-
genic cocktail with high diagnostic poten-
tial could be replace with natural C. album
allergen extracts in skin prick test and
serologic tests.
1399
Sensitisation profiles in polysensitised
patients from a restricted geographical
area: further lessons from multiplexed
component resolved diagnosis
Rossi, R1; Melioli, G2; Monasterolo, G3; Harwanegg, C4;
Rossi, L2; Canonica, G5; Passalacqua, G5
1
Azienda Sanitaria Cuneo 1, Rete di Allergologia
Regione Piemonte, Cuneo, Italy; 2Istituto Giannina
Gaslini, Laboratorio Centrale di Analisi, Genoa, Italy;
3
Azienda Sanitaria Cuneo 1, Laboratorio Analisi
Ospedale di Savigliano e Fossano, Cuneo, Italy;
4
Genosense, Genosense, Vienna, Austria; 5University of
Genoa, Allergy and Respiratory Diseases, Genoa, Italy
Background: The micro-array techniques
for the detection of specic IgE have
improved the diagnostic procedures for
allergic diseases. This method also allows
to dene sensitizations from an epidemio-
logical point of view. We studied the sensi-
tisation pattern in a population of
polysensitized patients with respiratory
allergy, living in a restricted geographical
area in the north-western Italy.
Methods: Consecutive patients with
asthma/rhinitis, living in the province of
Cuneo, and having at least two positive
skin prick test for non related aeroallergens
were studied by a microarray (Phadia,
Milan Italy) which allowed to detect spe-
cic IgE against 103 different allergen com-
ponents.
Results: The 70 patients included had spe-
cic IgE towards a mean of 4.3 allergens/
patient (range 212 allergens). Concerning
pollens, 63 (90%) had specic IgE to at
least one genuine grass pollen allergen, 32
(45.7%) had Ole e 1 specic IgE antibod-
ies, although olive tree is not present in the
area. A relevant percentage of sensitization
to mite was found (47.1%). True co-sensi-
tization to grass-pollen allergens/Bet v 1/
Ole e 1 was observed in 15 individuals
(21.4%). Pru p 1, accounted for the higher
percentage of sensitization in 23 patients
(32.85%), four of whom were co-sensitized
to Pru p 3 and/or Art v 3.
Conclusion: A detailed knowledge of the
sensitisation pattern may have relevant
Allergy 66, Suppl. 94 (2011): 482642  2011 John Wiley & Sons A/S
implications for the prescription of specic
immunotherapy. Moreover, sensitization to
PR-10 (or prolin), frequently associated
to oral allergy syndrome, in some cases
could hide the sensitization to LTPs which
are clinically more relevant.
1400
Allergen composition and allergenicity of
North American and European short rag-
weed pollen
Augustin, S; Cromwell, O; Nandy, A
Allergopharma Joachim Ganzer KG, Reinbek, Germany
Background: Pollen from short ragweed
(Ambrosia artemisiifolia) represents one of
the main causes of seasonal allergic rhinitis
and asthma in North America, with the
pectate lyase Amb a 1 constituting the
most important allergen. Recently short
ragweed occurrence has increased in Japan,
Australia and Europe and simultaneously
atopic sensitization against its pollen seems
to have risen in affected areas. The biggest
short ragweed population in Germany has
been reported for the area around Drebkau
in Brandenburg, where pollen counts of
>2000/m3 air were detected in 2009. A
SIT vaccine for treatment of patients suf-
fering from allergy against short ragweed
pollen should be efcient in the treatment
of all affected individuals. The extent of
the similarity between American short rag-
weed and that growing elsewhere is
unknown. In order to investigate whether
the short ragweed now found in Europe is
identical to its progenitor with regard to
allergen composition and allergenicity, pol-
len and plants were collected in the area of
Drebkau and extracts were compared to
those sourced from pollen collected in
North America.
Method: Pollen extracts were separated by
2-D-gelelectrophoresis and compared
regarding their protein composition. IgE
reactivity was analyzed following Western
Blotting. Detected proteins were identied
by mass spectrometry. For additional com-
parison, genomic DNA was extracted from
European short ragweed leaves, Amb a 1
isoallergens amplied by PCR and com-
pared to sequences of known Amb a 1 iso-
allergens from North American plants.
Results: Comparative 2-D-gelelectrophore-
sis demonstrates that the relative protein
composition of European and North
American short ragweed pollen extracts
varies only marginally both qualitatively
and quantitatively. In the two preparations
the isoallergen Amb a 1.01 represents the
most dominant Amb a 1 form. For the
majority of Amb a 1 variants IgE reactiv-
ity correlates with protein abundance.
Amplication of Amb a 1 genes from
European short ragweed conrms sequence
conservation.
Conclusion: European and North Ameri-
can short ragweed pollen are comparable
with regard to identity and relative
amounts of Amb a 1 isoallergens making a
common SIT vaccine for individuals from
both areas feasible.
1401
Proenzyme and active actinidin from ki-
wifruit differs in IgE reactivity
Grozdanovic, M1; Petersen, A2; Lindner, B2;
Atanaskovic-Markovic, M3; Polovic, N1; Gavrovic-
Jankulovic, M4
1
Department of Biochemistry, Faculty of Chemistry,
University of Belgrade, Belgrade, Serbia; 2Borstel
Research Center, Borstel, Germany; 3Faculty of
Medecine, University of Belgrade, Immunology,
Belgrade, Serbia; 4Department of Biochemistry, Faculty
of Chemistry, Belgrade, Serbia
Background: Actinidin (Act d 1) is an
abundant cystein protease and the major
allergen from kiwifruit. Actinidin exists in
the form of a proenzyme and as an active
protease in kiwifruit; however their individ-
ual contribution to the IgE reactivity to
this allergen source has not been evaluated.
Method: The proteolytical activity of the
actinidin preparation, isolated under native
conditions, was determined by a caseinolyt-
ic activity assay, and by zymogram with
gelatin as substrate. N-terminal sequencing
was performed by Edman degradation
(LPSYVD) and the primary structure of
both actinidin forms were conrmed by
mass ngerprint (UniProtKB P00785). IgE
reactivity was determined by means of wes-
tern blotting with allergic patients sera.
Result: Actinidin preparation, isolated
under native conditions, represents a mix-
ture of proenzyme and active enzyme. A
remarkable difference in electrophoretic
mobility of pro- and active actinidin was
observed in SDS-PAGE, both under reduc-
ing and nonreducing conditions. Moreover,
IgE reactivity in Western blot was shown
only for proactinidin while active actinidin
showed no IgE reactivity.
Conclusion: The difference in IgE reactivity
of the two forms of actinidin show the
necessity for determining the amount of
proenzyme and active actinidin in kiwifruit
extract preparations intended for diagnos-
tic purposes, especially for the evaluation
of the prevalence of Act d 1 positive
responses in prospective kiwifruit allergic
patients.
531

1402
Augmented telomerase activity and cyto-
kines in TH cells of parthenium dermati-
tis patients
Anand, V1; Akhtar, N1; Rai, M1; Sharma, A2
1
All India Institute of Medical Sciences, Department of
Biochemistry, New Delhi, India; 2All India Institute of
Medical Sciences, Biochemistry, New Delhi, India
Background: Air-borne contact dermatitis
(skin allergy) is a chronic health problem
in India. It has very little mortality but dis-
ease normally continues to persist with var-
iable remission and relapses causing
distress and morbidity. A cosmopolitan
weed, Parthenium hysterophorus is the com-
monest cause of the parthenium induced
air-borne contact dermatitis. It is a cell-
mediated immune injury to the skin caused
by activated TH lymphocytes in patients
sensitized to the weed Parthenium. Earlier,
it was considered that somatic cells do not
express telomerase activity. Recently, telo-
merase activity in T cells of peripheral
blood has been demonstrated. The aim of
study is to demonstrate telomerase activity
and cytokines of TH cells in peripheral
blood of Parthenium dermatitis patients.
Method: Thirty patients (Positive on
patch-test) and 30 healthy controls were
included in this study. The blood was
drawn from the patients after patch test
(using Van der Bend square chambers) on
fourth day prior to any immunosuppres-
sive therapy. Serum levels of cytokines
(IL-2 and IFN-c) were estimated by high
sensitivity ELISA. PBMC was separated
from blood by histopaque density gradient
centrifugation. TH cells were isolated from
PBMC by Miltenyi magnetic cell sorter.
Telomerase activity was determined by
PCR-ELISA and simultaneously conrmed
by PAGE analysis. All data were analyzed
statistically.
Result: The mean values of TH1 type cyto-
kines IL-2 and IFN-c were increased sig-
nicantly (P < 0.001), as compared to
controls. A signicant increase in telomer-
ase activity (P < 0.04) was also found in
all patients as compared to the healthy
controls. The positive PCR-ELISA signals
were correlated with TRAP ladder and
visualized in PAGE analysis.
Conclusion: Increased cytokine level sug-
gests TH1 type lymphocyte might be
involved in the pathogenesis of skin
lesions. The elevated cytokine and telomer-
ase activity strengthens that TH-cells are
chronically stimulated in Parthenium der-
matitis patients. This correlated activity in
future can be a specic diagnostic marker
in Parthenium dermatitis.
532
Poster Session 57 Allergy diagnosis: what can we learn from new methods?
1403
Identification and characterisation of
digestion-resistant allergens in walnut
and pecan species
Spiric, J1; Knulst, A2; Nordlee, J1; Koppelman, S3;
Taylor, S1; Baumert, J1
1
University of Nebraska-Lincoln, Food Science and
Technology, Lincoln, United States; 2University Medical
Center Utrecht, Department of Dermatology/
Allergology, Utrecht, The Netherlands; 3University of
Nebraska-Lincoln, Food Science and Technology, De
Bilt, The Netherlands
Background: The allergy to walnut is
thought to be the most common among
tree nuts. Allergic reactions to walnut and
pecan can be severe and occasionally life
threatening. There is limited information
available on characterizing the stability of
walnut and pecan allergens. The aim of the
study is to increase the extraction efciency
of soluble walnut and pecan proteins,
examine the digestive stability of walnut
and pecan proteins, and determine if these
allergens play a role in IgE binding and
cross-reactivity between walnut and pecan
allergic individuals.
Method: Various extraction buffers, pH,
and time/temperature combinations were
used to optimize the extraction yield of sol-
uble walnut and pecan soluble proteins.
Protein digestion experiments were carried
out in simulated gastric uid (SGF) and
simulated intestinal uid (SIF) using differ-
ent ratios of protease to substrate. All pro-
tein proles were visualized under reducing
conditions using sodium SDS-PAGE. The
2S albumin proteins from both species
were identied and isolated using Q-TOF-
HPLC-Mass spectroscopy and fast protein
liquid chromatography (FPLC) using a ser-
ies of hydrophobic interaction and ion
exchange chromatography columns. Addi-
tionally, IgE immunoblots and inhibition
blots were used for qualitative assessment
of binding to soluble proteins, and to
determine the cross-reactivity between
allergic individuals.
Result: Increasing salt concentration of the
extraction buffer increased protein solubil-
ity signicantly more than altering buffer
pH. High molecular weight proteins of
both tree nuts are liable to digestion in
SGF and SIF compared to low molecular
weight proteins, especially the 2S albumins.
These digestion-resistant proteins and pep-
tides retain IgE binding despite extensive
proteolytic digestion.
Conclusion: Optimum solubilization of
walnut and pecan protein is dependent on
high salt concentration and could inuence
the results of diagnostic testing if low-salt
buffers are used for preparation of
extracts. The high degree of amino acid
sequence homology of the 2S albumin pro-
teins of walnut and pecan may account for
the cross-reactivity observed between wal-
Allergy 66, Suppl. 94 (2011): 482642  2011 John Wiley & Sons A/S
nut and pecan-allergic individuals. Addi-
tionally, the digestion stability of the 2S
albumins may contribute to allergic sensiti-
zation to walnut and pecan. Identication
of digestion resistant peptides is an impor-
tant factor contributing to assessment on
food protein allergenicity.
1404
Molecular cloning of grape chitinase
Varasteh, A1; Sankian, M2; Falak, R2; Noorbakh, R2;
Assarezadegan, M2; Tehrani, M2; Abolhasani, A2;
Jabbari, F2
1
Mashhad University of Medical Sciences, Immunology
Research Center, Mashhad, Islamic Republic of Iran;
2
Islamic Republic of Iran
Introduction: Grape is a common causative
of oral allergy syndrome in Iran and path-
ogenesis related (PR) proteins such chitin-
ases have been reported as its main IgE
binding proteins. In this study we deter-
mined to produce a recombinant form of
grape class IV chitinase and evaluate its
IgE binding capacity.
Method: Total RNA was puried from
ripen grapes and used for cDNA synthesis.
Grape class IV chitinase sequence was
amplied by RT-PCR method. The PCR
product as well as the puried pET-21
plasmid were both cut by Xho1 and Not1
and fused by T4 ligase. The recombinant
plasmid was selected by ampicillin resis-
tance and transformed into E.coli BL-
21and E.coli Origami. Since the recombi-
nant protein was produced as inclusion
bodies in both bacteria, the insoluble frac-
tion was extracted and solublized in guani-
dine-HCl and following purication by
metal afnity chromatography subjected
into dialyzing and refolding procedures.
The IgE binding capacity of the native
allergen, as well the solublized and
refolded recombinant forms were evaluated
by western blotting.
Results: The study showed that recombi-
nant protein was produced as inclusion
bodies in both bacteria and optimization
of the expression conditions could not lead
to production of a soluble form of this
protein. Although the puried chitinase
was highly reactive with grape allergic
patients sera; none of the produced recom-
binant forms showed signicant IgE reac-
tivity in dot or western blotting assays.
Conclusion: Our study showed that E.coli
strains are not a suitable host for produc-
tion of recombinant forms of three-dimen-
sionally complicated eukaryotic allergens
such as PR proteins. In our point of view,
applications of eukaryotic hosts such as
insect cells are highly recommended for
production of cysteine rich intracellular
proteins.
Poster Session 57 Allergy diagnosis: what can we learn from new methods?
1405
Cloning, expression and purification of
grape thaumatin like protein (TLP) in a
prokaryotic host
Ketabdar, H1; Falak, R2; Sankian, M2; Chamani, J3;
Moghaddam Hoseinabadi, M2; Roozbeh Nasiraie, L4;
Varasteh, A2
1 1
Department of Biology, Islamic Azad University,
Mashhad Branch, Young Researchers Club, Mashhad,
Islamic Republic of Iran; 2Immunobiochemistry Lab,
Immunology Research Center, School of Medicine,
Mashhad University of Medical Sciences, Mashhad,
Islamic Republic of Iran; 3Islamic Azad University of
Mashhad Branch, Department of biology science,
Mashhad, Islamic Republic of Iran; 4Islamic Azad
University of Nour Branch, Department of food science,
Nour, Islamic Republic of Iran
Background: Thaumatin like proteins
(TLPs) are pathogen related proteins and
may play a role in plant innate immunity.
They are accumulated in edible part of
some sweet fruits such as grape, apple,
kiwi, banana, cherry and tomato during
fruits ripening stage and may cause aller-
gic reactions in sensitive individuals. In this
study, we intended to produce the grape
TLP as a recombinant protein, and to eval-
uate its IgE binding properties.
Method: The cDNA of the grape was
amplied by reverse transcriptase polymer-
ase chain reaction (RT-PCR) using degen-
erate primers based on conserved
sequences of TLP family. The PCR prod-
uct and the expression vector (pET-21)
were both cut by Xho I and Not I and
fused by T4 ligase. The constructed recom-
binant plasmid was then transformed into
E.coli BL-21(DE3) and selected by ampicil-
lin resistance. Since the recombinant pro-
tein was produced as inclusion bodies, the
insoluble fraction was extracted and solub-
lized in 6 M guanidine-HCl. Finally, the
recombinant TLP was puried by metal
afnity chromatography and dialyzed
against refolding buffer. The recombinant
protein and puried native allergen were
evaluated for IgE binding ability by
ELISA, western blotting and inhibition
assays.
Result: Although the recombinant 24 kD
TLP was highly expressed in E.coli and
was easily puried, inhibition experiments
showed that, it could not effectively inhibit
the IgE binding properties of natural TLP.
Conclusion: We expressed and puried
grape TLP for the rst time; however, it
seems that the expressed protein was
weakly IgE reactive. This could be due to
incorrect conguration of the disulde
bridges leading to inappropriate protein
folding. This problem could be resolved by
the expression of the grape TLP in a
eukaryotic host.
Allergy 66, Suppl. 94 (2011): 482642  2011 John Wiley & Sons A/S
1406 1407
Molecular diagnosis: a useful tool to opti- The ASP299GLY polymorphism of Toll-
mise patient selection for specific immu- like receptor 4 has a possible role in peri-
notherapy odontal microorganisms as a pathogenic
factor for atherosclerotic lesions
Gonzalez-Cervera, J1; Rodrguez-Domnguez, B2;
Antoln-Amerigo, D2; Henrquez-Santana, A2; Kaidashev, I1; Skochko, O1; Bobrova, N1; Izmaylova, O1;
Ruiz-Hornillos, F2; Angueira-Lapena, M2 DuBuske, L2
Hospital General de Tomelloso, Allergy, Tomelloso, 1
Research Institute for Genetic and Immunological
Spain; 2Spain Grounds of Pathology and Pharmacogenetics,
Ukrainian Medical Stomatological Academy, Poltava,
Ukraine; 2Immunology Research Institute of New
Background: Southern-central Spain is an
England, Gardner, MA, United States
area where grass and olive are the most
relevant pollen allergens. Grass and Olive
Background: Recently, the pathogenic role
principal pollination period is detected in
of periodontal pathogens in atherosclerosis
May and June. Sensitization to both pol-
was shown. Some microorganisms can pen-
lens is common in our area. Selection of
etrate defense barriers of endothelial cells
adequate allergens for specic Immuno-
and blood vessels due to the defects in
therapy in these patients yields a challenge.
innate immunity. The aim of the study is
The aim of this study is to evaluate
to investigate the role of TLR4 polymor-
whether recombinant allergen components
phisms in the susceptibility to periodontal
could be useful to optimize specic immu-
microbes (PM) in atherosclerotic lesions.
notherapy treatment in our area.
Method: Thirty-one autopsies of coronary
Method: Fifty patients, thirty male, were
arteries from patients who died from coro-
studied, age average was 25.6 years old
nary artery disease and 5 autopsies from
(range: 549 years), all of them suffering
healthy individuals who died from acci-
from seasonal rhinoconjunctivitis and
dents were investigated. The presence of
asthma. These patients were sensitized to
PM in atherosclerotic plaques was investi-
both Phleum and olive pollen, performed
gated by multiplex PCR. The TLR 4
by skin prick test (Alk-Abello) and specic
Asp299Gly polymorphism was determined
IgE (Phadia native allergens). We carried
by PCR with specic primers and conse-
out component resolved diagnosis against
quent restriction analysis.
Phleum pratense: (rPhl p 1,r Phl p5 and
Result: The presence of periodontal patho-
rPhl p7/12) and against olive allergen com-
genic microorganisms in atherosclerotic
ponents: nOle e 1 (Immunocap Phadia) for
plaques was observed in 83.9% of patients
all the 50 patients.
with coronary artery disease. Porphyromon-
Result: Positive results to nOle e 1 were
as gingivalis was found in 64.5%, Trepo-
observed in 50 out of 50 patients (100%).
nema denticola in 41.9%, Actinobacillus
On the other hand, rPhl p 1 and rPhl p 5
actinomycetemcomitans in 32.3%, and Bac-
were positive in 41 out of 50 patients (81%).
teroides forsythus and Prevotella intermedia
Patients who resulted negative to rPhlp1
in 12.9% and 6.5% respectively. In 51.6%,
and rPhlp5 were positive to rPhl p 7/12.
two or more species of periodontal micro-
These data allowed avoidance of inappro-
organisms were found. Only in 11.1% of
priate specic immunotherapy treatment in
samples with atherosclerotic plaques had
19% of the patients studied in our area.
these microorganisms in undamaged tis-
Conclusion: Recombinant allergen determi-
sues. The patients who died from coronary
nation has proved to be both suitable and
arteries disease had expression of the
pivotal in terms of optimizing patient selec-
299Gly allele of the TLR4 gene more fre-
tion for specic immunotherapy in our area.
quently than healthy fatalities [(P = 0.04)
Likewise is a useful tool to determinate
OR 2.92 (1.157.41)].
individual sensitization prole and assists in
Conclusion: Periodontal microorganisms
clearing up potential cross-reactivities.
play an important role in the pathogenesis
of atherosclerosis in patients with coronary
artery disease. The presence of the 299Gly
allele of the TLR4 gene increases the risk
of this disease. TLR4 gene polymorphisms
can inuence immune responses as patients
having the polymorphism 299Gly have
impaired reactions to TLR4 ligands.
533

Clinical and laboratory methods in allergy
1408
Immunochemical detection of nonylphe-
nol in water samples
Piven, N; Burakovski, A; Lukhverchyk, L
Institute of Bioorganic Chemistry of Belarus National
Academy of Sciences, Medical Microanalysis
Laboratory, Minsk, Belarus
Background: Nonylphenol (NP) is the end
product of nonionic surfactants degrada-
tion that can adversely affect humans and
animals by disrupting the functioning of
different regulatory units of the organism,
resulting in the development of serious
consequences for living organisms and
environmental objects. Therefore it is nec-
essary to identify and detect the NP, espe-
cially in aquatic environments, food, etc.,
and to assess the risk on human health
under the inuence of NP.
Method: A new express methods of
enzyme-linked immunosorbent assay
(ELISA format) and immunobiosensor
analysis (based on the combined use of the
specicity of immunochemical reaction
antigen-antibody and the phenomenon
of surface plasmon resonance) for the iden-
tication and quantitative detection of NP
in aquatic environments were developed.
Result: The new developed immunochemi-
cal method of NP detection in water sam-
ples (ELISA format), has the following
characteristics: high sensitivity (68 ng/ml);
strict specicity; wide range of determined
concentrations (6400 ng/ml), reliability
and methodological availability for the
practical use. Analysis duration 4 h with-
out taking into account the stage of the
plate preparation. A new method of immu-
nobiosensor determination of NP in water
samples has the following characteristics:
sensitivity of analysis 35 ng/ml (this is
enough for determination of potentially
dangerous NP concentrations for the
organism); working range 101000 ng/
ml; the ability for the repeated utilization
of the biosensor chip. Analysis duration
10 min (without preliminary preparation of
the sensor chip). The comparative analysis
of our two methods on water samples, con-
taining various concentrations of NP,
showed comparable results.
Conclusion: The developed immunochemi-
cal methods for identication and quanti-
tative detection of NP in water samples
can be recommended for water quality
534
Poster Session 58
control before and after its treatment and
monitoring of the contamination of water
objects of environment (drinking, tap, foul
water, water from the reservoirs, etc.), as
well as to research the mechanisms of its
effects on the human body and animals.
1409
Immune responses to 6-month whole
body vibration (acceleration) training in
elderly women: a pilot study
Shirolapov, I1; Pyatin, V1; Zhestkov, A2; Nikitin, O1;
Alekseeva, A1; Merkulova, S1
1
Samara State Medical University, Normal Physiology
Department, Samara, Russian Federation; 2Samara
State Medical University, Clinical Microbiology,
Immunology and Allergology Department, Samara,
Russian Federation
Background: WBV (whole body vibration,
acceleration) training has been recently
shown to develop human physiological
parameters by improving muscle perfor-
mance, nervous function, signicantly
increasing bone mineral density, physiolog-
ical secretion of anabolic hormones,
growth and antiaging factors. WBV train-
ing as a new type of physical exercises
could positively inuence many risk factors
of social diseases (including cardiovascular
disorders, osteoporosis, strokes), however,
the data on the safety and efcacy of WBV
loading to immune function are lacking.
The present pioneering trial aimed at inves-
tigating the cell and humoral immune indi-
ces and expression of lymphocyte
activation markers in elderly women dur-
ing 24-ws WBV exercises as compared to
the control.
Method: Thirty-nine elderly women were
randomly assigned to a WBV training
group (n = 20, aged 65 4 years) or a
control group (n = 19, aged 67 4 years).
WBV group trained three times weekly for
24 weeks performing progressive-based sta-
tic and dynamic physical exercises on accel-
eration platform (Power Plate, frequency
3540 Hz, amplitude 2 mm, 30 min daily),
which evoked reexive muscle contractions.
The control group did not participate in
any training. The immune cells counts were
measured by 2-color-FACS. The humoral
factors of immunity were assessed by
means of ELISA.
Result: Signicant changes in the immune
indices were noted after 12 weeks of WBV
Allergy 66, Suppl. 94 (2011): 482642  2011 John Wiley & Sons A/S
training. It has been revealed that some
immune parameters were decreased and at
once levels of pro-inammatory serum
cytokines were above normal that resulted
from dramatically increased intensity of
the intervention loading. The end point
results showed no signicant effect of
24 weeks WBV exercises on the in vitro
immune function including lymphocyte
phenotype number, serum cytokines (IL-8
and TNFa) and serum immunoglobulins
(A, M, G) levels, CD25 and HLA-DR
expression on lymphocytes in elderly
women.
Conclusion: These pioneering ndings sug-
gest that the progression-based 6-month
whole body vibration (acceleration) train-
ing is not a sufcient stress factor affecting
immune function and does not produce
any signicant postexercise suppression of
immune functions in elderly women. Thus
this intervention may be a benecial tech-
nique to be used extensively in clinical
rehabilitation practice.
1410
Optimizing RNA isolation for exosomal
RNA
Eldh, M; Lotvall, J; Malmhall, C; Ekstrom, K
University of Gothenburg, Internal Medicine, Krefting
Research Centre, Gothenburg, Sweden
Background: Exosomes are small RNA
containing vesicles of endocytic origin,
which participate in cell to cell communica-
tion by transferring exosomal RNA
between cells. Exosomes are released by
many cells and can be found in biological
uids, including plasma and breast milk.
Exosomes differ compared to their donor
cells in RNA, protein and lipid composi-
tion. Many studies describe exosomal
RNA from different cellular origins using
different RNA isolation methods giving
rise to different exosomal RNA size distri-
bution patterns. In some publications, it
seems that exosomes are enriched in small
RNA while in others a more wide size dis-
tribution pattern is seen. The aim of the
current study was to determine whether
variation in exosomal RNA quantities, size
distribution and quality could be a conse-
quence of the RNA extraction method
used and to determine the optimal way to
extract RNA from exosomes.
Poster Session 58 Clinical and laboratory methods in allergy
Method: Exosomes were isolated from a
mast cell line (MC/9) and seven different
RNA extraction methods were evaluated
to determine quality, purity, yield and size
of RNA. The methods evaluated were; one
phenol based method, three combined phe-
nol and column based methods, and three
column based methods. The RNA quality
was determined for cellular RNA by the
Bioanalyzer using RNA integrity numbers
(RIN) and the purity was evaluated spec-
trophotometerically at the absorbance 230,
260 and 280 nm. The RNA yield and size
of exosomal and cellular total and small
RNA was analysed using the Bioanalyzer.
The presence of microRNA was conrmed
by real-time PCR.
Result: All methods evaluated extracted
high quality and purity RNA. However,
the exosomal RNA yield and Bioanalyzer
electropherogram patterns differed sub-
stantially between the isolation approaches.
Conclusion: In the current study we present
a unique comparison of seven different
methods for extraction of exosomal RNA.
As the different isolation methods give
such an extensive variation in exosomal
RNA yield and pattern, it is of great
importance to choose the method carefully
depending on the research question at
hand.
1411
In vivo immunogenicity assay for a house
dust mite allergoid preparation to serve
as quantitative indicator of the product
potency
Marcondes - Rezende, M1; Kerkvliet, E1; Gilmartin, L2;
Hilgers, L3; Opstelten, D1; Koppelman, S1; van der Kleij,
H1
1 1412
HAL Allergy BV, Research and Development, Leiden,
The Netherlands; 2Charles River Laboratories Preclinical
Services, Carrentrila, Ballina, Co. Mayo, Ireland; 3Polder
Products B.V., Ophemert, The Netherlands
Background: A feasibility study was con-
ducted to assess the possibility of developing
an in vivo immunogenicity assay for differ-
ent product strengths of PURETHAL
Mites (PUR-M), an allergen preparation
consisting of a mixture of glutaraldehyde-
modied extracts from Dermatophagoides
pteronyssinus and Dermatophagoides farinae
adsorbed to aluminium hydroxide. For of-
cial use as a potency assay for stability test-
ing, the authorities have determined that the
in vivo model must be able to detect a 50%
decrease in product potency. IgG antibody
titres to the allergoid mixture served as
quantitative potency indicators.
Method: BALB/c mice were immunized
two, three or four times by subcutaneous
injection at a 2-week interval with 100 ll
test item containing 2000 AU (normal
dose) or 8000 AU (high dose) of PUR-M.
Allergy 66, Suppl. 94 (2011): 482642  2011 John Wiley & Sons A/S
Graded product strengths were prepared
by diluting the normal and the high stan-
dard dose either 5-fold or 2-fold in buffer
containing aluminium hydroxide and phe-
nol (matrix), to determine the capacity of
the model to detect decreased product
potencies. Serum was collected one week
after the last injection and analyzed for
IgG antibodies against the allergoid. Titres
were determined by ELISA and expressed
as arbitrary units (AU) by comparison to a
reference of pooled sera from immunized
mice.
Result: PUR-M elicited IgG responses
against the allergoid. Decreasing the dose
5-fold signicantly reduced the geometric
mean titre (GMT) after four injections
(P < 0.001) indicating that a 5-fold reduc-
tion in product potency could be detected
(10 mice per group). With an increased
group number of 20 mice, decreasing the
dose 2-fold, the normal dose showed small
differences in immune responses between
groups after four injections (P < 0.05).
However, the difference in GMTs between
groups for the high dose was not signi-
cant.
Conclusion: The data do not support the
detection of a 2-fold reduction in both
product strengths, even with 20 mice per
group. IgG titres to allergoid elicited by
PUR-M can therefore not be used as a
quantitative potency indicator of this prod-
uct. Without extensive model optimization
and using large group numbers, it is not
possible to use an in vivo immunogenicity
model as a read out for stability testing of
immunotherapy products in time.
Measurement of exhaled nitric oxide:
comparison of two analyzers
Yoon, H1; Kwak, H1; Kim, T1; Sohn, J1; Shin, D1; Park,
S1; Kim, S1; Chang, S2
1
Hanyang University College of Medicine, Department
of Internal Medicine, Seoul, Republic of Korea; 2Sungae
General Hospital, Department of Internal Medicine,
Seoul, Republic of Korea
Background: Measurement of fractional
nitric oxide in exhale air (FeNO) is a mar-
ker of airway inammation and useful in
the management of asthma and other
respiratory diseases. The NIOX MINO, a
new hand held device for the measurement
of FeNO, simple and easy to use in clinical
practice. We compared the performance of
NIOX MINO measurement with the sta-
tionary chemiluminescence analyzer
(NOA280i).
Method: We measured FeNO in 101 adult
subjects (aged 2256 years) with NIOX
MINO and NOA280i consecutively at a
ow rate of 50 ml/sec. Of the enrolled sub-
jects, eight subjects (7.9%) have current
asthmatic symptoms or have been diag-
nosed as asthma. The rst acceptable mea-
surement with NIOX MINO and the mean
of three acceptable measurements with
NOA280i were used for analysis.
Result: There was a signicant correlation
between FeNO concentrations between by
NIOX MINO and NOA280i measurement
(r = 0.908, P < 0.0001). The Bland-Alt-
man plot showed a high degree of agree-
ment between the two devices: the mean
inter-device difference was 3.18 ppb and
the limits of agreement were )7.06 and
13.42 ppb.
Conclusion: FeNO levels measured by
NIOX MINO showed a signicant correla-
tion and a high degree of agreement with
those measured by NOA280i. These nd-
ings suggest that NIOX MINO could give
a reliable index for airway inammation in
clinical practice.
1413
Evaluation of the efficacy of montelukast
in maintaining topic steroid-induced clini-
cal and histological response in adult
eosinophilic esophagitis patients
Gonzalez-Cervera, J1; Lucendo-Villarn, A2; Rodrguez-
Domnguez, B3; Arias-Arias, A3; Yague-Compadre, J3;
Angueira-Lapena, M2
1
Allergy Department, Spain; 2Gastroenterology
department, Spain; 3Spain
Background: Eosinophilic esophagitis
(EoE) is characterized by esophageal eosin-
ophilic inltration, determining structural
changes and dismotility symptoms. EoE is
considered to be an atopic associated
inammatory disorder. Allergies have been
linked to the EoE origin. Systemic corti-
costeroids have been one of the most used
drugs, nevertheless topic steroids have
demonstrated to be equally effective and
the adverse effects are lower. However
other drugs with a better security prole in
the long term have been proposed to for
treating EoE. Montelukast is a selective
antagonist drug which specically blocks
the receptor for leukotriene D4 expressed
on eosinophils. The aim of this study is to
determinate whether or not montelukast
could offer benets in replacing steroids
after the eosinophilic inammation has
been solved.
Method: Eleven adult patients nine male
and two female, were studied, age average
34.91 years old (range: 2061 years), all of
them suffering from eosinophilic esophagi-
tis. The patients had exhibited esophageal
symptoms for a mean period of 42.55
months (range: 1272). These patients were
prospectively studied. Esophageal biopsies
were obtained before and after a 6-month
treatment with uticasone propionate
400 mcg twice a day. After that this treat-
535

ment was stopped and treatment with
montelukast 10 mg/day was instituted. A
new endoscopy was foreseen after a new
three months period. Esophageal symp-
toms were assessed by using a question-
naire (validated for achalasia) before and
after uticasone propionate treatment and
after montelukast therapy.
Result: Eosinophilic density into the
esophageal epithelium and lamina propia
was signicantly reduced after a six
month treatment with topical steroids
(P = 0.003) and increased again after
montelukast therapy, but improving the
basal levels; moreover four out of the ele-
ven patients did not complete the schedule
3-month period of treatment with mont-
elukast for referring serious deterioration
in their symptoms at the second month of
treatment.
Conclusion: Montelukast treatment has not
proved to be effective in maintaining histo-
pathological nor clinical response achieved
by topic steroids in adult patients suffering
from eosinophilic esophagitis.
1414
Impact of laryngopharyngeal and gastro-
esophageal reflux on asthma control and
the predictive value of symptoms and lar-
yngeal findings in children with asthma
Kilic, M1; Ozturk, F1; Kirmemis, O2; Atmaca, S3; Guner,
S1; Caltepe, G2; Sancak, R1; Kalayci, A2
1 1
Ondokuz Mayis University Hospital, Allergy and
Clinical Immunology, Samsun, Turkey; 2Ondokuz Mayis
University Hospital, Gastroenterology and Hepatology,
Samsun, Turkey; 3Ondokuz Mayis University Hospital,
Otolaryngology, Head and Neck surgery, Samsun,
Turkey
Background: Even though the relationship
between gastro-esophageal reux (GER)
and asthma has been extensively investi-
gated, there is not much data about the
association between asthma and laryngo-
pharyngeal reux (LPR) in children. A
diagnosis of LPR may be established by
questioning the symptoms, videolaryngo-
scopic evaluation of larynx or double
probe pH monitoring. The aim of this
study was (1) to determine the incidence of
GER and LPR in children with asthma;
(2) to determine the sensitivity, specicity,
positive and negative predictive value of
reux symptom score and laryngeal nding
score to diagnose LPR and GER in chil-
dren with asthma by comparing the results
of double probe pH monitoring study; (3)
to determine the difference between con-
trolled and uncontrolled asthma in terms
of GER and LPR.
Method: A total of 50 patients (23 girls, 27
boys, mean age 10.8 2.3 years) with
mild to moderate persistent asthma were
included in this study. The patients were
divided in two groups according to the
536
Poster Session 58 Clinical and laboratory methods in allergy
asthma control status as controlled
(n = 27) versus uncontrolled asthma
(n = 23). All patients completed the reux
symptom questionnaire and then they
underwent indirect videolaryngoscopic
examination and 24 h double probe (pha-
ryngeal and distal esophageal) pH moni-
toring study.
Result: The incidences of LPR and GER
were found 70% and 46% in asthma,
respectively. Eighteen (36%) of patients
had both LPR and GER. The reux symp-
tom score and laryngeal nding score were
not useful to predict LPR or GER. We
found that there was no association
between asthma control status and LPR
and GER. Furthermore there was no dif-
ference between atopic and non-atopic
patients in terms of LPR and GER.
Conclusion: Laryngopharyngeal reux is a
more frequent condition than GER and
the reux symptom score and laryngeal
nding score do not seem reliable to diag-
nose LPR and GER in children with
asthma. Uncontrolled and atopic asthma
do not have high risk for LPR or GER.
1415
Optimisation of peanut protein crosslink-
ing by oxidase and transglutaminase
enzymes and the effects on IgE binding
Mihajlovic, L1; Radosavljevic, J1; Nordlund, E2; Smit, J3;
Buchert, J2; Cirkovic Velickovic, T1
Faculty of Chemistry, University of Belgrade, Chair of
biochemistry, Belgrade, Serbia; 2VTT biotechnology,
Espoo, Finland; 3Center for Food Allergy, Utrecht
University, Utrecht, The Netherlands
Background: Enzymatic crosslinking is a
procedure with increasing application in
food and pharmaceutical industries. Peanut
(Arachis hypogea) protein extract was
crosslinked by several enzymes including
laccase and tyrosinases from fungal sources
as well as a microbial transglutaminase, in
order to obtain high molecular mass cross-
links with modied allergological proper-
ties.
Method: The extraction process was opti-
mized in order to simplify the procedure
and allow extensive crosslinking by oxidas-
es and transgulataminases enzymes with
potential use in food processing. Cross-
linked proteins were analyzed by SDS-
PAGE on 7% and 12% gels as well as on
composite 0.5% agarose gels. IgE binding
was determined by immunoblot, ELISA
inhibition and basophil activation experi-
ments in four allergic patients. Incorpora-
tion of peanut allergens, Ara h 1, Ara h 2,
Ara h 3 and Ara h 6 was determined by
immunoblot using specic antibodies pro-
duced in mice and rabbits.
Result: The molecular weight of the cross-
linked proteins varied depending on the
Allergy 66, Suppl. 94 (2011): 482642  2011 John Wiley & Sons A/S
enzymes used, with the largest crosslinks
(obtained by laccase and tyrosinase from
A. bisporus) having molecular weight of
around 5MDa. Immunoblots showed that
certain peanut allergens (i.e. Ara h 1, Ara
h 6) are less prone to crosslinking, with
reaction efciency depending on the
enzyme. ELISA inhibition was used to
quantify IgE binding, showing slightly
increased IC50 values for crosslinked pro-
teins made by laccase, transglutaminase
and A. bisporus tyrosinase, indicating a
decrease in IgE binding in these samples.
All of the crosslinked proteins exhibited
reduced basophil activation in the concen-
tration range of 0.510 lg/ml, with results
showing similar or lower activation than
untreated peanut proteins at other exam-
ined concentrations
Conclusion: The obtained crosslinked pro-
teins have molecular masses on the order
of 35 MDa, with moderate reduction of
IgE binding during the crosslinking pro-
cess. Therefore, enzymatic cross-linking
has potential for use in immunotherapy of
food allergies and creation of hypoaller-
genic and safer food products.
1416
Serum diamino-oxidase activity as a
diagnostic test for histamine intolerance
Silar, M1; Music, E2; Korosec, P2; Kosnik, M2; Rijavec,
M2
1
University Clinic of Respiratory and Allergic Diseases,
Golnik, Slovenia; 2University Clinic of Respiratory and
Allergic Diseases, Lab. for Clinical Immunology and
Molecular Genetics, Golnik, Slovenia
Background: Histamine intolerance is
mainly caused by an imbalance of hista-
mine intake and the capacity for histamine
metabolism and degradation. The main
enzyme for metabolism of ingested hista-
mine is diamine oxidase (DAO). Determi-
nation of DAO activity in serum might be
useful for differential diagnosis of hista-
mine intolerance.
Method: Over the 3.5-year-long period we
have recruited 316 patients with suspicion
of histamine intolerance and excluded food
allergy together with 20 healthy controls.
Serum DAO activity was measured with
Enzyme immunoassay for the quantitative
determination of histamine-degradation
activity by DAO in serum. Twenty patients
with histamine intolerance and highly
reduced initial activity of serum DAO
(<40 HDU/ml) went to a histamine-free
diet and after 612 months of histamine-
free diet all clinical parameters and serum
for determination of DAO activity were
taken again.
Results: We found that DAO activity was
signicantly lower in patients than in
healthy control subjects (P < 0.0001).
Poster Session 58 Clinical and laboratory methods in allergy
Furthermore, 54 patients had highly
reduced activity of DAO (<40 HDU/ml).
The main symptoms involved the skin, gas-
trointestinal tract, respiratory system and
eyes. In all 20 patients after the histamine-
free diet the main clinical symptoms typical
for histamine intolerance have disappeared.
Furthermore, the measured values for
activity of serum DAO have increased sig-
nicantly (P < 0.0001).
Conclusion: We can conclude that determi-
nation of DAO activity in serum is a useful
diagnostic tool, together with detailed his-
tory to differentiate between food allergy
and histamine intolerance. It should be
performed in suspected patients with symp-
toms like headache, tachycardia, urticaria,
pruritus, diarrhea and hypotension, where
food allergy was excluded. Furthermore,
our results showed the benet of hista-
mine-free diet, since after the diet majority
of histamine related symptoms have disap-
peared as well as the DAO activity in
serum has increased.
Allergy 66, Suppl. 94 (2011): 482642  2011 John Wiley & Sons A/S
1417
High hydrostatic pressure processing: a
method having high success potential in
pollen protein extraction
Altuner, E1; Ceter, T1; Karakaya, G2; Filiz, E2; Alpas, H2
1
Kastamonu University, Department of Biology,
Kastamonu, Turkey; 2Middle East Technical University,
Food Engineering Department, Ankara, Turkey
Background: High Hydrostatic Pressure
Processing (HHPP) is a food processing
method, in which food is subjected to the
elevated pressure which is mostly between
100 to 600 MPa. Not only in the food
engineering, but HHPP have also other
application areas, such as extraction of
active ingredients from natural biomateri-
als. Proteins, glycoproteins or even a single
peptide which are present in the pollen
wall and cytoplasm could cause the pollen
allergy. In order to produce skin prick test
kits for pollen allergy, the rst step is the
extraction of these molecules. There are
several methods to extract these molecules,
but HHPP is the one which could have a
great opportunity to be used for this pur-
pose.
Method: In this study, Cedrus atlantica
pollens are subjected to 220 and 330 MPa
for 10 and 30 min. After applying HHPP,
the total amount of proteins which are
allergenic and non-allergenic are measured.
Then, the total amount of proteins is com-
pared with the total protein extracted by
conventional extraction method at a physi-
ological pH value. In addition, the micros-
copy of the pollens has also been studied
to observe their morphological changes
after extraction.
Result: As a result, the total protein
extracted by HHPP is 18.0210 lg/ll at
220 MPa for 10 min, 22.5770 lg/ll at
220 MPa for 30 min, 23.3810 lg/ll at
330 MPa for 10 min and 25.9270 lg/ll at
330 MPa for 30 min, while the total pro-
tein extracted in 24 h by the conventional
extraction method is 1.9460 lg/ll. In addi-
tion to these results, visual pollen deforma-
tion and eruption, pollen wall and surface
damage have also been observed.
Conclusion: Since HHPP could cause some
structural changes such as deformation and
membrane damage, the total protein
extracted by this method has been dramati-
cally increased. However, it is known that
HHPP could cause protein denaturation.
For this reason, further studies should be
conducted to observe, whether a denatur-
ation occurred or not.
537

Autoimmunity: disease mechanisms
1418
Rheumatoid arthritis. Relationship
between pro-inflammatory status and
activity of disease
Ciacli, C
Vasile Goldis West University, Department of
Immunology, Arad, Romania
Background: The aim of this study was to
investigate the concentration of Th1 types
cytokines (IL -1, IL-6, and TNF-alpha) in
the peripheral blood of patients with rheu-
matoid arthritis versus healthy individuals.
Method: Twenty patients with rheumatoid
arthritis and twenty-seven normals were
included. The pro-inammatory cytokines
levels were quantied by using ELISA
method (Endogen, Woburg, MA, USA).
Result: Peripheral blood concentrations of
Th1 types cytokines (IL -1, IL-6, and
TNF-alpha) were higher in rheumatoid
arthritis than martors.
Conclusion: This study has detected higher
levels of pro-inammatory cytokines in
arthritis, so reecting the pro-inammatory
status in this diseases, and the increase in
pro-inammatory cytokines serum level is
strongly positive related to the activity
of diseases in patients with rheumatoid
arthritis.
1419
The role of receptor activator of
NF-kappa B ligand in appearance
systemic osteoporosis in psoriatic and
rheumatoid arthritis
Ciacli, C
Vasile Goldis West University, Department of
Immunology, Arad, Romania
Background: The aim of this study was to
investigate the comparative evaluation of
serum levels of NF-kappa B ligand
(RANKL) with bone mineral density values
to identify useful correlations in early diag-
nosis of complications and identify patients
who may benet from specic treatment.
Method: Twenty-seven patients with psori-
atic arthritis, twenty-one patients with
rheumatoid arthritis and twenty normals
were included. The RANKL levels were
quantied by using ELISA method (Endo-
gen, Woburg, MA, USA) and the bone
with dual-energy X-ray absorptiometry
(DEXA).
538
Poster Session 59
Result: Our study demonstrates that there
is a strong negative correlation between
blood values of RANKL and T score val-
ues in patients with psoriatic and those
with rheumatoid arthritis.
Conclusion: Personal contribution of our
study on this topic is to demonstrate the
occurrence of systemic osteoporosis in
patients with psoriatic and rheumatoid
arthritis, complication that is strongly
dependent on the blood values of RANKL
and should be routinely monitored in these
patients to prevent osteoporotic bone frac-
tures, complications that require increased
nancial and social costs. Key words: pso-
riatic arthritis, rheumatoid arthritis,
RANKL, DEXA, systemic osteoporosis
1420
The role of cytotoxic T-lymphocytes in
the immunopathogenesis of rheumatoid
arthritis
Khasanov, U; Sizyakina, L
increased, which ensures their protective
Rostov State Medical University, Department of Clinical
Immunology and Allergology, Rostov-on-Don, Russian
Federation
Background: According to modern ideas,
rheumatoid arthritis (RA) is a heteroge-
neous disease, the basis of the pathogenesis
of which is a complex, poorly understood
combination of genetically determined and
acquired defects in immunoregulatory
mechanisms. The main feature of RA is
the development of abnormal activation of
the immune system in response to potential
pathogens, and sometimes physiologic
stimuli. However, the intimate mechanisms
Islamic Republic of Iran
of RA arent studied enough until now.
Particulary, the role of the cytotoxic level
of immune system in the pathogenesis of
RA is debatable.
The background of the study was to
study the quantitative and functional
parameters of cytotoxic lymphocytes in
seropositive variant of RA with moderate
degree of activity of autoimmune process.
The method used observation of 24
patients with articular variant of seroposi-
tive RA with a moderate degree of inam-
matory activity. 10 healthy donors were
used as a control group. We investigated
the amount ofCD8+cytotoxic lympho-
cytes. The functional capacity of cytotoxic
lymphocytes was determined by intracellu-
Allergy 66, Suppl. 94 (2011): 482642  2011 John Wiley & Sons A/S
lar content of the main effector molecule
granzyme B.
Results: In patients with moderately active
RA, articular variant of seropositive for
rheumatoid factor showed a statistically
signicant decrease of the amount of
CD8+ lymphocytes compared with the
control group (CD8 + 18.30 0.90%
and CD8 + 24 2% in controls). The
data indicate that a moderate degree of
activity of RA is characterized by a
decrease in the amount of cytotoxic lym-
phocytes, but their functional activity is
debatable. The functional activity of cyto-
toxic lymphocytes revealed a statistically
signicant increase in the content of effec-
tor cells wihich produce granzyme B
(CD8 + / granzyme + 27.90 2.00%, in
control of 22.75 1.40%).
Conclusion: Thus, despite the fact that a
moderate degree of activity of RA is char-
acterized by the decrease of the number of
cytotoxic lymphocytes, but functional
activity of lymphocytes signicantly
role in the degree of the activity of the
pathological process.
1421
Evaluation of apoptosis-related genes;
Fas (CD95) and FasL (CD178) polymor-
phisms in Iranian rheumatoid arthritis
patients
Tahoor, M1; Mohammadzadeh, A1; Akhlaghi, M2;
Pourfathollah, A1
1
Tarbiat Modares University, Immunology, Tehran,
Islamic Republic of Iran; 2Tehran university of Medical
Sciences, Rheumatology Research Center, Tehran,
Background: Rheumatoid arthritis (RA) is
an inammatory arthritidis which effect
synoviocytes. The mechanisms that initiate
chronic inammation have not been well
elucidated, but several evidences indicate
that inadequate apoptosis is one of the
chief reasons. Apoptosis signals are essen-
tial for establishing of homeostasis and
adequate immune response dysregulation
of apoptosisrelated genes in the immune
system which could be due to gene poly-
morphisms conduct to autoimmune dis-
eases including rheumatoid arthritis.
Method: In the current study, the apopto-
sis-related genes; FAS _670 A>G, FASLG
_844 C>T and FASLGIVS2nt _124 A>G
Poster Session 59 Autoimmunity: disease mechanisms
polymorphisms were genotyped in 120
Rheumatoid arthritis (RA) Iranian patients
and 112 unrelated healthy controls using
PCR-RFLP method.
Result: Our results demonstrated that
between the120 RA patients heterozygous
in FAS_670A/G (OR = 1.42, CI = 0.92
1.52, P = 0.18) and FASL_844C/T
(OR = 1.42, CI = 0.921.52, P = 0.18)
promoter region and homozygous in the
minor allele for FasLIVS2nt_124G/
G(OR = 1.43, CI = 0.761.81, P = 0.7)
polymorphisms were not associated with
elevated risk of RA in Iranian patient com-
pared to normal controls and although the
frequency of this polymorphisms is higher
in the cases than controls it is not statisti-
cally signicant.
Conclusion: We didnt found a statistically
signicant association between the Fas and
FasL polymorphisms and risk estimate of
RA in Iranian patient. To the best of our
knowledge, further investigation of these
polymorphisms with more cases is recom-
mended.
1422
Regulatory T cells in patients with
psoriasis and rheumatoid arthritis treated
by biological therapy
Kral, V1; Pohorska, J1; Filipovska, O2; Blazickova, S3;
Stiborova, I1; Burianova, Z2; Richter, J1
1
Institute of Public Health, Centre of Immunology and
Microbiology, Usti nad Labem, Czech Republic;
2
Masaryks hospital, Usti nad Labem, Czech Republic;
3
Trnavian University, Faculty of Health Care and Social
Work, Trnava, Slovakia
Background: Many studies have reported
numeric or functional regulatory T cells
(Treg) deciencies in various human auto-
immune diseases (AID). Both Treg and T-
helper 17 cells (Th17) play important roles
in the pathogenesis of psoriasis (PS). How-
ever, the relationship between Th17 and
Treg cells and their dynamic variations in
psoriasis remain unclear. Potential targets
in the treatment of this disorder include
biologic agents aimed at blockade of cyto-
kines, chemokines, and angiogenic factors.
Therapies that could possibly increase Treg
suppressive ability are proven to improve
the course of psoriasis and other AID. We
decided to determine and monitored Treg
in patients with Ps and rheumatoid arthri-
tis (RA) and tried to demonstrate inuence
of biological therapy (BT) on Treg and
Th17 cells proportion.
Method: Treg were determined using a
ow cytometry by characteristic marker of
Treg - CD127 and FoxP3 (CD127)CD25+
FoxP3+ of CD4+ lymphocytes) in patients
with Ps and RA treated with conventional
therapy (CT) (PSD, resp.RAD) and in
patients with BT (PSB, resp. RAB).
Allergy 66, Suppl. 94 (2011): 482642  2011 John Wiley & Sons A/S
Patients with PS were treated antiTNF
therapy similar to patients with RA. As
control group we used healthy volunteers
(HC). In patients with PS we determined
whether the number of Th17 dened as
IL17+ IFNgamma+ in CD4+CD3+ cells
and the effect of therapy was followed by
PASI scores.
Result: We observed the lowest proportion
of Treg in patients treated with CT - PSD
(5.5 2.3%), RAD (5.7 1.9%) and HC
(5.8 1.2%). Biological therapy signi-
cantly (P < 0.01) increased the number of
T reg in both groups. The highest number of
Treg was detected in the PSB (9.2 3.9%),
also in the RAB (8.0 3.3%). The lowest
value of PASI score was recorded in patients
treated with BT therapy (4.2 6.6)
compared with other groups of patients
with psoriasis (PSD group (10.5 7.9;
P < 0.05) and PSN (non-treated patients)
(14.1 9.1; P <0.01)). These ndings have
correlated with expected benecial effect of
biological therapy. Th17 in psoriasis showed
higher proportion in PSB (3.6 1.9%) and
PSD (3.2 2.7%) compared to HC (1.7
0.8%).
Conclusion: Patients with RA and psoriasis
on biological therapy have shown even sig-
nicantly higher Treg levels in comparison
to patients treated by conventional ther-
apy. There was shown decline of PASI and
Treg increase in psoriasis patients on bio-
logical therapy. The suggestion is, that bio-
logical therapy could positively inuence
the Treg proportion.
1423
Immunologic criteria of effective inflix-
imab therapy in RF-positive patients with
rheumatoid arthritis
Kalashnikova, T; Sizyakina, L
Rostov State Medical University, Department of Clinical
Immunology and Allergology, Rostov-on-Don, Russian
Federation
Background: The background of the study
was to research differences in cytokine pro-
le of responding and non-responding
patients with rheumatoid arthritis (RA) to
iniximab for development immunologic
criteria of effective anti-TNF-a therapy.
The method used studying of 33 RF-posi-
tive patients with RA, polyarthritis who
received therapy with methotrexate (12.5
15 mg) and Iniximab (3 mg/kg, nine infu-
sions). Examination of patients included
determination of IL-6, IL-1b, TNF-a,
RAIL-1 in serum of blood and spontane-
ous and PHA-stimulated supernatants of
PMBC by ELISA.
Results: The results obtained that in
patients with positive effect of chimeric
anti-TNF-a monoclonal antibody treat-
ment was estimated positive dynamic in
cytokine prole. Decreasing level of inam-
matory cytokines: IL-6 (initial level in
serum 17 5.6 pg/ml, in spontaneous su-
pernatants 5699.8 157 pg/ml, in stimu-
lated supernatants 9362.3 192 pg/ml;
after VI infusion level in serum
4.5 2.3 pg/ml, in spontaneous superna-
tants 533 21 pg/ml, in stimulated super-
natants 738 18 pg/ml), TNF-a (initial
level in serum 2.5 1.3 pg/ml, in sponta-
neous supernatants 80.8 13.7 pg/ml, in
stimulated supernatants 138.8 71.5 pg/
ml; after VI infusion level in serum
1.6 1.4 pg/ml, spontaneous supernatants
19.3 8.2 pg/ml, stimulated supernatants
22.3 9.1 pg/ml) and IL-1b (initial level
in serum 1.2 0.5 pg/ml, in spontaneous
supernatants 866.8 74.6 pg/ml, in stimu-
lated supernatants 1148.7 162.8 pg/ml;
after VI infusion level in serum
0.3 0.1 pg/ml, in spontaneous superna-
tants 316 35 pg/ml, in stimulated super-
natants 520 28 pg/ml) in combination
with good clinical response after VI infu-
sion could be used as accompaniments
immunologic criteria of effective Iniximab
therapy and possibility for cancellation
anti-cytokine therapy and use monothera-
py with methotrexate in these patients.
1424
The osteoprotegerin in synovial fluid
marker of the osteosynthesis in psoriatic
versus rheumatoid arthritis
Ciacli, C1; Gurban, C2; Puschita, M1; Cotoraci, C3;
Sfrijan, F2
1
Vasile Goldis West University, Department of
Immunology, Arad, Romania; 2Victor Babes University
of Medicine and Pharmacy, Department of
Biochemistry, Timisoara, Romania; 3Vasile Goldis West
University, Department of Hematology, Arad, Romania
Background: The aim of this study was to
investigate the osteoprotegerin levels in
synovial uid in psoriatic versus rheuma-
toid arthritis as a marker of osteosynthesis
at this level.
Method: Twenty-seven patients with psori-
atic arthritis and twenty-one patients with
rheumatoid arthritis were included. The os-
teoprotegerin levels in synovial uid were
quantied by using ELISA method (Endo-
gen, Woburg, MA, USA).
Result: The results of our study demon-
strates that osteoprotegerin concentration
in the synovial uid is higher in rheuma-
toid versus psoriatic arthritis. This corre-
sponds to the fact that bone destruction is
higher in rheumatoid arthritis, so that bone
formation is trying to counteract the effects
of bone damage.
Conclusion: Osteoprotegerin is an impor-
tant marker on the process of osteosynthe-
sis in psoriatic and rheumatoid arthritis
and prognostic value in detecting patients
539

who may benet from early specic ther-
apy before the onset of complications.
1425
Relationship between proinflammatory
cytokines and dual-energy X-ray absorp-
tiometry in psoriatic arthritis
Ciacli, C
Vasile Goldis West University, Department of
Immunology, Arad, Romania
Background: The aim of this study was to
investigate the serum levels of proinam-
matory cytokines (IL-1, IL-6, TNF-a) with
bone mineral density values in psoriatic
arthritis to identify useful correlations in
early diagnosis of complications and iden-
tify patients who may benet from specic
treatment.
Method: Twenty-seven patients with psori-
atic arthritis, twenty-one patients with
rheumatoid arthritis and twenty normals
were included. The pro-inammatory cyto-
kines levels were quantied by using
ELISA method (Endogen, Woburg, MA,
USA) and bone mineral density was mea-
sured at the lumbar spine (L2-L4) with
dual-energy X-ray absorptiometry
(DEXA).
Result: Our study results demonstrates that
there is a strong negative correlation
between blood values of proinammatory
cytokines and T score values in patients
with psoriatic and rheumatoid arthritis.
Conclusion: Our study justies the use of
DEXA in detecting and monitoring the
occurrence of osteoporosis in patients with
psoriatic and rheumatoid arthritis and
allows therapeutic intervention in patients
still asymptomatic preventing fractures,
redoubtable complications of the disease.
1426
Impact of anti-TNF-a monoclonal anti-
bodies on serum levels of cytokines
which regulate inflammation
Oransky, S1; Eliseeva, L1; Vasinova, J1; Khanferyan, R1;
DuBuske, L2
1
Kuban State Medical University, Krasnodar, Russian
Federation; 2Immunology Research Institute of New
England, Gardner, MA, United States
Background: Iniximab is a monoclonal
antibody that binds to TNF-a thereby
blocking receptor binding and clinical
effects of this cytokine. This study assesses
production of pro-inammatory TNF-a
and anti-inammatory IL-10 cytokines in
rheumatoid arthritis (RA) patients receiv-
ing inixamab.
Method: Twenty-ve patients with active
RA (average age 48.4 12.5 years; 19
female, 6 male) and a control group of 12
healthy volunteers were studied including
540
Poster Session 59 Autoimmunity: disease mechanisms
assessment of serum histamine, TNF-a and
IL-10 levels by ELISA. RA patients were
investigated before treatment and after ve
intravenous infusions of iniximab.
Result: In RA patients, the serum levels of
TNF-a and IL-10 before treatment were
much greater than in healthy controls with
TNF-a 178.1 (range 101243) and IL-10
302.5 (range 208.3510) pg/ml. Iniximab
signicantly decreased the concentration of
TNF-a by 2.4 fold (P = 0.02) but had no
effect on the serum concentration of IL-10.
Conclusion: The ability of iniximab to
reduce pro-inammatory cytokine levels of
TNF-a, rather than an impact on the anti-
inammatory cytokine IL-10, may in part
account for the anti-inammatory effects
of anti-TNF-a monoclonal antibodies in
RA patients.
1427
A case of periodic fever accompanied by
aphthous stomatitis, pharyngitis and cer-
vical adenitis syndrome, in an adult
Aguilera, R; Salinas, J
University of Chile, Clinical Hospital, Immunology
Section, Santiago, Chile
Background: The PFAPA (periodic fever,
apthous stomatitis, pharyngitis and adeni-
tis) syndrome is a chronic sporadic disease
characterized by abrupt onset of high fever
associate with aphthous stomatitis, tonsili-
tis, pharyngitis and cervical adenopathy.
This entity usually begins in childs under
ve years old and last between 3 and
6 days every 38 weeks. Patients are
asymptomatic between episodic intervals.
Currently, only few cases of this syndrome
in adults have been described.
Method: Here we report a case of a patient
with an 8 months history of presumed
PFAPA in an eighteen years old young
adult male.
Result: The patient has a history character-
ized by 6 episodes of fever, pharyngitis and
cervical adenopathy, lasting 6 days, with-
out clinical response to neither antibiotics
nor non-steroidal anti-inammatory drugs.
The laboratory test includes serial leuko-
cytes, bacterial culture, rheumatic disease
screening and Epstein Barr virus serology,
all with normal results. Common syn-
dromes associated with adult periodic
fevers were excluded based on the compila-
tion of this patients symptoms and labora-
tory evaluation. In addition, the patient
successfully responded to the published
treatment for PFAPA, a single dose of
60 mg of prednisone. Based on these nd-
ings, PFAPA diagnosis was nally made.
Conclusion: Therefore we concluded that
PFAPA should be considered in the differ-
ential diagnosis of adult-onset periodic
Allergy 66, Suppl. 94 (2011): 482642  2011 John Wiley & Sons A/S
fevers, however it remain a diagnosis of
exclusion.
1428
Juvenile dermatomyositis in association
with degos disease: a case report
Herrera, A; Yamazaki, M; Hernandez, V; Rodriguez, A;
Saez, M
INP, Clinical Immunology and Allergy, Mexico City,
Mexico
Background: A 7-year-old girl presented
with 8 months of fever, proximal weakness,
palpebral ptosis, heliotrope, hypertrichosis,
asthenia, adynamia, 4 kg weight loss and
papular, erythematous, umbilicated skin
injuries that left a pearly white scar involv-
ing the face, anterior and lateral thorax,
and lower extremities (thighs and knees). A
skin biopsy was performed reporting Der-
matopanniculitis interface vasculitis and
mucinosis consistent with Degos disease.
Method: Case report
Result: She presented with difculty swal-
lowing and reux and Gastrostomy with
Nissen was performed. Laboratory tests:
CK193, LDH 506, ANA++, IgG 2740,
EBV early antigen positive. Neuroconduc-
tion study: axonal motor polineuropathy.
Electromyography: myopathic pattern
compatible with myositis. An MRI: gener-
alized muscular atrophy. Muscular biopsy:
perivascular and perifascicular atrophy,
mild perimysial brosis and fat inltration,
regeneration and a perivascular lympho-
cytic inltrate between muscular fascicles.
Immunohistochemistry: LMP-1 lympho-
cytes in the interstice, 40% CD4 lympho-
cytes compatible with Dermatomyositis.
She received initially steroid at 0.5 mgkgd,
cyclosporine at 3 mgkgd, and IVGG infu-
sion at 0.7 grkgdo, with improvement in
strength, ptosis and resolution of skin inju-
ries, then she received anticoagulation ther-
apy (coumadin) and SC methotrexate at
0.2 mgkgsem. Cyclosporine, methotrexate
and anticoagulation were suspended
2 months later and mycophenolate mofetil
at 16 mgkgd was started, steroid dose was
increased to 0.8 mgkgd. She had a good
evolution, 2 months later after she was dis-
charged she was able to walk without any
help, ptosis improved, she had no skin
injuries but persists with hypertrichosis.
Conclusion: Degos disease is potentially
fatal, with a dark prognosis in most cases,
so the diagnosis and the opportunal treat-
ment becomes a challenge. It is a vasocclu-
sive disease, involving small and medium
sized arteries, affecting the skin, the gastro-
intestinal tract and the central nervous sys-
tem mainly. It is extremely rare, about 200
cases reported, 4 of them children. The age
at presentation is 35 years of age, more
frequent in males (1.33:1). In the last three
Poster Session 59 Autoimmunity: disease mechanisms

decades, at least six cases has been reported
with Degos disease in association with colla-
gen diseases, all females. It has been pro-
posed an autoimmune origin for these two
conditions, so the early and aggressive treat-
ment is essential for the outcome, as in our
patient, who had a good evolution.
1429
parameters, without typical clinical mani-
festations, should be evaluated for other
diseases primarily hematological malignan-
cies.
1430
Imbalance between Th17 and regulatory
T-cells in systemic lupus erythematosus
patients
inammatory response, and subsequent ex-
acerbatrion of the disease.
1431
Anti nuclear antibody test significance in
childhood
Ragusa, G; Rugiano, A; Celani, C; Caggiano, S; De
Castro, G; Indinnimeo, L; Duse, M
Sapienza University Hospital, Pediatrics, Rome, Italy

Non-Hodgkin lymphoma with laboratory
immunological analysis specific for Sjo-
gren syndrome a case report
Durutovic, D; Raskovic, S; Peric-Popadic, A; Stefanovic,
L; Tomic-Spiric, V; Djuric, V; Plavsic, A; Bogic, M
Clinical Center of Serbia, Clinic of allergology and
immunology, Belgrade, Serbia
Background: It is known that patients with
some autoimmune diseases, including Sjo-
gren syndrome have a higher risk for
development of lymphoproliferative dis-
eases.
Case report: A 66-year-old man was ad-
mited in our hospital to be evaluated for
Sjogren syndrome. He presented with
fever, fatigue, xerostomia, loss of appetite,
weight loss and severe dysphagia. Labora-
tory analysis showed high erythrocyte sedi-
mentation rate and high brinogen,
microcytic hypochromic anaemia, leucocy-
tosis, elevated levels of blood urea and cre-
atinine, elevated alkaline phosphates and
gamma-glutamyltransferase. Immunologi-
cal testing showed positive antinuclear
antibodies detected by indirect immunou-
orescence with epithelial cell line from
human laryngeal carcinoma as substrate
(IIF-HEp2) of 1:40 titer and positive anti-
Ro/SSA antibodies. Autoimmunity liver
disease parameters were positive: anti-
smooth muscle antibody (ASMA) of 1:320
titer and anti-mitochondrial (AMA) M2
antibodies in 1:40 titer. Immunoelectropho-
resis of serum proteins showed monoclonal
gammapathy (IgM kappa). Immunox-
ation of urine proteins detected monoclo-
nal kappa light chains and Bence Jones
proteins. Due to clinical presentation, eso-
phagogastroduodenoscopy was preformed
immediately after admission. It reveled ul-
cerostenotic tumor mass in the oesophag-
ogastric junction which spreaded in cardiac
region and fornix. Computer tomography
of abdomen showed the presence of tumor
mass in gaster, enlarged lymph nodes in
gastro-hepatic ligament and retroperitone-
aly. Histological and immunohistochemical
analysis of gastric tumor mass revealed the
presence of diffuse large B-cell non-Hodg-
kin lymphoma.
Conclusion: Presence of different autoanti-
bodies in patients with non-Hodgkin lym-
phoma is most probably due to polyclonal
stimulation of B lymphocyte. Patients who
have positive immunological laboratory
Allergy 66, Suppl. 94 (2011): 482642  2011 John Wiley & Sons A/S
Jakiela, B1; Plutecka, H2; Milewski, M2; Sanak, M2;
Musial, J2; Kleczynska, W1
Background: Anti nuclear antibody (ANA)
1
Department of Medicine, Jagiellonian University is a common test with uncertain clinical
Medical College, Allergy and Immunology Department, signicance. In order to evaluate the rele-
Krakow, Poland; 2Department of Medicine, Jagiellonian
University Medical College, Krakow, Poland
vance for differential diagnosis of rheu-
matic disease we collect results of ANA
and correlate it with nal diagnosis in 101
Background: An impaired function of reg-
ulatory T-cells (Treg) Leeds to a failure in children referred to our department for
suspected immunologic disease.
immune tolerance and triggers autoimmune
Method: A total of 101 children, aged
reactions. We studied whether in active
between 1 and 17 years (mean 7.1; median
systemic lupus erythematosus (SLE) a de-
5.8), 54 (53.5%) were boys, were studied
ciency in Treg takes place, and if it is
from 2008 to 2010. All patients underwent
accompanied by an increase in effector T-
ANA test for suspected transient hypo-
cell responses.
gammaglobulinemia (THI) and/or periodic
Method: We studied peripheral blood lym-
fever (53), rheumatic disease (27), IgA def-
phocytes (PBL) from seven patients with
iency (13) and urticaria (8). observed at
active SLE, eight patients with SLE in
1:80 dilution. For the statistical analysis,
remission, and eleven healthy controls. The
we used SPSS for Windows (version 9.0,
frequencies of effector IL-17A (Th17) and
SPSS, Chicago, IL, USA). Two by two
IFNgamma (Th1) producing CD4+ T-
tables were used to calculate correlation of
cells were assessed by ow cytometry after
the variables.
in vitro stimulation of PBL with phorbol
Result: A total of 72 patients (71.3%) were
myristate acetate and ionomycin. Circulat-
negatives, 23 (22.8%) were ANA positive
ing regulatory T-cells were evaluated as
with homogeneous pattern, 4 (4%) speck-
CD4+CD25high lymphocytes expressing
led, 2 (2%) nucleolar. 11% of patients with
FoxP3 antigen.
THI and/or periodic fever, 38% of patients
Result: There was no difference in the per-
with IgA deciency and urticaria were pos-
centage of CD4+CD25highFoxP3+ cells
itives. Only among patients resulted with
between the groups studied, however their
rheumatic disease a large proportion pre-
absolute counts were decreased in active
sented ANA positive test.
SLE (5.5 [17] cells/microliter) as com-
Conclusion: Final diagnosis of our study
pared to patients in disease remission (11
conrms that IgA deciency is often asso-
[615], P = 0.05) and healthy subjects (16
ciated with presence of innocent autoanti-
[10-20), P < 0.01). Both frequency and the
bodies: no IgA decient patient suffers
numbers of Th1 cells were signicantly
from autoimmune disease. Furthermore
decreased in patients with SLE (P < 0.01)
our data conrm that ANA test is not cru-
as compared to healthy controls, but no
cial even if it is more frequently associated
such difference was observed regarding the
with rheumatic disease. Only high titers
number of Th17 lymphocytes. In conse-
and homogeneous pattern of ANA are rel-
quence a decrease in Th1/Th17 ratio has
evant as marker of rheumatic disease.
been shown. A ratio of Treg to Th17 cells
in healthy controls (2.2 [1.83.6]) was
higher than in active SLE (1.1 [1.02.1],
P < 0.05), but not in inactive SLE (1.5 1432
[1.12.9]). Additionally, we observed a Prognostic value of anti-CCP and
clear negative correlation between the anti-MCV in undifferentiated connective
number of Tregs and markers of disease tissue disease
activity (e.g. SLEDAI score, r = )0.59,
Bodolay, E1; Csipo, I1; Tumpek, J1; Hajas, A1; Sipka, S1;
P < 0.01). Zeher, M1; Szegedi, G1; Szodoray, P2
Conclusion: Patients with active SLE are 1
University of Debrecen, 3rd Department of Internal
Medicine, Debrecen, Hungary; 2University of Oslo,
characterized by a deciency in Treg and
Institute of Immunology, Oslo, Norway
Th1 lymphocytes, which were both found
to inhibit Th17 response. These data sug- Background: To determinate the panel of
gest, that such imbalance between major autoantibodies of patients with undifferen-
T-cells subsets could lead to increased pro- tiated connective tissue disease (UCTD)
541

followed for at least 1 year. The sensitivity
and specicity of anti-MCV in comparison
with anti-CCP antibodies and RF were
determined in 243 patients with undifferen-
tiated connective tissue disease (UCTD).
Method: A total of 243 UCTD patients
with manifestations suggestive but not
diagnostic of specic connective tissue dis-
ease (CTD) were recruited and followed up
between 2008 and 2010. The patients sera
were tested for anti-nuclear (ANA), as well
as for ten different specic autoantibodies
(anti-dsDNA, -RNP, -SSA, -SSB, -Scl70, -
centromer, -cardiolipin [CL], -rheumatoid
factor, cyclic citrullinated peptide antibod-
ies (anti-CCP), and anti-modied citrulli-
nated vimentin (MCV).
Results: A total of 116 of the 243 UCTD
patients developed a dened CTD (42
rheumatoid arthritis [RA], 27 Sjogrens
syndrome, 10 systemic sclerosis, 23 mixed
connective tissue disease, 9 systemic lupus
erythematosus, and 5 antiphospholipid
syndrome). The presence of the anti-SSA
(OD: 2.279), anti-CL (OD:2.347), anti-
RNP (OD:2.208), anti-CCP (OD:16.304),
and anti-MCV (7.772) predisposed to tran-
sition into denitive CTD. In patients with
UCTD who progressed into RA, the sensi-
tivity of anti-MCV antibodies was 80.8%
with 90.9% specicity. For anti-CCP anti-
bodies the sensitivity was 47.0% and speci-
city 96.5%, while the sensitivity of RF
was only 33.3% with 42.2% specicity.
Anti-CCP and anti-MCV showed the high-
est sensitivity (81.8%), with 97.0% specic-
ity.
Conclusion: Anti-MCV was the most valu-
able marker in the early diagnosis of RA
in the UCTD stage. Combined detection of
anti CCP and anti MCTV increased the
542
Poster Session 59 Autoimmunity: disease mechanisms
diagnostic capability for progression of
RA.
1433
Polyostotic fibrous dysplasia. A difficult
differential diagnosis in patient with au-
toantibodies
Caggiano, S1; Celli, M2; Leonardi, L2; Roggini, M2;
Ragusa, G3; Zicari, A3; Duse, M3
1
Umberto I Hospital Sapienza Rome University,
Pediatric Immunology and Allergology, Roma, Italy;
2
Umberto I Hospital Sapienza Rome University,
Pediatiric, Rome, Italy; 3Umberto I Hospital Sapienza
Rome University, Pediatiric Immunology and
Allergology, Rome, Italy
Polyostotic Fibrous Dysplasia of bone is a
rare non-inheritable congenital disease in
which bone is replaced by dysplastic
brous tissue. It is classied in accord to
the number of affected bones and its asso-
ciation to endocrine defect as monostotic,
polyostotic and Albrights disease. We
report the case of A 13-year-old male
patient diagnosed as LES because of his
antinuclear antibodies positivity. The boy
reported recurrent articular pain, not asso-
ciated to signs of objective arthritis neither
increased inammatory markers. Family
history was signicant for autoimmune dis-
eases: maternal grandmother and aunt
reported chronic lymphocytic thyroiditis;
mother reported recurrent abortions due to
natural killer cells hyperactivity without
autoimmune markers. The child has been
healthy until the age of 11 when he
reported two episodes of left knee pain and
fever. 3 months later the patient presented
left sternoclavicular joint swelling with
fever and 8 months later he had pain at
the left proximal humeral diaphysis, site of
Allergy 66, Suppl. 94 (2011): 482642  2011 John Wiley & Sons A/S
previous fracture. On clinical examination
were found a 40 mm cafe-au-lait hyperpig-
mentation of the necks skin, permanent
exuberant pathological callus of the proxi-
mal diaphysis of the left humerus with
hypomobility and pain at the active and
passive mobilization. Laboratory tests
showed elevated inammatory markers.
Bone scan described hyperxation sites in
the cranial portion of the left humeral
diaphysis, site of previous fracture and in
anterior arch of the fourth right rib. A
scheletal X-ray showed osteolytic area with
periosteal reaction on the anterior arch of
IV right and in the in the callus of left
humerus. Bone marrow aspiration excluded
multiple myeloma, Langerhans cell histio-
cytosis or other lymphoproliferative dis-
ease. MRI showed pathological tissue in
the fracture site extended to periosteum.
Bone biopsy in this area excludeds cancer
but showed remodelling and focal bone
apposition, compatible with a polyostotic
brous dysplasia. Biochemical investigation
showed high serum osteocalcin and calciu-
ria. Densitometry highlighted osteopenia
with a T score of )1.4. The diagnosis of
polyostotic brous dysplasia was conrmed
and therapy with bisphosphonates and cal-
cium was promptly started and the inam-
matory markers have been progressively
normalized. This case emphasizes the risk
of giving diagnostic value to laboratory
exams as autoantibodies when it is known
that they can occur as innocent ndings
without any biological signicance.

Immunodeficiency: the role of T cells
1434
Unusual association between three
different genetic diseases
Ragusa, G; Iacobini, M; Antonetti, L; Celani, C; Barbato,
M; Caggiano, S; Duse, M
Sapienza University Hospital, Pediatrics, Rome, Italy
We reported the case of a 5 year child
affected by Chronic Granulomatous Dis-
ease (CGD) where the diagnosis has
been delayed by the previous onset of
Holt-Oram Syndrome associated to laryn-
gomalacia. At born he was diagnosed as
Holt-Oram Syndrome because of presence
of transitory ventricular septal defect, bid
thumb, index and little ngers agenesis in
right hand, index nger agenesis in left
hand. Familiar history revealed two mis-
carriages (grandmother and mother) and
Hashimoto Thyroiditis (mother). Diffuse
pyoderma was reported before the dis-
charge. resistant sideropenic anemia until
2 years when duodenal biopsy conrmed
Celiac disease showing MARSH type I;
also HLA DQ2 was positive. in this occa-
sion, a routine chest X ray revealed a
severe unilateral pneumonia with pleura
effusions, and very high inammatory
markers. A relapse of pneumonia was
reported in the next year. When 5, he pre-
sented gastroenteritis associated to ulcer-
ated nodules on the suspected as cutaneous
metastasis of Chrons disease. Colonoscopy
showed normal mucosa with many intrapa-
rietal granulomas. At the our rst exami-
nation the child was in good general
condition but Blood tests showed neutro-
philia, thrombocytosis, iron deciency,
microcytic hypochromic anemia and persis-
tence of high inammatory markers. We
suspected Chronic Granulomatous disease
(CGD) and performed Nitroblue tetrazo-
lium (NBT) test that resulted negative,
conrming the diagnosis. Mothers NBT
showed the reduction of NBT in 50%
PMN. Clinical picture and laboratory data
were compatible with CGD which is
marked by failure to destroy bacteria and
fungi phagocytosed by neutrophils and
macrophages. The immunodeciency is
characterized by an abnormal NADPH
oxidase, due to four different genetic
defects: deciency of the heavy chain in the
cytochrome (X-linked) is the most fre-
quent; molecular analysis is still ongoing.
The patients meanwhile began antibiotic
Allergy 66, Suppl. 94 (2011): 482642  2011 John Wiley & Sons A/S
Poster Session 60
prophylaxis with Cotrimoxazole 5 ml per
day. After only 3 weeks cutaneous nodules
disappeared and inammatory markers
constantly high in the last 3 year normal-
ized.
1435
Consanguineous marriages in families
with 1 or more than 1 child with autoso-
mal recessive chronic granulomatous dis-
ease
Modarres Saryazdi, Z1; Pourpak, Z1; Fazlollahi, M1;
Maddah, M1; Sedighipour, L1; Fattahi, F1; Nabavi, M2;
Bazargan, N3; Movahedi, M4; Moin, M1
1
Tehran University of Medical Sciences, Immunology,
Asthma and Allergy Research Institute, Tehran, Islamic
Republic of Iran; 2Semnan University of Medical
Sciences, Department of Pediatrics, Semnan, Islamic
Republic of Iran; 3Kerman University of Medical
Sciences, Department of Pediatrics, Faculty of
Medicine, Kerman, Islamic Republic of Iran; 4Tehran
University of Medical Sciences, Department of
Immunology and Allergy, Childrens Medical Center,
Tehran, Islamic Republic of Iran
Background: Chronic granulomatous dis-
ease (CGD) which caused by a defect in
NADPH oxidase enzyme in leukocytes is a
rare hereditary primary immunodeciency
(PID) disorder characterized by recurrent
bacterial and fungal infections. CGD may
be inherited in an X-linked or autosomal
recessive (AR) form. The aim of this study
was to investigate the prevalence of con-
sanguineous marriages in families with one
or more than one child with AR CGD.
Method: Ninety one families with 134 AR
CGD children (mean age 14.52 8.81
years and male/female ratio 1.17) were
included in this study. The diagnosis was
based on the clinical ndings of CGD and
results of NBT and DHR test. Subjects
were categorized in two groups of families
with at most one child of CGD (group I)
or more than one child with CGD (group
II). Each group was sub classied based on
the consanguinity (rst cousin and beyond
rst cousin) of the parents.
Result: Group I consisted of 66 patients
from 66 families and group II consisted of
68 patients from 25 families. The frequency
of the rst cousin, beyond rst cousin and
non consanguineous marriages in group I
was 55.6%, 22.2% and 22.2% respectively.
In group II, 40 patients (58.8%) from 16
families and 24 patients (35.3%) from 7
families were reported to have rst cousin
and beyond rst cousin consanguineous
parents while four patients (5.9%) from
two families were reported to have non
consanguineous parents. Of the 25 families
in group II, 21 families had consecutive
CGD siblings. The overall, rate of consan-
guineous marriages was 86.4%, which was
signicantly (p=0.007) higher in group II
(94.1%) compared to group I (77.8%).
First cousin marriages were more signi-
cantly common in parents of CGD patients
compared to beyond rst cousin and non
consanguineous marriages (P < 0.0001).
Conclusion: Our results showed the high
prevalence of consanguineous marriages in
families with CGD children especially
those with more than one child with CGD.
It is recommended to warn families with
consanguineous marriages and particularly
rst cousin marriages about the risks of
genetically inherited disease. Consanguine-
ous families with one CGD child should
also be aware of the high risk of CGD
occurrence in their following children.
1436
Comparative analysis of the HLA-B*5701
allele frequencies among HIV-infected
patients in Moscow and Ekaterinburg
areas, Russia
Lebedeva, N1; Serkov, I2; Pronin, A3; Sandyreva, T4;
Gudima, G5; Sidorovich, I6
1
Moscow regional Center of AIDS and Infectious
Diseases Prevention and Treatment, Clinical Dept.,
Moscow, Russian Federation; 2Moscow regional Center
of AIDS and Infectious Diseases Prevention and
Treatment, Diagnostics Dep., Moscow, Russian
Federation; 3Moscow regional Center of AIDS and
Infectious Diseases Prevention and Treatment, Head,
Moscow, Russian Federation; 4Sverdlovsk regional
Center of AIDS and Infectious Diseases Prevention and
Treatment, Clinical Diagnostics Dept., Ekaterinburg,
Russian Federation; 5Institute of Immunology, Lab. of
Physiology of Immunity and Allergy, Moscow, Russian
Federation; 6Institute of Immunology, Head of AIDS
Immunobiology Dept., Moscow, Russian Federation
Background: Abacavir is a nucleoside
reverse transcriptase inhibitor used for
combination antiretroviral therapy (ART)
for treating human immunodeciency virus
(HIV) infection. An adverse effect from ab-
acavir is a treatment-limiting hypersensitiv-
ity reaction (HSR), which can be severe
and potentially life-threatening. Abacavir-
induced hypersensitivity reaction has been
associated with the presence of the major
543

histocompatibility complex class I allele
HLA-B*5701. Estimation of HLA-B*5701
allele distribution among HIV-infected
patients is very important for effective
ART with decreased adverse effects caused
by individual sensitivity to abacavir.
Method: A total of 184 blood samples
were collected from HIV-infected patients
in Moscow area and 817 blood samples
were collected from HIV-infected patients
in Ekaterinburg city and Sverdlovsk area.
HLA-B*5701 allele detection in collected
samples was performed by polymerase
chain reaction (PCR) in real time mode. CD4 lymphocyte counts were 750
Result: In samples collected in Moscow
region 8 samples were HLA-B*5701-posi-
tive, 171 were HLA-B*5701-negative, 5
were undetermined. So the frequency of
HLA-B*5701 allele among HIV infected
patients in Moscow area is 4.46%. In
samples collected in Ekaterinburg and
Sverdlovsk area 47 samples were HLA-
B*5701-positive and the rest 447 samples
were negative. The frequency of HLA-
B*5701 allele presence among HIV infected
of Ekaterinburg city and Sverdlovsk area
is 5.75%. Statistically proved differences
between frequency of HLA-B*5701 in
patients in two investigated areas were not
found (P = 0.5, according to the chi-
square criteria).
Conclusion: Our results are completely
consistent with available data on the distri-
bution of HLA-B*5701 allele in a popula-
tion of Caucasians. The HLA-B*5701
screening test minimizes potential adverse
effects of abacavir by identifying patients
who may be at risk of developing a hyper-
sensitivity reaction.
1437
Changes of CD4 cells levels after high
active antiretroviral therapy in HIV
infected children in Moscow region
Smirnov, E1; Drobyshevskaya, E1; Pronin, A1; Kaminsky,
G1; Gudima, G2; Sidorovich, I3
1
Moscow regional Center of AIDS and Infectious
Diseases Prevention and Treatment, Clinical Dep.,
Moscow, Russian Federation; 2Institute of Immunology,
Lab. of Physiology of Immunity and Allergy, Moscow,
Russian Federation; 3Institute of Immunology, Dept. of
AIDS immunobiology, Moscow, Russian Federation
Background: A total of 284 HIV-infected
children (vertical transmitted infection)
born in 19982010 are registered in Mos-
cow region. 161 of them (57%) receive
continuous high active antiretroviral ther-
apy (HAART). Here we analyzed the
dynamics of CD4 lymphocytes in these
children during the course of HAART.
Method: CD4 lymphocyte count were per-
formed by ow cytometry. Primary medical
documentation search and HIV-infected
patients register database of Moscow
region AIDS Info System were used.
544
Poster Session 60 Immunodeciency: the role of T cells
Result: More than a half (59.2%) of HIV-
infected children are need in HAART dur-
ing the 1st three years of life. About 5% of
HIV-infected children before 9 years old
have stable clinical-laboratory parameters
and are not need in HAART. HAART
started at 1st year of life in 30.9% of HIV-
infected children, at 2nd year of life in
17.1%, at 3rd in 11.2%, at 4th in
13.2%, at 5th in 6.6%, at 6th in 9.2%,
at 7th in 4.6%, at 8th in 3%, at 9th
at 5.3% In children aged before 1 year
(normal CD4 count 15002800 cells/mcl)
nausea, vomiting followed by jaundice and
1490 cells/mcl in middle suppression,
<750 cells/mcl in severe suppression. In
children aged 15 years (normal CD4
count 10001800 cells/mcl) CD4 lympho-
cyte counts were 500990 cells/mcl) in mid-
dle suppression, <500 cells/mcl) in severe
suppression. In children elder 5 years (nor-
mal CD4 count 5001100 cells/mcl) CD4
lymphocyte counts were 200490 cells/mcl
in middle suppression, <200 cells/mcl in
severe suppression. In children with middle
immunosuppression the recovery of CD4
count was achieved after 3.3 months (1st
year of life), after 7.7 months (age 1
5 years), and after 8.5 months of HAART
(age >5 years). In children with severe
immunosuppression the recovery of CD4
count was achieved after 10.3 (age 1
5 years) and after 9.2 months of HAART
(age >5 years) respectively. In three cases
of severe immunosuppression there was no
complete recovery of immune status after
1416 months of HAART.
Conclusion: In HIV-infected chlidren with
middle immunosuppression the recovery of
CD4 lymphocyte counts was achieved fas-
ter (after 3.3 months of HAART) at 1st
year of life then in children elder 1 year
(after 78 months of HAART). The recov-
ery of CD4 lymphocyte counts in HIV-
infected children elder 1 year with severe
immunosuppression was achived after 9
10 month of VAART. 2% of HIV-infected
children with severe immunosuppression
needs the prolonged period of HAART.
1438
Completely impaired degranulation and
cytotoxicity in two siblings with Chediak-
Higashi syndrome
Badawy, S1; Badr, M1; Hassan, T1; Ehl, S2; Maul-Pavicic,
A2
1
Zagazig University, Department of Pediatrics, Zagazig,
Egypt; 2University of Freiburg, Centre of Chronic
Immunodeficiency, University Medical Center, Freiburg,
Germany
Background: Chediak-Higashi syndrome
(CHS) was described by Cesar in 1943,
Chediak in 1952, and Higashi in 1954.
Chediak-Higashi syndrome is a rare child-
Allergy 66, Suppl. 94 (2011): 482642  2011 John Wiley & Sons A/S
hood autosomal recessive disorder that
affects multiple systems of the body.
Patients with Chediak-Higashi syndrome
exhibit hypopigmentation of the skin, eyes,
and hair;recurrent infections;abnormal nat-
ural killer cell function;and peripheral neu-
ropathy.
Case-1-boy History:Male, A.R.M, 4 years,
1st kid, from Ismailia (Egypt), positive con-
sanguinity. Condition started with bloody
diarrhea at 1.5 years and during next
1.5 years, he had recurrent throat infection
with fever. At 3 years, the patient developed
abdominal enlargement then oliguria. Then
pt had severe recurrent abscesses with pain
and difculty during sitting and standing.
Examination: Generalized skin darken-
ing (mainly extremities) like-dirt, change
hair color specially forelock (black to gray
and white), two face abscesses (nose and
cheeks), horizontal nystagmus since birth,
hepatosplenomegaly and massive ascitis.
Case-2-girl (boys sister) History:Female,
A.R.M, 2 years, 2nd kid, from Ismailia
(Egypt), positive consanguinity. Condition
started at 3 months with constipation and
recurrent throat infections and fever
responding to antibiotics and antipyretics.
Examination: Darkening of the skin
(mainly in face) with change hair color spe-
cially forelock (black to silver-gray and
white).
Results (work up): CBC: cytoplasmic inclu-
sions in lymphocytes, monocytes and neu-
trophils.
Bone Marrow: Hypercellular BM with
vacuoles and giant granules in cytoplasm
of eosinophils, eosinophilic myelocytes and
lymphocytes.
HLA Matching Testing:
Boy: A (A29), B (B7, B62, BW6), C
(CW1, CW6).
Girl: A (A2, A34), B (B41, B50,
BW6), C (CW2, CW6).
Father: A (A29, A34), B (B41, B62,
BW6), C (CW1, CW2).
Mother: A (A2, A29), B (B7, B50,
BW6), C (CW6).
Degranulation assay:
NK CD107 Degranulation: absent
degranulation of NK cells after stimu-
lation with target cells. NK CD107
after IL-2 stimulation: strongly
impaired reconstitution of NK
degranulation after pre-stimulation
with IL-2. CTL CD 107: reduced
baseline expression of CD 107 on
CTL blasts, but normal relative
increase upon stimulation.
Cytotoxicity assay:
Consistent with absent NK degranula-
tion, absent cytotoxicity of NK cells
on target. Normal cytotoxicity of
CTL blasts after stimulation with
anti-CD3/CD28.
Poster Session 60 Immunodeciency: the role of T cells
Differential diagnosis: (i) Albinism (ii)
Bacterial mouth infections (iii) Cutaneous
T-cell lymphoma (iv) Griscelli syndrome
(v) Pyoderma gangrenosum
1439
Cytomegalovirus-specific T lymphocytes
and lymphocyte subpopulations in
patients after kidney transplantation
Hanzlikova, J1; Vlas, T1; Reischig, T2; Panzner, P1
1 patients with persistent leukocytosis in
Pilsen University Hospital, Immunology and
Allergology, Pilsen, Czech Republic; 2Pilsen University
Hospital, Internal Clinic, Pilsen, Czech Republic
Background: We investigated the incidence
of the CMV (cytomegalovirus) infection
and immune response to it in patients after
kidney transplantation. We determined the
relative number of T cells producing intra-
cellular IFN-c (interferon-gamma) after
stimulation with CMV antigen (% CMV-
specic T lymphocytes) and absolute num-
ber of CD4+, CD8+ and CD19+
(CD4+, CD8+, CD19+/ul) in CMV-
positive (CMV+) and CMV negative
(CMV-) patients before during and after
transplantation.
Method: 1) Intracellular production of
IFN-c by CMV-specic T cells was deter-
mined by ow cytometry after incubation
of cells with CMV antigen using monoclo-
nal antibodies a-IFN-c, a-CD45 and a-
CD4.
2) T lymphocytes CD4+, CD8+ and B
lymphocytes CD19+ were determined by
standard ow cytometry.
3) PCR method was used to detect
CMV positivity or negativity. Patients: 21
patients, 16 men and 5 women, mean age
46.9 years. Patients were examined before
and during 12 months after kidney trans-
plantation. Seven patients were found
CMV positive and 14 patients were found
CMV negative before transplantation.
Result: 1) Before transplantation: Mean
absolute number of CD19+ cells in
CMV+ patients was 183.3, in CMV- 68.5,
P = 0.0137.
2) Five months after transplantation: a)
Mean absolute number of CD4+ cells in
CMV+ patients was 291.8, in CMV-
1017.7, P = 0.0045. b) Mean absolute
number of CD8+ cells in CMV+ was
396.6, in CMV- 668.0 P = 0.0510.
3) Before and during12 months after
transplantation: a) Mean absolute number
of CMV-specic T cells was in CMV+
2.31 and in CMV- 1.49, P = 0.0300. b)
Mean absolute number of CD4+ was in
CMV+ 1198.6, in CVM- 1567.8,
P = 0.0002. Other measured values were
not statistically signicant.
Conclusion: Signicant differences of the
counts of T lymphocytes were observed in
CMV+ and CMV- patients after kidney
Allergy 66, Suppl. 94 (2011): 482642  2011 John Wiley & Sons A/S
transplantation, counts of both helper and
cytotoxic T lymphocytes being lower in
CMV- patients in comparison with
CMV+ patients. Counts of CMV-specic
T lymphocytes were higher in CMV+
patients compared to CMV- patients only
in measurement performed 12 months after
transplantation. Supported by VZ MSM
0021620812
1440
A rare immunodeficiency. Leukocyte
adhesion deficiency type-3
Cavkaytar, O1; Buyukavci, M2; Aytac Elmas, S3;
Gumruk, F3; Sanal, O1
1
Hacettepe University Ihsan Dogramaci Childrens
Hospital, Pediatric Immunology and Allergy Unit,
Ankara, Turkey; 2Ataturk University Hospital, Pediatric
Allergy Unit, Erzurum, Turkey; 3Hacettepe University
Ihsan Dogramaci Childrens Hospital, Pediatric
Hematology Unit, Ankara, Turkey
Background: Leukocyte adhesion de-
ciency-3 (LAD-3) is a rare immunode-
ciency characterized by leukocytosis,
platelet dysfunction, and recurrent infec-
tions, resulting from dysfunctional signal-
ing via leukocyte integrins. A mutation in
FERMT-3 gene encoding kindlin-3 protein
is known to be the etiology of the disease.
Clinically, LAD-3 patients present with
symptoms similar to a combination of
LAD-1 and Glanzmanns thrombasthenia
and LAD-3 is described to be a variant of
LAD-1. Since 1970s, over 200 cases of
LAD-1 have been described. LAD-3 has
been described in approximately 20
patients worldwide. Here is a new patient
described.
Case presentation: A two month old girl
referred to our hospital with a history of
recurrent petechiae and echymoses on her
extremities started soon after birth and a
history of hospitalization for 40 days due
to fever and persistant leukocytosis. Par-
ents were 2nd degree relatives and a male
sibling who had bleeding diathesis died at
35th day after birth. On physical examina-
tion the umbilical cord was still unde-
tached she had multiple petechiae and
ecchymosis on her extremities. Laboratory
examination revealed anemia (Hemoglobin:
7.7 g/dl), leukocytosis (white blood cell:
27000 /ll) which persisted during the fol-
low-up period and mild thrombocytopenia
(thrombocyte: 78000 /ll). CD 18 level on
leukocytes was normal. On peripheral
smear 52% of the cells were lymphocytes,
25% were neutrophils, 8% atypical imma-
ture cells, 3% were monocytes and the rest
were early myeloid cells. Bone marrow
aspiration was cellular and found to be
normal and there was no arrest in any cell
series. A clinical diagnosis of LAD-3 was
suspected and molecular analysis of FER-
MT3 gene revealed a homozygous muta-
tion in exon 12 (c.1525C>T). The parents
and the healthy looking sibling were found
to be carriers. She experienced recurrent
pulmonary infections during the following
three months. She is going to be given
bone marrow from her mother who is fully
matched for HLA A, B, C, DR and DQ
allells soon.
Conclusion: LAD-3 is one of the rarer
immune deciencies with recurrent infec-
tions starting from early infancy. In
between infections and bleeding diathesis
the diagnosis should be considered. Alloge-
neic bone marrow transplantation is the
only chance for denitive treatment and
should be considered as early as possible
before the recurrence of infections.
1441
Detection of markers of acute stage of
herpes virus infections in HIV-infected
patients
Trubcheninova, L1; Gudima, G2; Trefilieva, N1;
Ishchenko, M1; Nikolaeva, I3; Ilina, N4; Sidorovich, I5;
Khaitov, R6
1
Institute of Immunology, Lab. of Secondary
Immunodeficiencies, Moscow, Russian Federation;
2
Institute of Immunology, Lab. of Physiology of
Immunity and Allergy, Moscow, Russian Federation;
3
Institute of Immunology, Lab. of Biotechnology and
AIDS, Moscow, Russian Federation; 4Institute of
Immunology, Clinical Dep., Moscow, Russian
Federation; 5Institute of Immunology, AIDS
Immunobiology Dep., Moscow, Russian Federation;
6
Institute of Immunology, Head, Moscow, Russian
Federation
Background: Detection of acute stage of
herpes virus infection connected with IgM
and active viral replication (HSV 1 and 2,
CMV, EBV). HSV-specic IgM occured
only at acute period of disease during
about 2 weeks, so detection of virus-spe-
cic IgM is important and valuable for
diagnostics.
Method: Fifty sera samples from drug-
dependent people were investigated with
enzyme immunoassay: 12 - from patients
HIV-infected during 1-1.5 years (group 1),
8 - from patients HIV-infected >3 years
(group 2) and 30 - from HIV-uninfected
persons (group 3).
Result: HSV-specic IgM antibodies in
group 1 were detected in 16.6%, in group
2 - in 25%, in group 3 - in 43% of sam-
ples. CMV-specic IgM antibodies in
group 1 were detected in 8.3%, in group 2
- in 12.5%, in group 3 - in 6.6% of sam-
ples. EBV-specic IgM antibodies were not
detected in groups 1 and 2, and in 3.3% of
samples in group 3. So detection of HSV-
and CMV-specic IgM antibodies was the
most prominent in group of drug-depen-
dent patients with long perion of HIV
infection. There were not detected IgM
antibodies against capsid antigen (VCA) of
545

EBV. IgG antibodies specic to early anti-
gen (EA), which is a marker of reactiva-
tion, were detected in 50% of samples
from group 1, in 62.5% of samples in
group 2 and 16% of samples in group 3.
Also high titers of EBV nuclear antigen
(EBNA)-specic IgG antibodies were
detected at the same samples.
Conclusion: At background of immunode-
ciency caused by HIV infection there is
activation of herpes viruses which leads to
acute form of disease. Such patients are
need in additional investigation and anti-
HSV therapy. Study of the interaction of
HIV and other pathogenic microorganisms
is very important for the development of
new strategies of prophylactics and therapy
of HIV infection.
1442
A case of DOCK8 deficiency and Epstein-
Barr virus associated intracranial lym-
phoma
Kara Eroglu, F1; Cavkaytar, O2; Cagdas Ayvaz, D2; Karly
Oguz, K3; Uner, A4; Varan, A5; Tezcan, I2; Sanal, O2
1
Hacettepe University Ihsan Dogramaci Childrens
Hospital, Department of Pediatrics, Ankara, Turkey;
2
Hacettepe University Ihsan Dogramaci Childrens
Hospital, Department of Pediatric Immunology, Ankara,
Turkey; 3Hacettepe University Faculty of Medicine,
Department of Radiology, Ankara, Turkey; 4Hacettepe
University Faculty of Medicine, Department of
Pathology, Ankara, Turkey; 5Hacettepe University
Institude of Oncology, Department of Pediatric
Oncology, Ankara, Turkey
Background: Combined immunodeciency
with DOCK8 mutation is a newly dened
form of autosomal recessive Hyper IgE
syndrome. Up to now, nearly 30 patients
has been reported in literature who have
susceptibility to recurrent sinopulmonary
and viral infections. Susceptibility to devel-
opment of cancers has also been dened in
DOCK8 deciency and different kind of
cancers such as cutenous T cell lymphoma-
leukemia, Burkitt lymphoma and meta-
static squamous cell carcinoma have been
reported. Herein we report an eleven years
old girl with DOCK8 deciency who has
been diagnosed as Epstein-Barr virus
(EBV) associated lymphoma after a long
follow up with the diagnosis of brain
abscess.
Case: An eleven year old girl admitted to
our clinic with complaints of recurrent
fever, cough and granular necrotic lesions
on her left buccal mucosa and gingiva.
Physical examination was unremarkable
except cutenous lesions on her chest, con-
sistent with epidermodysplasia verrucifor-
mis, buccal lesion and spastic ataxia.
Laboratory examinations revealed eosino-
philia, low CD4 (300 /mm3) count, high Ig
A (438 mg/dl), high Ig E (>5000 mg/dl)
and low IgM levels (22 mg/dl). Mutation
analysis revealed DOCK8 deciency. Five
546
Poster Session 60 Immunodeciency: the role of T cells
months after diagnosis, she admitted to
our emergency clinic with complaints of
fever and impaired consciousness. Brain
MRI revealed solitary mass with peripheral
edema consistent with brain abscess on left
parietal lobe. She has been treated with
wide spectrum antibiotics, antifungals and
steroids for ve months. Despite these
treatments, her lesion did not decrease in
size and histopathological examination of
the sample obtained by craniotomy was
consistent with diffuse large B cell lym-
phoma. Tumor cells were positive for EBV
EBNA antigen. Computed tomography of
abdomen and thorax did not show any
metastasis. She was diagnosed grade IV
lymphoma. She has been on chemotherapy
and radiotherapy for the last four months
without any neurological improvement.
Conclusion: To our knowledge this is the
rst case of EBV associated intracranial
lymphoma dened in a patient with
DOCK8 deciency.
1443
Secondary combined immunodeficiency
in patients with hypoplastic left heart
syndrome 2 case reports
Hlavackova, E1; Jienska, H2; Humlova, D2; Navratil, J2;
Vlkova, M3
1 indicated. Each infection episode must be
St. Annes Faculty Hospital, Department of Clinical
Immunology and Allergology, Brno, Czech Republic;
2
Brno Faculty Hospital, Pediatric Department
Department of Pediatric Cardiology, Brno, Czech
Republic; 3St. Annes Faculty Hospital Brno,
Department of Clinical Immunology and Allergology,
Brno, Czech Republic
Background: Secondary combined immu-
nodeciency due to protein-losing enterop-
athy is a common but underestimated
complication in patients suffering from
hypoplastic left heart syndrome. This syn-
drome results from rare cyanotic inborn
heart anomalies with incidence of 12% of
all inborn heart diseases. In consequence it
leads to diastolic dysfunction with labora-
tory results similar to those in primary
intestinal lymfangiectasia.
Method: We present case reports of two
patients suffering from protein-losing
enteropathy due to a chronic congestive
heart failure which developed after series
of palliative reconstructive heart surgeries
for hypoplastic left heart syndrome. Both
patients were referred because of frequent
respiratory tract infections, but not oppor-
tunistic infections, aged 5 and 17 years.
Result: Immunological investigation revealed
severe hypogammaglobulinaemia. In the
younger patient initial IgG level was 1.8 g/l
(ref. range 7.0014.00 g/l), while his IgA
and IgM levels were within reference ranges.
During his 4 years follow-up IgG decreased
to 0.73 g/l, such as IgA 0.44 g/l (ref. range
0.702.30 g/l) and IgM 0.37 g/l (ref. range
Allergy 66, Suppl. 94 (2011): 482642  2011 John Wiley & Sons A/S
0.401.5 g/l). Immunological investigation
of the second patient revealed decreased
IgG (2.76 g/l) and IgA (0.38 g/l) levels,
while his IgM was within the reference
range. Both patients had marked lymphope-
nia 0.3 g/l and 0.23 g/l respectively. Lym-
phocyte subsets determination revealed
markedly reduced relative numbers of
CD4+ cells. CD8+ cells remained within
normal range in relative numbers, CD19+
and NK cells were increased. In both
patients a marked decrease of the absolute
number of the cells was present. One patient
showed a good response to vaccination by
protein antigen (tetanus toxoid). Vaccina-
tion with pneumoccocal polysaccharide
antigen resulted in increase of specic IgG
levels but did not reach the reference range
and subsequently decreased in two years fol-
low-up. The initiations of ko-trimoxazole or
amoxycilin-clavulanic acid prophylaxis lead
to reduction of the number of respiratory
tract infections. The younger patient is still
alive. The older patient died aged 18 of
Campylobacter enteritidis complicated by
cardiac failure.
Conclusion: Patients with symptoms of
protein-losing eneteropathy of any origin
should be referred to immunologist in
order to monitor their immunological sta-
tus and start antibiotic prophylaxis when
aggressively treated with antibiotics.
1444
Two siblings with familial hemophagocy-
tic lymphohistiocytosis: Compound het-
erozygosity with two novel PRF1
mutations
Metin, A1; Akan, A1; de Saint basile, G2; Can Naci, K1
1
Ankara Hematology Oncology Childrens Research and
Taining Hospital, Pediatric Allergy and Immunology,
Ankara, Turkey; 2Hopital Necker-Enfants Malades, Lab.
dImmuno-Hematologie Pediatrique, Paris, France
Background: Familial hemophagocytic
lymphohistiocytosis (FHL) is a rare disease
with autosomal recessive inheritance.The
clinical and laboratory ndings may not
help to distinguish primary (FHL) and sec-
ondary hemophagocytic lymphohistiocyto-
sis(HLH). In this study, two siblings of
unrelated parents with the same, novel
compound heterozygous missense muta-
tions of perforin (PRF1), but with different
age of onset are reported.
Patient 1 (P1): An 8 year old girl was
referred with prolonged high fever. She
was the rst child of the parents. Because
of hepatosplenomegaly, pancytopenia, hy-
perferritinemia, hypertriglyceridemia and
hypobrinogenemia, bone marrow aspira-
tion was performed and she was diagnosed
HLH. With the high level of IgM for viral
capcid antigen (VCA) of EBV, it was
Poster Session 60 Immunodeciency: the role of T cells
thought to be secondary to EBV infection,
also because of the absence of any known
PRF1 mutations. She died of multiple
organ failure despite chemotherapy.
Patient 2 (P2): Five years later the death
of P1, the 7 month old son of the family
admitted with the same complaint, similar
clinical and laboratory ndings with hemo-
phagocytosis on bone marrow sample. All
viral tests were negative. During the ther-
apy, HLH was reactivated. The genetic
search for Familial Hemophagocytic lym-
phohistiocytosis (FHL)revealed compound
heterozygosity with two novel missense
mutations on PRF1 for both of the
patients. The maternal and paternal muta-
tions are in exon 3, T941G and in exon 2
C46A, respectively; caused the aminoacid
changes of L314R and P16T. Both of them
were novel and missense mutations.
Conclusion: For P1, absence of consan-
guinity, late onset of disease, being trig-
gered by EBV infection and the absence of
any known mutations of PRF1 led us to
consider mainly secondary HLH. However,
5 years later after her death, P2 was diag-
nosed as HLH at 7 months old. Reactiva-
tion of disease during the continuation
therapy and this time, having history of a
sibling death with HLH directed us to a
genetic research for FLH. And, both P1
and P2 had the same novel compound
heterozygous missense mutations. Even
though they had same mutations, P1
became symptomatic at 8 years old, P2 at
7 months old. The clinical and laboratory
ndings, even age of onset and family his-
tory arent only accurate and enough clues
to distinguish FHL and secondary HLH.
New cases of FHL are necessarily needed
to delineate the genotype-phenotype rela-
tion for correct diagnose and effective
treatment.
1445
A pediatric HIV case presented with uni-
lateral parotitis
Hatipoglu, N; Turel, O; Aydogmus, C; Hatipoglu, H;
Babayigit, A; Karabayir, N; Yesil, Y; Siraneci, R
Bakyrkoy Maternity and Childrens Hospital, Pediatrics
Dept., Istanbul, Turkey
Background: Infection due to human
immunodeciency virus (HIV) has been
increasingly reported worldwide and in our
country as well. The diagnosis in children
must be established soon because favorable
prognosis is linked to start treatment early.
Involvement of salivary gland in adults is
not so frequently recognized. We present a
pediatric HIV patient who admitted with
long-lasting unilateral parotid gland swell-
ing.
Case: A four years old boy had been suf-
fering from right parotid swelling and fever
Allergy 66, Suppl. 94 (2011): 482642  2011 John Wiley & Sons A/S
for one month. He was rst diagnosed as
mumps as he was not vaccinated, but the
serology for mumps was found to be nega-
tive. Because of ongoing swelling at the
same side, he was referred to our hospital
for suspected bacterial parotitis, lymphoma
or salivary stone. He had mild anemia (Hb
10.4 g/l), leukocytosis (13 /mm3), thrombo-
cytopenia (54,000 /mm3), and very high
sedimentation rate (120 mm hourly). Mag-
netic resonance of the region showed
enlarged (widest dimension wass 3.5 cm)
right parotid gland and lymphoprolifera-
tive inltration, which was suggestive of
lymphoma or HIV infection. Western-blot
test of the patient was found to be positive.
The family members were unaware of their
HIV status until these results were
obtained, and his mother learned that she
was also infected with HIV when anti-HIV
test was done. The patient was then fol-
lowed up and treated as pediatric HIV
infection.
Conclusion: Children infected with HIV
have a wide range of clinical manifesta-
tions and those with parotid involvement
usually exhibit other ndings. This patient
had no suggestive of HIV disease on
admission other than thrombocytopenia,
which was rather a nonspecic laboratory
work-up for infectious diseases. As HIV
infection is an emerging disease in our
country, one must consider HIV infection
in any case who has unexplained clinical
course.
1446
Clinical and genetic analysis of Turkish
patients with autosomal recessive and
dominant hyper IgE syndrome
Metin, A; Dilek, A; Ozcan, C; Eerkocoglu, M; Akan, A;
Kaya, A; Kocabas, C
Ankara Childrens Hematology Oncology Training
Hospital, Ped Immunology and Allergy Department,
Ankara, Turkey
Background: Background: Hyper IgE syn-
drome (HIES) is a rare primary immuno-
deciency disease, characterized by the
classical triad of chronic pruritic dermatitis
of early onset, severe infections of bacte-
rial, viral, mycobacterial and fungal origin
and elevated levels of serum IgE. HIES
had been shown to be associated with
STAT-3 mutations in AD-HIES patients
and with DOCK8 mutations in AR-HIES.
Notably AR-HIES patients suffer from
severe viral infections such as chronic
refractory molluscum contagiosum, herpes
simplex virus infection, and fatal neurolog-
ical complications which were not usually
observed in AD-HIES.
Case presentations: Here we present 7
HIES patients, 3 of them sporadic or AD-
HIES with conrmed heterozygous domi-
nant negative mutations of STAT-3 gene,
and remaining 4 AR-HIES patients with
conrmed homozygous DOCK-8 gene
mutations. AD-HIES patients presented
with multiple TBC brain abscesses, pneu-
monias with pneumatoceles formation,
multiple cold and hot skin abscesses, sec-
ondary infection of pneumatoceles with
Aspergillus spp., bone fractures with minor
trauma, and non-Hodgkin (T-cell) lym-
phoma. AR-HIES patients were presented
with disseminated HPV infection (epiderm-
odysplasia verruciformis), multiple verruca
plana, melanomas, HSV infections (herpes
labialis, herpes keratitis, herpes hepatitis),
progressive multifocal leucoencephalopathy
due to JC virus infection. However both
types of HIES patients had severe food
allergies which we saw in all of our
patients.
1447
Association of herpes virus infection and
disorders of immune status with inflam-
matory large intestine disease with chil-
dren
Belovolova, R
Rostov State Medical University, Department of Clinical
Immunology and Allergology, Rostov-on-Don, Russian
Federation
Background: The background of the study
was to identify disorders of immune status
and severity of inammation of large intes-
tine and to optimize the therapy.
Methods: The methods used a total of 30
children at the age of 34 years with
chronic colitis were examined. Diagnosis is
based on clinical and laboratory alongside
with endoscopic study. Study of immune
status included: the identication of
peripheral blood lymphocytes by ow
cytometry; serum immunoglobulins, the
level of circulating immune complexes, the
detection of the metabolic activity of
phagocytes, the level of antiviral antibodies
by ELISA.
Results: The results obtained that in 8% of
children with chronic colitis in serum there
were discovered IgG and IgA in the BNA
to the Epstein Barr virus (EBV). 65% of
children - IgG antibodies to CMV. The
simultaneous availability of IgG antibodies
to EBV and CMV were detected in 45% of
patients. Detection of IgM antibodies to
EBV or CMV with the patients is associ-
ated with signicant clinical and/or endo-
scopic signs of acute inammation. An
imbalance of T-and B-lymphocytes was
registered: reduction of CD3 +, CD4 +
lymphocytes, increase of CD20 + lympho-
cytes. Change in metabolic activity of neu-
trophils was manifested through increase in
its spontaneous activity and reduction of
reserved capacity. In 90% of the examined
547

patients levels of serum IgM, 40% of chil-
dren - IgA was increased. In 50% of the
patients the level of CEC was elevated.
The degree of violations of immunological
parameters was combined with the severity
of inammation in the mucosa of the large
intestine.
Conclusion: The conclusion reached inter-
relationship of clinical and immunologic
characteristics and ELISA-based survey
data reects the formation of an inamma-
tory process involving the immune mecha-
nism and participation of herpes virus
infection in the development of inamma-
tion in the lining of the large intestine is
expected. Antiviral drugs (valacyclovir) in
age dosages was included into the complex
treatment of children with chronic colitis
with its worsening combined with detection
by ELISA-based survey of IgM antibodies
to EBV or CMV, and signs of immune sys-
tem activation. Targeted immuncorrection
was carried out with immunomodulators
strictly according to the indication of
immunological analysis. Inclusion of antiv-
iral and immunomodulatory drugs into the
treatment of chronic colitis was accompa-
nied by a good clinical effect, as conrmed
by positive dynamics of immunological
indices.
1448
Immunodeficiency with CD4+ T and
CD19+B lymphopenia with normal immu-
noglobulins in a patient with growth hor-
mone resistant dwarfism
Metin, A1; Demirel, F2; Demirkan, N3; Kacar, A4;
Gunduz, M5; Ozcan, C1; Kocabas, C1
1
Ankara Childrens Hematology Oncology Training
Hospital, Ped. Immunology and Allergy Dept., Ankara,
Turkey; 2Ankara Childrens Hematology Oncology
Training Hospital, Ped. Endocrinology Dept., Ankara,
Turkey; 3Ankara Childrens Hematology Oncology
Training Hospital, Ped. Gastroenterology Dept., Ankara,
Turkey; 4Ankara Childrens Hematology Oncology
Training Hospital, Ped. Pathology Dept., Ankara,
Turkey; 5Ankara Childrens Hematology Oncology
Training Hospital, Ped. Metabolism Dept., Ankara,
Turkey
Case report: 2500 g M was born after a
term pregnancy to a consanguineous cou-
ple. Parents heights were normal There
was no family history of growth failure. At
2 months of age he required hospitalisation
because of respiratory infection. Poor
wight gain and growth failure were noted
all the rst 8 years of life, when he evalu-
ated for the rst time by an endocrinolo-
gist. His height and weight were below the
3rd percentiles. He also had recurrent pul-
548
Poster Session 60 Immunodeciency: the role of T cells
monary infections and respiratory difcul- 1449
ties, and recurrent gastroenteritis leading Possible role of mixed heavy metal toxic-
malabsorbtion. TB, CF test, HIV, CMV, ity in recurrent infections and NK cell
EBV, Hepatitis markers were negative. The count in a population of children with
patient treated with inhaled corticosteroids different forms of immune dysfunction
with partial improvement, At 8 years, he
Luna, G1; Herrera, M1; Benarroch, L2
presented with severe varicella. Progressive 1
Hospital Militar Dr. Carlos Arvelo, Department of
worsening of his pulmonary function Allergy and Immunology, Caracas, Venezuela; 2Hospital
resulted in bilateral bronchiectasis. His de Clinicas, Immunology Pediatric Unit, Caracas,
Venezuela
recurrent chronic supurative otitis media
+ mastoiditis resulted in bilateral sensori-
Background: Developmental immune toxic-
neural hearing loss. At 10 years of age a
ity and immune dysfunction caused by pre-
3 year trial of growth hormone therapy
natal or early childhood environmental
resulted in no improvement. At age
exposures such as infectious agents, xeno-
14.5 years his height was 120 cm with nor-
biotics, maternal smoking, heavy metals,
mal body proportions and delayed second-
antibiotics, alcohol, and drugs, have been
ary sex characteristics. He had a prominent
increasingly recognized as signicant stres-
head with normal head circumference.
sors that could inuence the genetic and
Liver was 4 cm below the right costal ma-
immune outcome in childrens chronic dis-
grin. The patient was referred to our hospi-
eases, We describe a group of children that
tal for further immunological and
share the presence of high levels of heavy
endocrinological evaluation. Serum concen-
metals with immune dysfunction.
trations of IGF-1 and IGFBP-3 were very
Methods: The children were characterized
low which were consistent with the defect
as primary immunodeciencys (PID),
of downstream from GF receptor. Liver
allergies (AD), inborn errors of metabolism
biopsy showed steatosis and chronic hepa-
(IEM) and developmental disorders (DD)
titis with bridging. Hepatic function tests
either by clinical history for heavy metal
showed minimal rise in AST/ALT; GGT,
toxicity or allergy, skin test, metabolic
ALP and LDH were consistent with chole-
analysis, genetic testing, also tested for
stasis. Autoimmune hepatitis, metabolic
DSM IV (diagnostic and statistical manual
diseases excluded as all autoantibody panel
of mental disorders) and CARS (childhood
was negative. Recently some reported cases
autism rating scale). We assessed recurrent
of growth hormone insensitivity involved
infections sites and GI chronic disease. We
mutations of STAT5b gene. These patients
analyzed the immunophenotype and levels
of Stat5b deciency showed low IGF-1
of toxic metals by ow cytometry and hair
and IGFBP-3 levels, no response to GH
sample inductively coupled plasma mass
treatment and some ndings consistent
spectrometry (ICP-MS) respectively.
with immunodeciency as seen in our
Results: We found similar clinical ndings
patient. Only ve patients published to
and Immune dysfunction in all groups.
date have moderately low T cells. Our
Markers of immune dysfunction included
patients immunological workup showed N
Hiper IgE (>2000) poor vaccination
Ig levels, ALC: 6000 /mm3, lymphocytes
response (measles, varicela, rubella, polio).
CD3+ 66% (absolute: 3960 /mm3),
Low NK cells count .High levels of heavy
CD4+: 8% (480 /mm3), CD8+: 55%,
metals (aluminum, lead, mercury, arsenic,
CD19+ and CD20+ B: 2% (120 /mm3).
cadmium) on the overall population, possi-
After 48 h of cell culture with PHA sti-
ble linked to chronic heavy metal toxicity
mul., CD4+CD25+ expression were
exposure.
greatly reduced and CD4+CD45RO+
Conclusions: Our results show an overlap
memory T cell population were 0%, which
of the different forms of immunological
suggested CID. His molecular diagnosis is
dysfunction, with recurrent ORL and GI
ongoing.
tract infections and chronic diseases. The
previously described results suggest that
mixed heavy metal environmental exposure
could be an important factor that has to
be considered as a probably contributor
for the different forms of immunological
dysfunction and could add new insights
and perspectives for their treatment.
Allergy 66, Suppl. 94 (2011): 482642  2011 John Wiley & Sons A/S

The development of allergy and asthma in childhood
1450
Does protection from atopy among pre-
school children from Turkish migrants
persist into adolescence?
Theissen, C; Elingshausen, J; Yozgat, B; Giese, A;
Yurek, F; Wahn, U; Matricardi, P; Gruber, C
Charite Universitatsmedizin Berlin, Pediatric
Pneumology and Immunology, Berlin, Germany
Background: In an earlier study we could
demonstrate that preschool children from
Turkish migrants to Germany are less fre-
quently sensitized to environmental aller-
gens. It is currently unknown whether this
allergy protective effect is transient.
Methods: In 1998, we investigated allergic
sensitization among preschool children
born in Germany and living in an inner-
city district of Berlin, Germany, with
double German or double Turkish parental
citizenship. About 10 years later we started
a re-investigation of this population and
their parents. The preliminary data set cur-
rently contains n = 200 adolescents and
261 parents with blood samples available.
The main outcome measure of the analysis
presented here is specic allergic sensitiza-
tion to common aeroallergens (Phadia-
top 0.35 kU/l). With further examination
in progress, nal data on the investigated
population sample will be available in
spring 2011.
Results: Allergic sensitization was signi-
cantly less prevalent among adolescents of
Turkish origin (n = 98, 30% sensitized)
than among their German peers (n = 102,
46% sensitized; P = 0.016). Correspond-
ingly lower rates of sensitization were
found among Turkish fathers (Turkish
n = 52, German n = 49; 25% versus 48%
sensitized; P = 0.012) and a tendency
towards less frequent sensitization among
Turkish mothers (Turkish n = 74, German
n = 86; 24% versus 33% sensitized;
P = 0.251).
Conclusion: Although the families investi-
gated here live in the same macro-environ-
ment, the migrant subpopulation seems to
be persistently better protected from atopy
than the domestic one. Further study of
this cohort is aimed at investigating the
factors responsible for the observed differ-
ences.
Allergy 66, Suppl. 94 (2011): 482642  2011 John Wiley & Sons A/S
Poster Session 61
1451
Prevalence and risk factors of asthma
among schoolchildren in Gaziantep, a
province in South-Eastern Turkey
Kucukosmanoglu, E1; Palali, M2; Keskin, O1; Kul, S3;
Aydin, N4
1 1
Gaziantep University School of Medicine, Pediatric
Allergy, Gaziantep, Turkey; 2Gaziantep University
School of Medicine, Pediatrics, Gaziantep, Turkey;
3
Gaziantep University School of Medicine, Bioistatistics,
Gaziantep, Turkey; 4Gaziantep University School of
Medicine, Public Health, Gaziantep, Turkey
Background: Asthma is a common chronic
illness in childhood. The aim of this study
was to determine the prevalence of asthma
and its risk factors among the schoolchil-
dren, aged 78 years old in Gaziantep, a
province located in southeast Turkey.
Method: We prepared 2 250 questionnaires
according to International Study of
Asthma and Allergies in Childhood
(ISAAC) criteria and disturbed them in
schools to be lled at home by parents.
Appropriately completed 1 877 question-
naires including complementary questions
for risk factors, were taken into consider-
ation. The response rate was 83.4%.
Result: The cumulative and 12-month
prevalence of wheezing were 28.3 and
11.6% respectively. The prevalence of phy-
sician-diagnosed asthma was 7.0%. Family
history of allergic diseases was the stron-
gest risk factor for ever wheezing (odds
ratio [OR] 2.50, 95% condence inter-
val[CI] 1.983.15). Smoking at home was
the strongest risk factor for current
wheezing (odds ratio [OR] 1.63, 95% con-
dence interval[CI] 1.102.41). History of
pneumonia in the rst two years of life was
the strongest factor for physician-diag-
nosed asthma (odds ratio [OR] 6.00, 95%
condence interval[CI] 3.799.52).
Conclusion: In conclusion, asthma is an
important chronic disease of childhood in
Gaziantep, a province in south-eastern
Turkey.
1452
Associated factors of wheezing at 3 years
of age in Japan: prospective birth cohort
study
Yamamoto, K1; Shoda, T1; Futamura, M1; Narita, M1;
Sakamoto, N2; Aizawa, S3; Saiho, H4; Ohya, Y1
National Center for Child Health and Development,
Department of Medical Subspecialties, Division of
Allergy, Tokyo, Japan; 2National Research Institute for
Child Health & Development, Department of Maternal
and Child Health, Tokyo, Japan; 3National Center for
Child Health and Development, Department of Medical
Information, Tokyo, Japan; 4National Research Institute
for Child Health & Development, Department of Allergy
& Immunology, Tokyo, Japan
Background: Increased prevalence of
asthma and wheezing for these decades
worldwide implies its cause in our lifestyle
changes associated with medical and envi-
ronmental development. The aim of this
study is to explore the cumlative incidence
of wheezing of 3-year-old children and pre-
natal risk factors of their wheezing.
Method: We recruited 1 701 mothers dur-
ing pregnancy in agreement with Tokyo-
Childrens Health, Illness and Develop-
ment (T-CHILD) study. This study sample
consisted of 1 344 mother-infant pairs
from T-CHILD study. Data were obtained
from health event questionnaires during
pregnancy, at 12 months and 3 years of
age. The questionnaires include items of
the International Study of Asthma and
Allergies in Childhood (ISAAC). By logis-
tic regression analysis, we identied the
independent risk factors for wheezing at
3 years of age.
Result: Cumulative incidence of wheezing
at 3 years of age was 24%. The risk factors
for wheezing at 3 years of age were mater-
nal history of asthma (adjusted odds ratio
[aOR], 2.29; 95% CI, 1.533.43), previous
maternal use of oral contraceptives (aOR,
1.89; 95% CI, 1.252.88), breast feeding
(aOR, 1.90; 95% CI, 1.222.97),male gen-
der(aOR, 1.42; 95% CI, 1.041.96), early
daycare attendance (aOR, 1.63; 95% CI,
1.132.36), respiratory infection within the
rst year of life (aOR, 1.56; 95% CI, 1.11
2.10) and bronchitis within the rst year of
life (aOR, 3.37; 95% CI, 2.185.23) respec-
tively. Maternal history of common cold
during pregnancy(aOR, 0.69; 95% CI:
0.480.98) had protective effect against the
development of wheezing at 3 years of age.
Food allergy, atopic dermatitis, pet owner-
ship and maternal smoking during preg-
549

nancy werent signicantly associated with
wheezing at 3 years of age.
Conclusion: Breast feeding and contracep-
tive use before pregnacy were the risk fac-
tors of wheezing
1453
Salivary cortisol and allergy in infants -
results from the Aladdin birth cohort
study
Swartz, J1; Eriksson, M2; Lilja, G2; Bottai, M3; Borres,
M4; Pershagen, G3; Scheynius, A5; Lindblad, F6;
Theorell, T6; Alm, J2; Stenius, F2
1 als and the environmental factors under
Vidar Clinic, Jarna, Sweden; 2Karolinska Institutet,
Dep. of Clinical Science and Education, Stockholm,
Sweden; 3Karolinska Institutet, Institute of
Environmental Medicine, Solna, Sweden; 4Phadia AB,
Uppsala, Sweden; 5Karolinska Institutet, Clinical Allergy
Research Unit, Department of Medicine, Solna,
Sweden; 6Stockholm University, Stress Research
Institute, Stockholm, Sweden
Background: Pre- and postnatal stress has
been related to allergy in children but evi-
dence based on prospective studies is lim-
ited. The aim of this study was to assess
the association between salivary cortisol
levels at six months of age and allergic
manifestations during the rst two years of
life.
Method: Salivary samples for analysis of
cortisol were collected on three occasions
during one day from 198 children in fami-
lies with conventional or anthroposophic
lifestyle at six-month of age. Blood samples
were collected at 6, 12 and 24 months of
age for analyses of specic IgE to common
allergens. Information on allergy related
symptoms was obtained by repeated exam-
inations of the children and interviews.
Result: The adjusted odds ratio (OR) for
the relation between morning cortisol and
sensitisation was 1.59 (95% CI 1.222.09,
P = 0.001) and for eczema 1.28 (95% CI
1.031.59, P = 0.026). The ORs for after-
noon cortisol in relation to sensitisation
and eczema were 1.56 (95% CI 1.21.94,
P < 0.001) and 1.32 (95% CI 1.121.58,
P < 0.001), respectively, and for evening
cortisol 1.49 (95% CI 1.221.83,
P < 0.001) and 1.37 (95% CI 1.181.59,
P < 0.001). Salivary cortisol in the even-
ing was also associated to reported food
allergy.
Conclusion: The association between sali-
vary cortisol levels in infancy and allergic
sensitisation as well as allergy related
symptoms suggests a role of an altered
HPA-axis in the etiological process of
allergic diseases.
550
Poster Session 61 The development of allergy and asthma in childhood
1454
Investigation of filaggrin 501X gene
mutation in Turkish children with atopic
dermatitis and the role of serum immu-
noglobulin levels on atopic dermatitis
phenotypes
Ercan Saricoban, H1; Ozen, A1; Baris, S2; Turgay, I3;
Oztezcan, S4; Cengizlier, M1
1 BeiJing, China
Yeditepe University Faculty of Medicine, Pediatric
Allergy Department, Istanbul, Turkey; 2Marmara
University Faculty of Medicine, Pediatric Allergy
Department, Istanbul, Turkey; 3Yeditepe University
Faculty of Medicine, Department of Medical Biology,
Istanbul, Turkey; 4Yeditepe University Faculty of
Medicine, Department of Biochemistry, Istanbul, Turkey
Background: Atopic dermatitis (AD), the
most common chronic relapsing skin con-
dition of infancy and childhood is a com-
plex multifactorial disease, which arises
from the interaction between strong genetic
and environmental factors. We aimed to
investigate the prevelance and effect of
FLG R501X gene mutation in children
with AD of variable severities. Meanwhile,
serum immunoglobulin (Ig) levels were
assessed to evaluate the role of serum Ig
levels on AD phenotypes, particularly with
an emphasis on the development of asthma
and allergic rhinitis.
Method: We studied 49 children with AD
and 50 age- and sex-matched healthy con-
trols. Children were genotyped for the
mutation in FLG R501X gene. Serum lev-
els of major Ig isotypes were assessed.
Result: Study group consisted of 49
patients (M:26, F:23) with a mean age of
4.9 3.6 years and control group con-
sisted of 50 children (M:30,F:20) with a
mean age of 3.8 2.8 years. Genotyping
of R501X mutation revealed risk alleles in
none of the children in study group or con-
trol group. IgG z-scores were signicantly
higher in patients with AD compared to
controls ( )0.97 1.13 versus )1.48
1.02, P = 0.026). There was a positive
trend in IgG z-scores and a negative trend
in IgA z-scores across the severity of AD.
History of recurrent infections was signi-
acntly associated with asthma and/or AR
(47.8% in patients with asthma/AR versus
3.8% in those without). Children with low
IgG or IgA levels presented at an earlier
age (age at presentation= 1.42 (0.67
5.41) years in patients with low IgA; 4.41
(2.568.35) years in those with normal IgA,
P = 0.08), with lower rates of atopy
(11.1% versus 52.6%, P = 0.030), and mild
type atopic dermatitis (77.8% mild,
P = 0.021).
Conclusion: In a sample of Turkish chil-
dren, FLG R501X genotyping revealed no
risk alleles in variable severities of AD or
healthy controls. Our data suggests that
IgG and IgA levels might have a role in
phenotypic features of AD in terms of
severity and atopic sensitization.
Allergy 66, Suppl. 94 (2011): 482642  2011 John Wiley & Sons A/S
1455
A pilot study: childrens atopy and their
household environment affect the asth-
matic incidence
Zhao, J1; Chen, Q2; He, Q1; Zhang, G1; Bai, J1; Wang, J3
1
Capital Institute of Pediatrics, Asthma Clinic and
Education Center, BeiJing, China; 2BeiJing, China;
3
Chinese Center for Disease Control and Prevention,
Background: Asthma is a chronic allergic
diseases and the prevalence of childhood
asthma increases rapidly in China. A
genetic predisposition of allergic individu-
the combined action creates the result.
Exposure to environmental airborne aller-
gens is considered an important inuence
of the asthma incidence. Though allergens
is widespread and can be contact indoor
and outdoor, but children spend most of
the time at home, the indoor environment
has important implication on the develop-
ment of asthma in children. This research
is to observe and analyze the relationships
among childhood household environmental
factors, their atopy and asthmatic inci-
dence in Beijing urban community, and
nd out the risk factors inuenced to asth-
matic incidence, in order to provide rea-
sonable suggestions for asthmatic
preventional measures in urban commu-
nity.
Method: A total of 144 children diagnosed
with asthma in nearly 1 year from all
schools and kindergartens in sanlitun and
liulitun communities were selected as cases
group. While cluster sampling was done
by classes stratied by grade, 167 samples
not diagnosed asthma or other allergic
diseases composed control group. Skin
picked test was done in tow groups and
the parental environmental questionnaires
were conducted in the two groups, respec-
tively.
Result: The prevalence of positive skin
pricked test (SPT) in case and control group
was 81.7%, 7.84% (ppO2171.548,
P <0.0001); Infantile eczematous history,
rhinitis, family history of allergies, atopy,
textile sofa at home, and mould in toilet
were the risk factors for the asthmatic inci-
dence, that is the asthmatic risk might
increase 1.86416.1683.91346.619
0.8761.178 folds (P < 0.05).
Conclusion: The allergic diseases his-
toryatopy (indicated by positive SPT)
and some household environmental factors
such as indicated textile sofa and toilet
mould have strong correlation with the
asthmatic incidence. The dose-responsive
relationship between the type and quantity
of allergen and asthma need the further
investigation.
Poster Session 61 The development of allergy and asthma in childhood
1456
Does family atopy influence total IgE in
newborns?
De Amici, M1; Spinillo, A2; Bollani, L3; Quaglini, S4;
Perotti, F2; Torre, C1; Licari, A3; Marseglia, G5; Stronati,
M3
1
Foundation IRCCS Policlinic San Matteo, Hospital
Pediatric Clinic, Pavia, Italy; 2Foundation IRCCS
Policlinic San Matteo, Department of Obstetrics and
Ginecology, Pavia, Italy; 3Foundation IRCCS Policlinic
San Matteo, Neonatal Intensive Care Unit, Pavia, Italy;
4
University of Pavia, Dept. Computer Engineering and
Systems Science, Pavia, Italy; 5Foundation IRCCS
Policlinic San Matteo, University Pediatric Department,
Pavia, Italy
Background: Previous studies have indi-
cated that high levels of total maternal IgE
are associated with high levels of total cord
and serum IgE in newborns. However, no
study has investigated the association of a
family history of atopy with total IgE in
newborns. The aim of the study was to
investigate this association.
Method: The study included 526 newborns
and their mothers. Total serum IgE and a
screening test for inhalant allergens and
common food allergens were performed in
mothers. Total serum and cord IgE were
evaluated in newborns. A family question-
naire was developed and specic answers
were selected to identify atopic family
members. Levels of total IgE were detected
by the IFMA procedure (ImmunoCAP
Phadia. Uppsala, Sweden) in peripheral
blood samples from newborns and mothers
and in cord blood samples from newborns.
Concentrations were expressed in kU/l.
Screening tests in maternal serum were
detected with a qualitative assay (Immuno-
CAP Phadia) for graded determination of
IgE antibodies specic for inhalants and
food allergens. Results were expressed as
positive or negative; a cut-off of 0.35 kU/l
for at least one of the two tests was used
to dene maternal atopy. Since test results
can be expressed as a binary variable (posi-
tive/negative), the Chi Square test was per-
formed to investigate the correlation
between maternal/family and newborn
results. A P 0.05 was considered statisti-
cally signicant.
Result: The screening test showed that 223
mothers (42%) were atopic, while family
questionnaires showed that 225 (43%)
newborns were born to an atopic family (ie
atopic mother, father or brother). When
the mother was not atopic, 428 newborns
(81.4%) had a negative total cord IgE and
434 newborns (82.5%) had a negative total
serum IgE. Instead, when the mother was
atopic, 171 newborns (32.5%) had positive
total cord IgE and 122 newborns (23.2%)
had positive total serum IgE. Similar
results were found when we considered
family atopy: when atopy was not present
in the family, 426 newborns (81%) had
negative total cord IgE and 433 newborns
Allergy 66, Suppl. 94 (2011): 482642  2011 John Wiley & Sons A/S
(82.3%) had negative total serum IgE.
When the family was atopic, total cord IgE
was positive in 157 newborns (29.8%) and
total serum IgE was positive in 116 new-
borns (22.1%).
Conclusion: There is evidence that a nega-
tive value for total IgE in newborns is
associated with the absence of family
atopy, but a positive value for total IgE in
newborns was not correlated with family
allergy.
1457
Comparison of total serum and cord IgE
in newborns
De Amici, M1; Spinillo, A2; Bollani, L3; Quaglini, S4;
Perotti, F2; Giunta, V1; Licari, A3; Marseglia, G5;
Stronati, M3
1
Foundation IRCCS Policlinic San Matteo, Hospital
Pediatric Clinic, Pavia, Italy; 2Foundation IRCCS
Policlinic San Matteo, Department of Obstetrics and
Ginecology, Pavia, Italy; 3Foundation IRCCS Policlinic
San Matteo, Neonatal Intensive Care Unit, Pavia, Italy;
4
University of Pavia, Dept. Computer Engineering and
Systems Science, Pavia, Italy; 5Foundation IRCCS
Policlinic San Matteo, University Pediatric Department,
Pavia, Italy
Background: Previous studies have indi-
cated the importance of total serum and
cord IgE detection in newborns as a risk
factor for future allergy development.
However, no study revealed a correlation
in the two measures. The aim of the study
was to investigate such a correlation and
the relationship of total IgE between moth-
ers and newborns.
Method: The study included 518 newborns
and their mothers. Levels of total IgE were
detected by the IFMA procedure (Immu-
noCAP Phadia. Uppsala, Sweden) in
peripheral blood samples from newborns
and mothers and in cord blood samples
from newborns. Quantitative total IgE
concentrations were expressed in kU/l,
according to the traceable calibration to
the 2nd IRP WHO for human IgE. Statis-
tical analysis was performed using the soft-
ware package Medcalc 9 (Frank
Schoonjans, BE). Correlations between
quantitative variables were evaluated by
the Spearmans correlation coefcient (rho).
Since test results can be expressed as a bin-
ary variable (positive/negative), the Chi
Square test was performed to investigate
the correlation between maternal and new-
born results. A p-value 0.05 was consid-
ered statistically signicant.
Result: There was a strong positive corre-
lation (rho = 0.856, P < 0.0001) between
total serum IgE and total cord IgE in new-
borns. Considering the values in terms of
positive/negative results, the Chi Square
test was also highly signicant
(P < 0.0001) and the contingency table
shows a 93.3% concordance of total cord
IgE and Total serum IgE in newborns.
Also, total serum IgE of mothers and new-
borns was highly associated (P = 0.0019;
concordance 77.2%) as well as total cord
IgE and total maternal IgE (P = 0.0002;
concordance 74.9%).
Conclusion: The study showed a signicant
concordance of total serum IgE and total
cord IgE in newborns. Moreover, total
maternal IgE was associated both with
total serum IgE and total cord IgE.
1458
Molecular evaluation of Enterobacteria-
ceae in gut microbiota for infants with
and without eczema
Yap, G1; Chee, K1; Hong, P2; Aw, M1; Shek, P1; Chua,
K1; Liu, W2; Zhang, X3; Leow, D3; Lee, B1
1
National University of Singapore, Paediatrics,
Singapore; 2University of Illinois, United States;
3
Singapore Clinical Research Institute, Singapore
Background: Studies have reported an
association between the prole of fecal
microbiota signatures and the development
eczema. Of these, the Enterobacteriaceae
family were found to be more prevalent in
the fecal microbiota of allergic infants.
This study aims to evaluate and compare
the genus and species of Enterobacteria-
ceae in gut microbiota of infants with and
without eczema by comparison of 16S
rRNA clone libraries.
Method: Infants who are caesarean deliv-
ered and total formula fed at 1 month and
3 month of age were selected as nested case
control study. Fecal samples at 3 months
from 7 selected infants (3 infants with
eczema, 4 healthy infants) were evaluated
by construction of 16S rRNA clone library
for Enterobacteriaceae. Phylogenetic analy-
sis involved RDP Release 10 and MEGA 5
software.
Result: A total of 713 and 670 clones were
obtained for eczema and healthy group
respectively. At genus level, Enterobacter
and Morganella were present exclusively in
healthy infants. Based on phylogenetic tree
constructed at 97% sequences similarity
from both eczema and healthy groups,
Enterobacter aerogenes and Morganella
morganii were present only in healthy
group. However, Citrobacter farmeri was
observed only in eczema group. Escherichia
coli and Klebsiella pneumoniae present in
both eczema and healthy groups.
Conclusion: Unique bacterial groups were
observed to be uniquely associated with
either the eczema or healthy group. These
bacteria targets may directly inuence the
infant immune system resulting in eczema.
These ndings deserve further validation in
larger populations.
551

1459
The impact of eczema on development of
food allergy in childhood: the Japanese
birth cohort (T-CHILD) study
Shoda, T1; Futamura, M1; Yamamoto, K1; Narita, M1;
Sakamoto, N2; Aizawa, S2; Matsumoto, K2; Saito, H2;
Ohya, Y1
1 Jung, H1; Kwon, J2; Lee, S3; Kim, B4; Kim, H5; Yu, J2;
National Center for Child Health and Development,
Division of Allergy, Department of Medical
Subspecialties, Tokyo, Japan; 2National Research
Institute for Child Health and Development, Tokyo,
Japan
Background: Recent studies suggest that
sensitization to food allergens can occur
through environmental allergen exposure
through the eczematous skin. The aim of
this study is to elucidate the relationship
between eczema and development of food
allergy in a birth cohort study.
Methods: Data were obtained from Tokyo
Childrens Health, Illness and Development
(T-CHILD) study which is an ongoing pro-
spective birth cohort study in Japan. The
parents answered questionnaires at the age
of 6 months, 12 months and 3 years in
their children. Independent variables
included parents atopic disease; breast-fed;
introduction of egg and cows milk. The
associations between eczema and develop-
ment of food allergy were analyzed by
using multiple logistic regression analyses.
Results: The numbers of responses were
1 467 at 6 months, 1,368 at 12 months and
1 321 at 3 years of age,respectively. Among
1 266 children without a history of acute
allergic reaction due to food intake at
6 months of age, eczema at 6 months of
age was a risk factor of having food
allergy at 12 months (odds ratio [OR],
5.78; 95% CI, 0.0800.375; P = 0.000)
and at 3 years of age (OR, 3.16; 95% CI,
0.0850.536; P = 0.001), respectively. Sim-
ilarly, among 1 014 children without a his-
tory of acute allergic reaction to food at
12 months of age, eczema at 12 months of
age was a risk factor of having food
allergy at 3 years of age (OR, 5.25; 95%
CI, 0.1240.807; P = 0.016). On the other
hand, among children without a history of
eczema at 6 months of age, food allergy at
6 months was not a risk factor of eczema
neither at 12 months nor at 3 years of age.
Among children without a history of
eczema at 12 months of age, food allergy
at 12 months was not associated with the
risk of eczema at 3 years of age.
Conclusions: Preceding eczema became a
signicant risk of development of food
allergy at the age of 13 years, whereas
food allergy was not risk factor of develop-
ing eczema. This nding implicated the
possibility that food allergy occurred as a
result of sensitization through cutaneous
exposure.
552
Poster Session 61 The development of allergy and asthma in childhood
1460
Correlation between demographic char-
acteristics and indoor allergen sensitisat-
ion among a farming community, a local
community, and an urban region in
South Korea
Hong, S2; Park, K1
1
Presbyterian Medical Center, Department of
Paediatrics, Jeonju, Republic of Korea; 2Asan Medical
Center, Ulsan University, Department of Paediatrics,
Seoul, Republic of Korea; 3Pyeongchon Sacred Heart
Hospital, Department of Paediatrics, Anyang, Republic
of Korea; 4Inje University Haeundae Paik Hospital,
Department of Paediatrics, Busan, Republic of Korea;
5
Inje University Sanggye Paik Hospital, Department of
Paeditrics, Seoul, Republic of Korea
Background: The prevalence of allergic dis-
ease is increasing across the world. The
development of allergic disease is associ-
ated with allergen sensitization. We investi-
gated regional differences of allergen
sensitization and relations between demo-
graphic characteristics and allergen sensiti-
zation among a FC (Farming Community),
a LC (Local Community), and an UR
(Urban Region) in South Korea.
Method: We conducted a cross-sectional
survey of 1 792 children aged 912 years,
from a FC, a LC and an an UR in Korea.
A questionnaire and skin prick test to 16
common allergens were performed.
Result: Participants from FC and LC were
more likely to have pets or livestock, sib-
lings and breast feeding than from UR. A
higher body mass index and a more fre-
quent history of antibiotic therapy during
infancy, and parental allergy were found in
UR. The rate of sensitization to more than
one allergens was higher in UR than FC
and LC. The sensitization rate to cock-
roach was higher in FC and LC than in
UR. The sensitization rate to Alternaria
had a tendency to decrease with urbaniza-
tion. The sensitization rate to house dust
mite and cat were higher in UR than FC
and LC.
Conclusion: The rate and type of allergen
sensitization may differ among regions,
which may be due to environmental or
habitual factors. Therefore, allergen sensiti-
zation can be reduced by controlling these
factors.
1461
Storage mite sensitisation in Turkey
might be related to previous village life
environment
Celebioglu, E; Ozturk, A; Comert, S; Karakaya, G;
Kalyoncu, A
Hacettepe University School of Medicine, Department
of Chest Diseases, Adult Allergy Unit, Ankara, Turkey
Background: SMs (storage mites) are
known to cause allergic respiratory symp-
toms after occupational sensitization in
Allergy 66, Suppl. 94 (2011): 482642  2011 John Wiley & Sons A/S
farms, dairy barns, bakeries and grain ele-
vator stores. Recently, they also have been
shown in house dust and sensitization rate
to SMs have been shown to be high. We
aimed to investigate if past and current liv-
ing conditions inuence the risk of SM
sensitization in Turkey.
Method: A total of 321 patients (70%
females) aged 33.6 11.9 (1468) years,
were studied in our allergy unit from Sep-
tember 2009 to December 2010. Patients
diagnosed with perennial or intermittent
rhinitis and/or asthma were included in the
study. Skin prick tests to SMs (Lepidogly-
phus destructor, Tyrophagus putrescentiae,
Acarus siro), pollens, moulds and animal
danders were performed in all cases.
Wheals 3 mm were considered positive.
Epidemiological data concerning demo-
graphic data and characteristics of the house
the patient lived (current house and patient
lived until 2 years old) were assessed by
questionnaire, lled out by the physician.
Result: A total of 102 (31.8%) patients
were sensitized to at least one of the SMs
and among SM positive patients 43.1%
were also sensitized to Dermatophagoides
pteronyssinus (P < 0.0001). SM sensitiza-
tion was not signicantly associated with
sex, age, smoking and education level.
When current and past living conditions
were compared, living in inland or coastal
region of Turkey, presence of pets, heating
condition of the houses, type of oor cover-
ing were also not signicantly associated
with SM sensitization. Past and current
house was a country house in 30.5% and
1.9% of patients, respectively and living in
a country house during the rst years of life
was associated with SM sensitization (P:
0.031). Current living conditions were not
signicantly associated with any variable.
Conclusion: Turkey is a developing country
and living conditions have been changing.
Concerning our country, SM sensitization
might be a reection of past village life.
Although SMs are currently isolated from
house dust samples, sensitization to them
may not be related to current disease and
relevance of this sensitization to disease
itself should further be investigated.
1462
The effect of healthcare associated infec-
tions and broad spectrum antibiotic use
in newborn period on development of
asthma, allergic rhinitis and atopic der-
matitis in early childhood
Yapicioglu, H1; Ozcan, A1; Yilmaz, M2; Kendirli, S1;
Altintas, D1; Bingol-Karakoc, G1
1
Turkey; 2Cukurova University Hospital, Pediatrik
Allergy and Immunology, Adana, Turkey
Background: Epidemiologic studies have
demonstrated a signicant increase of
Poster Session 61 The development of allergy and asthma in childhood
allergic diseases over the past decades. It
has been reported that the lack of early
childhood infection agents modulates the
immune system, increasing the susceptibil-
ity of individuals to atopic diseases, partic-
ularly asthma. There are conicting results
about the relationship between antibiotic
use in early childhood and the develop-
ment of atopic diseases. Aim: to investigate
the effect of healthcare associated infec-
tions (HAIs) and broad spectrum antibiotic
use in newborn period on asthma, allergic
rhinitis and atopic dermatitis.
Method: A total of 73 children treated for
HAIs in newborn period in Neonatal Inte-
sive Care Unit between January 2001 and
September 2007, and their 41 siblings who
were healthy in newborn period were
included in the study. Parents answered a
detailed questionnaire, children were exam-
ined and eosinophil count, serum total Ig
E and specic Ig E levels were studied.
Result: Ventilator associated pneumonia
(VAP) was observed in 32 (45.2%), blood
stream infection (BSI) in 28 (38.4%) and
clinic sepsis in 12 (16.4%) of 73 children
with HAIs. 27 (23.6%) of 114 subjects had
at least one atopic disease or asthma. 24 of
these children were from HAI group.
Asthma was signicantly higher in HAIs
group compared to sibling group (32.9%
versus 4.9%, P = 0.001), whereas there
was no signicant difference in allergic rhi-
nitis (4.1% versus 2.4%, P = 0.999) and
atopic dermatitis (6.8% versus 2.4%,
P = 0.417) among groups. Major risk fac-
tors for an allergic disease were healthcare-
associated infections in newborn period,
high C/S delivery rate, lower age, frequent
RTI and hospitalisation for RTI in 1
12 months.
Conclusion: Asthma was signicantly
higher in healthcare associated infections
group, and allergic disease risk seems to
increase in children treated with broad-
spectrum antibiotics for healthcare associ-
ated infections in newborn period.
1463
Role of exposure to risk factors during
the first life year in the development of
respiratory allergic disease
Xhixha, F1; Bakiri, A2; Hyso, E3; Gjata, E4; Mingomataj,
E5; Elezi, K1; Denizi, V1
1
Specialties Policlinic No 3, Allergology Unit, Tirana,
Albania; 2University of Tirana, Nursing Faculty,
Preclinical Specialties, Albania; 3Regional Hospital of
Vlora, Allergology Unit, Albania; 4District Hospital of
Lushnja, Allergology Unit, Albania; 5Mother Theresa
School of Medicine, Dept. of Allergology & Clinical
Immunology, Tirana, Albania
Background: Different environmental fac-
tors are accused as risk factors in the
development of respiratory allergic disease
(RAD) like allergic rhinitis and bronchial
Allergy 66, Suppl. 94 (2011): 482642  2011 John Wiley & Sons A/S
asthma. This study aimed to investigate the
role of exposure to suspected risk factors
during the rst life year (FLY) in the RAD
development during childhood.
Method: We collected the parental reports
during the period 20082010 about their
respiratory atopic children (276, m 160, f
116, age 114 years old, mean age 5.6)
regarding conduction of cesarean section,
breast feeding, exposure to high house
humidity, pets, or to passive tobacco
smoke, and presence of familiar atopy or
atopic dermatitis during the FLY. These
results were compared with collected nd-
ings about a sample of non-allergic chil-
dren (37, m 21, f 16, mean age 5.9),
diagnosed with common upper respiratory
infections. With respect to age and sex, the
control sample data matched with study
contingent ones. The chi2-test was used to
compare the groups data.
Results: Conduction of cesarean section
(46/276 versus 1/37, P < 0.04), exposure to
high humidity at home (164/276 versus 10/
37, P < 0.001), or to passive tobacco
smoke (105/276 versus 5/37, P < 0.005), as
well as presence of atopic dermatitis (67/
276 versus 1/37, P < 0.004) during the
FLY was signicantly associated with
RAD development. Similar tendency was
observed regarding familiar atopy (38/276
versus 1/37, P < 0.06). Exposure to pets
resulted insignicant (43/276 versus 4/37,
P < 0.5), while breast feeding resulted a
non-signicant protective factor (194/276
versus 29/37, P > 0.3).
Conclusions: Cesarean section during birth,
increased humidity level at home, atopy
and passive tobacco smoke during the
FLY could be risk factors regarding the
RAD development. The avoidance of these
factors at least during the mentioned per-
iod may reduce the development risk for
these pathologies.
1464
Early life risk factors for asthma in
Moldavian children
Savoschin, D; Cirstea, O; Cojocaru, A; Vasilos, L;
Horodisteanu-Banuh, A; Livitchi, L
Research Institute for Maternal and Child Health Care,
Scientific Department of Pediatrics, Chisinau, Republic
of Moldova
Background: Epidemiologic investigations
have provided valuable insight into the role
of genetic and environmental factors on
wheezing illnesses in children. This study
was designed to evaluate the most impor-
tant risk factors in infancy and asthma
onset in Moldavian children.
Method: A case-control study was nested
within the cross-sectional study of 789 chil-
dren from the Republic of Moldova,
between January 2007 and May 2010.
Ninety children (51 males and 39 females)
with asthma and 90 matched healthy con-
trols (age range 517 years old) were com-
pared. General characteristics including
detailed information about family history
of asthma and allergy, feeding practices in
infancy and household environment were
recorded by a structured interview. Also,
the association between the risk of early-
onset asthma (up to and including age 3)
and viral infections during infancy was
assessed.
Result: Children with family or maternal
history of allergy were at 3.6-fold higher
risk of asthma than those who were not
[OR = 3.59; 95% condence interval
(95% CI), 1.847.03]. We found also that
compared with children who were breast-
fed, those who were not had a 3.5 times
higher risk of developing childhood asthma
(OR = 3.50; 95% CI, 1.836.68). Early-
onset asthma was present in 61.1% sub-
jects and late-onset form of the disease in
38.9%. In subgroup analysis, viral infec-
tions during infancy was positively associ-
ated with an increased risk of subsequent
wheezing and early-onset asthma
(OR = 3.77; 95% CI, 1.469.74). Further-
more, the OR for the interaction between
the respiratory virus infection during
infancy and history of maternal allergy or
asthma was statistically signicant
(OR = 5.4; 95% CI, 0.8235.65). No sig-
nicant differences in the associations of
viral infections with early-life asthma in
children by breast-feeding were found.
Conclusion: The early life events play a
critical role in the pathogenesis of asthma.
Though the cohort was small, the evidence
from this study points to the likelihood of
increased risk of early-life asthma among
genetically susceptible children who have a
history of viral respiratory infections dur-
ing infancy. It is important to investigate
the relationship between patterns found
and environmental data to identify factors
that may relate to these ndings.
1465
Antibiotic and antipyretic use in first
years of life and development of symp-
toms of allergy diseases in Croatian chil-
dren
Munivrana Skvorc, H1; Plavec, D1; Munivrana, S2;
Skvorc, M3; Nogalo, B4; Turkalj, M1
1
Childrens Hospital Srebrnjak, Department of allergy
and pulmology, Zagreb, Croatia; 2County Hospital
Cakovec, Department of pediatrics, Cakovec, Croatia;
3
County Hospital Cakovec, Department of gynecology,
Cakovec, Croatia; 4Childrens Hospital Srebrnjak,
Hospital Manager, Zagreb, Croatia
Objective: To estimate the association
between the use of antibiotics in the rst
three years of life and use of antipyretic
drugs in the rst year of life and the
553

development of symptoms of asthma and
allergic diseases in the Medjimurje-Cako-
vec County in Northwest Croatia.
Subjects and methods: The study was
undertaken during the 2007/08 school year
among the 1012-year-old school children.
Data were collected using standardized
ISAAC phase II written questionnaire
completed by parents. Exposure to antibi-
otics was documented separately for the
rst, second and third year of life and for
antipyretics for the rst year only.
Results: A total of 600 children partici-
pated in the study (300 with symptoms of
allergic diseases). The use of antibiotics
during the rst; and second and third year
of life was signicantly associated with
wheeze ever (2 = 13.984, P = 0.007;
2 = 24.331, P < 0.001), but not with
symptoms of allergic diseases during last
12 months except for a marginally signi-
cant association of the use of antibiotics in
the rst year of life and allergic rhinitis
(2 = 9.467, P = 0.05). The use of antipy-
retics during the rst year of life was asso-
ciated with an increased risk of wheeze
ever (2 = 9.776, P = 0.008), together
with an increased risk of symptoms of
allergic rhinitis (2 = 16.647, P < 0.001)
and atopic dermatitis (2 = 6.822,
P = 0.033) during last 12 months but not
with symptoms of asthma.
Conclusions: Our ndings do not support
the hypothesis that antibiotic use in early
life is associated with the development of
symptoms of asthma and allergic diseases in
childhood although we found a signicant
association between their uses and wheeze
ever showing that respiratory tract infec-
tions in young children may be confused
with early symptoms of asthma. Use of anti-
pyretic drugs in the rst year of life could
enhance the risk for allergic rhinitis and ato-
pic dermatitis at age 1012 years thus show-
ing that exposure to antipyretic drugs might
be associated with risk for the development
of allergic diseases in childhood.
1466
Children exposure to allergenic mites in
dwellings and childcare facilities
Niesler, A1; Solarz, K2; Gorny, R3; Zlotkowska, R4;
Kasznia-Kocot, J4; Scigala, G1; Ludzen-Izbinska, B1;
Sakol, G1; Brewczynski, P1
1
Institute of Occupational Medicine and Environmental
Health, Department of Biohazards and
Immunoallergology, Sosnowiec, Poland; 2Medical
University of Silesia, Department of Parasitology,
Sosnowiec, Poland; 3Central Institute for Labour
Protection National Research Institute, Department of
Chemical and Aerosol Hazards, Warsaw, Poland;
4
Institute of Occupational Medicine and Environmental
Health, Department of Environmental Health and
Epidemiology, Sosnowiec, Poland
Background: Many mite species are the
important source of allergens in indoor
554
Poster Session 61 The development of allergy and asthma in childhood
environment. They may cause the sensitiza-
tion and development of some allergic dis-
eases. Children are the most sensitive
group. Considering the scale of this phe-
nomenon, the aim of this study was to
asses children exposure to allergenic mites
in their homes and childcare facilities.
Method: The study was carried out in 16
kindergartens, 3-day care centers and 17
dwellings located in the Upper Silesia
(Poland). Quantitative and qualitative
analyses of mites were performed in dust
samples as well as Der p 1 allergen concen-
trations were measured in settled dust and
ambient air. Parents of 736 children
answered a standardized questionnaire. In
serum of pre-school children with symp-
toms of allergy to mites the levels of spe-
cic IgE to Der p1 were determined.
Result: The concentrations of Der p 1
allergen in settled dust ranged from <0.1
to10.18 lg/g in childcare facilities and from
<0.1 to 11.31 lg/g in dwellings, whereas
the densities of mites ranged from 0 to 833
and from 0 to 5434 per 1 g of dust, respec-
tively. Der p 1 allergen concentrations in
ambient air didnt exceed the level of
10 ng/m3. The highest Der p 1 concentra-
tions and mite densities were observed in
dust samples taken from soft toys in child-
care facilities and childrens beds in dwell-
ings. Allergenic species belonged to
Pyroglyphidae family constituted >95% of
all collected mites and Dermatophagoides
pteronyssinus was predominant. During
questionnaire survey, a total number of
104 children with suspected allergy to mites
were identied. In such children, the preva-
lence of positive specic IgE (>0.35 KU/l)
was 22.4%.
Conclusion: Children exposure to allergenic
mites in the studied childcare facilities was
low. In all kindergartens and day care cen-
ters Der p 1 allergen mean concentration
as well as mean density of mites (except
for one nursery school) were below the
level which is directly attributable to the
increased risk of sensitization (2 lg/g and
100 mites/g, respectively). Signicantly
higher concentrations were observed in
childrens homes, compared with the child-
care facilities. There were not any correla-
tions between the density of mites, Der p 1
concentration and the level of specic IgE.
Allergy 66, Suppl. 94 (2011): 482642  2011 John Wiley & Sons A/S
1467
Influence of environmental chemicals and
drugs taken before and during pregnancy
on onset of childhood asthma and
eczema
Ohya, Y1; Yonemoto, J2; Ogata, T1; Sone, H2;
Sakamoto, N1; Qiu, D1; Yamamoto, K1; Shoda, T1;
Futamura, M1; Narita, M1; Tsumura, Y1; Hamaguchi,
M1; Doi, M1; Horikawa, R1; Aizawa, S1; Kitagawa, M1;
Saito, H1
1
National Center for Child Health and Development,
Tokyo, Japan; 2National Institute for Environmental
Studies, Tsukuba, Japan
Background: Although maternal intake of
drugs and foods during pregnancy have
been thought to have some effects on the
onset of offsprings allergic diseases, scarce
reports were seen so far except those focus-
ing on antibiotics, probiotics, prebiotics
and vitamins. The aim of this study is to
explore the inuence of prenatal exposure
of environmental chemicals contained in
foods and drugs taken by pregnant women
on postnatal allergic manifestations such as
wheezing and eczema.
Method: Maternal history of drug intake
during pregnancy and their childrens dis-
ease histories were extracted from database
of a birth cohort study named Tokyo Chil-
drens Health, Illness and Development (T-
CHILD). Environmental chemicals were
detected from umbilical cords of partici-
pants who agreed to provide a part of
them to this study.
Result: Doctor diagnosed asthma at the
age of three years increased its risk when
maternal history of contraceptive intake
was positive (OR:1.84, 95% CI 1.282.62),
but wheezing at the age of two years did
not. Maternal contraceptive use was not
risk factor of doctor diagnosed eczema at
the age of two and three years in their chil-
dren. Iron preparation and folic acid sup-
plement during pregnancy did not show
positive association with asthma and eczema
of their offspring at two and three years of
age. Among 44 kinds of environmental
chemicals assayed in this study, 4-OH-
2,23,4,5,5-hexachlorobiphenyl (4M146)
showed higher concentration in umbilical
cords of two years old children with atopic
eczema than in those of children without
eczema (77.5 versus 60.4 pg/g-fat, P < 0.05)
and 4-OH-2,2,4,5,5-pentachlorobiphenyl
(4PM101) showed higher concentration in
umbilical cords of 3 years old children with
wheezing episode than in those of children
without wheezing history (32.7 versus
21.3 pg/g-fat).
Conclusion: Some of environmental chemi-
cals and drugs which have epigenetic inu-
ence on maternal body may become risk
factors to increase asthma and/or eczema
onset of offsprings.

Pediatric allergy: assessing the burden of disease
1468
The association between common men-
tal disorders, sleep problems and quality
of life in adolescents with asthma
Nogueira, K1; Lopes, C2; Johanna, P3
1
State University of Rio de Janeiro, Center of Study of
Adolescent Health, Rio de Janeiro, Brazil; 2State
University of Rio de Janeiro, Brazil; 3Brazil
Background: Asthma is the most prevalent
chronic disease among adolescents, affect-
ing their quality of life and raising con-
cerns about their health. Beyond the
limitations imposed by the disease itself,
any chronic disease in this age range
increases a sense of vulnerability and has
emotional consequences including common
mental disorders (CMD). Objective: Evalu-
ate the association between CMD and
quality of life and sleep in adolescents with
asthma.
Method: This transverse study investigated
210 adolescents with asthma between ages
12 and 21, seen at a university outpatient
adolescent medicine service in Rio de
Janeiro, Brazil.. The Paediatric Asthma
Quality of Life Questionnaire (PAQLQ)
and General Health Questionnaire (GHQ-
12) were used to assess QoL and common
mental disorders (CMD) respectively. Total
quality of life and its various dimensions
were treated as dichotomous variables. A
binomial log-rithmic model was used to
calculate raw and adjusted prevalence
ratios.
Result: The prevalence of asthmatics with
CMD was 32.4%. And asthmatic with
sleep problems was 28%. Examining
CMDs relation to QoL, the prevalence of
poor QoL was 36.6%. And with tree dis-
eases was 24% The adjusted nal models
showed an association between CMD and
poor total quality of life (PR = 1.84 IC
95% 1.192.86) as for the domains related
to emotions (PR = 1.77 IC 95% 1.16
2.62) and symptoms (RP = 1.75 IC 95%
1.142.70). For the domain related to
physical activity, the association to CMD
was only borderline (RP = 1.43 IC 95%
0.972.72).
Conclusion: The results of this study sug-
gest that greater attention be paid to the
emotional needs of adolescents with
chronic diseases, and also with their qual-
ity of sleep including more effective initia-
tives in the area of mental health in order
Allergy 66, Suppl. 94 (2011): 482642  2011 John Wiley & Sons A/S
Poster Session 62
to promote better quality of life and more
comprehensive treatment for young
patients with asthma.
1469
Impact of food allergy on the quality of
life in Swiss children
Wassenberg, J1; Cochard, M2; DunnGalvin, A3;
Ballabeni, P4; Flokstra-de Blok, B5; Hofer, M1;
Eigenmann, P2
1
University Hospital of Lausanne, Division of
Allergology, Immunology and Rhumatology,
Department of Paediatrics, Lausanne, Switzerland;
2
University Hospital of Geneva, Allergology,
Department of Paediatrics, Geneva, Switzerland;
3
University College of Cork, Department of Paediatrics
and Child Health, Cork, Ireland; 4University Hospital of
Lausanne, Division of Epidemiology, Lausanne,
Switzerland; 5University Medical Center of Groningen,
Department of General Practice, Groningen,
Netherlands
Background: Food allergy in children, an
increasingly prevalent disease, signicantly
affects the quality of life. Its impact can be
analyzed by the validated French versions
of the Food Allergy Quality of Life Ques-
tionnaires, parents form and childrens
form (FAQLQ-PF and FAQLQ-CF).
Objectives: The aim of our study was to
evaluate the quality of life in a sample of
Swiss children with IgE-mediated food
allergy, conrmed by a standardized food
challenge, or a convincing history of severe
systemic reaction and positive allergy tests.
Methods: Information were collected with
the FAQLQ-PF questionnaires (parents of
children 012 years old) and the FAQLQ-
CF questionnaire (children of 812 years
old) during a scheduled allergy clinic visit,
and analysed in term of emotional impact,
food anxiety and social and food limita-
tions. Patients were divided according to
the FAQLQ-PF questionnaire in three age
groups: group 1 from 0 to 3 years, group 2
from 4 to 6 years and group 3 from 7 up
to 12 years.
Results: A total of 62 food allergic patients
were included, with a girl/boy ratio of
1:1.38. Median age was 7.8 years. 58%
were known to be allergic to one food,
18% to two foods, 24% to 3 or more
foods. Peanuts (71% of children) as well as
hazelnuts (29%), walnuts (23%) and eggs
(22%) were the major allergens. 61% of
children suffered from a mild systemic
reaction due to food allergy, 39% from a
severe systemic reaction, with slightly less
severe reactions (statistically not signi-
cant) among older children. Young chil-
dren (group 1) showed signicantly better
quality of life than older children (group 2
and 3), (Global score (GS) = 0.66 versus
2.13; P = 0.02). Children suffering from
severe systemic reactions showed worse
scores than the ones with mild reactions
(GS = 2.32 versus 1.47; P = 0.039). Chil-
dren with an allergic mother showed worse
scores (GS: 2.22 versus GS = 1.29;
P = 0.002), as the ones with allergic sib-
lings (emotional impact score = 2.12 ver-
sus 1.38 P = 0.03). Multiple regression
calculations could show that these vari-
ables were independent. The type of food
allergens has no signicant impact on qual-
ity of life.
Conclusion: Food allergy affects the qual-
ity of life of Swiss children. Older children,
children with severe systemic reactions and
children with mothers or siblings also
affected by allergies show worse quality of
life scores.
1470
Children with food allergy: validation of
the French version of the food allergy
quality of life questionnaire - children
form
Wassenberg, J1; Cochard, M2; Flokstra-de Blok, B3;
DunnGalvin, A4; Ballabeni, P5; Newman, C6; Hofer, M1;
Eigenmann, P2
1
University Hospital of Lausanne, Division of
Allergology, Immunology and Rhumatology,
Department of Paediatrics, Lausanne, Switzerland;
2
University Hospital of Geneva, Allergology,
Department of Paediatrics, Geneva, Switzerland;
3
University Medical Center of Groningen, Department
of General Practice, Groningen, Netherlands;
4
University College of Cork, Department of Paediatrics
and Child Health, Cork, Ireland; 5University Hospital of
Lausanne, Division of Epidemiology, Lausanne,
Switzerland; 6University Hospital of Lausanne,
Department of Paediatrics, Lausanne, Switzerland
Background: It has been previously shown
with English speaking children that food
allergy clearly affects their quality of life.
A health-related quality of life question-
naire for food allergic children has been
validated in Dutch in 2009, however to
date no such questionnaire is available in
French.
Objectives: To validate the French version
of the Food Allergy Quality of Life Ques-
tionnaire- Children Form (FAQLQ-CF)
555

developed and validated in Dutch and
available in English.
Methods: The English version of the FA-
QLQ-CF was translated to French by two
different French-speaking translators and
retranslated by a English-speaking transla-
tor. We then recruited 32 patients between
8 and 12 years with an IgE mediated food
allergy. Children answered the question-
naire during a regular clinic visit. The
results obtained were then analysed and
compared with the results of the original
FAQLQ-CF and the Food Allergy inde-
pendent Measure (FAIM).
Results: 32 questionnaires were fully com-
pleted and available for analysis. A Cron-
bachs a correlation index of 0.67 was
found for the total FAQLQ-CF. Validity
was demonstrated by signicant correla-
tions between FAQLQ-CF and the FAIM
(0.67, P < 0.05).
Conclusion: The French version of the FA-
QLQ-CF was validated and will provide a
tool for assessment of Quality of Life in
French-speaking children with food allergy.
It will be an important tool for clinical
research and will allow research collabora-
tion between French and English speaking
research teams.
1471
The effects of educational intervention
on childhood allergy on primary care
physicians
Niculescu, C1; Gheonea, C1; Oprica Rusoiu, C2
1 broad understanding of WHB project
University of Medicine and Pharmacy of Craiova,
Department of Paediatrics, Craiova, Romania;
2
Emergency Hospital Craiova, Department of
Allergology, Craiova, Romania
Objective: To determine baseline knowl-
edge of primary care physicians on the sys-
temic nature of childhood allergy, and to
assess the inuence of an educational event.
Method: One hundred sixty-four primary
care physicians who attended an education
symposium were enrolled. All primary care
physicians received a structured question-
naire at the beginning and at the end of
the intervention. The topics addressed were
on the systemic nature of childhood aller-
gies and the atopic march.
Result: Mean age of the primary care phy-
sicians was 40.2 11.4 years. Mean dura-
tion after graduation from medical faculty
was 16.1 9.2 years. There was signicant
improvement in answers after education.
Systemic nature of allergy was approved
by 62%, which increased to 98% after the
education (P < 0.001).
Conclusion: Although some primary care
physicians may lack updated information
about the systemic nature of paediatric
allergic diseases, a single educational event
may improve their knowledge signicantly.
556
Poster Session 62 Pediatric allergy: assessing the burden of disease
1472
Parents, pediatricians, and pharmacists.
Do they speak the same language? A
focus on pediatric allergy
Al-Dqour, H
MSD, Merck & Co., Inc., Global Human Health, Amman,
Jordan
Background: Is it the culture, we live, that
directs the science, we practice? Or, is it
the science that shapes the culture? Contro-
versial question! What I am sure of; is that
we should formulate and execute the best
of methods to attain a bridge that we will
live on. Pediatric allergy considered as one
of the challenging clinical issues, especially
the multiple inter-crossing points for deci-
sion making process in treatment plan. In
a dynamic environment, like the medicine
practice nowadays, the urgency for a glo-
bal customer relationship management
(CRM) system brings it all together is
raised.
Method: Descriptive in nature; through
this paper an attempt has been made to
correlate the world health bank (WHB)
functionality through a Ps triangle model
that can be used in the pediatric allergy
settings.
Result: Provide a denition of Ps triangle;
which is a plot represents the relationship
between the major partners concerned with
pediatric allergy as been correlated to
WHB. This gure illustrates the potential
access points that (WHB) works on. Ps tri-
angle is fundamental approach to build a
among healthcare partners.
Conclusion: Ps triangle provides a unique
opportunity to integrate the partners
effort for a successful treatment plan for
pediatric allergy.
1473
Allergic pathology in pediatric patients
from the Puertollano health area (Ciudad
Real, Spain)
Alfaya, T1; Mur, P1; Martn, A1; Rodriguez, D2
1
Santa Barbara Hospital, Allergy Unit, Puertollano,
Spain; 2Diater Laboratory, I + D department, Madrid,
Spain
Background: The Puertollano Allergy Unit
covers a pediatric population of 10945 chil-
dren. 964 pediatric patients were attended
in the allergy consultation between 2006
and 2008. In this study we analyze the
allergic pathologies diagnosed in this popu-
lation, and we compare these data with
those from the national allergy study Al-
ergologica 2005.
Method: We performed a retrospective,
descriptive, transversal study including
data of diagnosis, age, sex, residence and
attendance year. We included the patients
Allergy 66, Suppl. 94 (2011): 482642  2011 John Wiley & Sons A/S
rst attended in the Allergy Unit between
the years 2006 and 2008.
Result: We included 964 patients, with a
mean age of 7.22 years. There was a pre-
dominance of boys (58.2%) over girls
(41.8%). The main diagnosed pathologies
were asthma (46.68%), rhinitis (45.44%),
drug allergy (10.27%), and atopic dermati-
tis (8.20%). Comparing our data with Al-
ergologica 2005, we found signicantly
more diagnosis of asthma (46.68% versus
40.46%) and drug allergy (10.27% versus
7.52%), and less diagnosis of food allergy
(7.26% versus 14.5%) and atopic dermati-
tis (8.20% versus 11.56%). We also found
signicant differences in the rhinitis and
asthma etiology, with more diagnosis of
non allergic rhinitis (21.15% versus 14%)
and non allergic asthma (46.89% versus
18%). The main allergen causing allergic
rhinitis and asthma was pollen with a clear
predominance over other allergens.
Conclusion: The main diagnosis in the
pediatric patients attended in the Puertol-
lano Allergy Unit between 2006 and 2008
were asthma, rhinitis, drug allergy and ato-
pic dermatitis. We found a signicant
increase of non allergic rhinitis and asthma
compared with the national data according
to the Alergologica 2005 study, which
could be due to an early derivation from
pediatricians. The predominance of pollen
over other allergens causing allergic rhinitis
and asthma is explained by the dry conti-
nental climate in our area. The increase in
the diagnosis of drug allergy may be due
to a massive derivation because our drug
allergy unit started in 2006. We found few
diagnosis of food allergy, probably due to
a poor derivation from pediatricians who
initially treat this pathology by themselves.
1474
ISAAC findings in Georgia
Abramidze, T1; Rukhadze, M2; Gotua, M2; Gamkrelidze,
A2
1
Center of Allergy & Immunology, Immunology, Tbilisi,
Georgia; 2Center of Allergy & Immunology, Allergy,
Tbilisi, Georgia
Background: Georgia, which former was
one of the Soviet Union countries, gained
its independence in 1990 and faced most
complicated political situations and hard
economic conditions. The lack of any epi-
demiological data regarding markers of
allergy diseases for that time in Georgia as
well as the interest to conrm the purpose
that the prevalence of these diseases in
Georgia should be much lower than in
country with market economy (as it was
revealed in the other less industrialized for-
merly socialist countries) were the main
reasons of participation of Georgia in this
global epidemiology study.
Poster Session 62 Pediatric allergy: assessing the burden of disease
Method: All three phases of ISAAC was
performed strictly according to the stan-
dardized methodology. ISAAC Phase I (13
516 participants) and ISAAC Phase III (11
655 participants) were conducted in two
cities of Georgia-Tbilisi and Kutaisi, in
19961995 and 2003 correspondently.
ISAAC Phase II (1036 participants) was
performed in Tbilisi, in 20012002.
Results: The prevalence of symptoms of
allergic diseases was less than 5%. The dif-
ferences between two study centers in
symptoms were not obvious among 13
14 years old children. The prevalence of
wheezing and conjunctivitis were slightly
higher in Kutaisi than in Tbilisi among the
67-year olds children. The prevalence rate
of asthma in ISAAC phase II became
9.2%, the prevalence rate of itchy rash and
exural dermatitis were 7.6% and 5.9%,
respectively. The prevalence of current rhi-
noconjunctivitis was increased (6.3% versus
4.7%). The prevalence changes, particu-
larly the increasing pattern, more clearly
expressed in Tbilisi than in Kutaisi centre.
Among adolescences in Kutaisi center only
current wheezing increased and the prev-
alence of all other allergic diseases
decreased or remained without changes.
The prevalence of current wheezing among
67 years old children decreased by 2.4%,
the symptoms of current rhino-conjunctivi-
tis slightly decreased and symptoms of ex-
ural dermatitis reduced by 2.8%.
Conclusion: The prevalence of symptoms of
allergic diseases in Georgia according to the
results of ISAAC ranks as a lowest. The
family history of allergic diseases and damp
spots on the wall was the main determi-
nants for all types of allergic symptoms.
The results of ISAAC phase III study indi-
cate that the epidemiological features of
asthma and allergies in Georgia are chang-
ing, although the causes are still uncertain.
1475
Epidemiological data of a pediatric
allergy outpatients clinic in the meditte-
ranean island of Crete, Greece
Stefanaki, E; Aggelakou, V; Kokori, E; Tsilimigaki, A
Venizelion General Hospital, Pediatric Allergy
Outpatients Clinic, Heraklion, Greece
Background: The island of Crete, 600.000
populations, is geographically located in
the eastern Mediterranean area. Heraklion,
the largest town along the north coast,is
characterized by a mild temperate climate.
Our outpatients pediatric allergy outpa-
tients clinic is run twice a week and most
of our patients are referrals from primary
care pediatricians.
Method: We studied retrospectively the
charts of the patients examined between
Allergy 66, Suppl. 94 (2011): 482642  2011 John Wiley & Sons A/S
December 2009-December 2010.Most of
these patients were followed up at least
twice during this year. Allergy evaluation
included either skin prick tests or specic
IgEs (CAP) to the main aeroallergens or
the implemented food allergens.
Result: We followed-up 346 patients with
median age 6.5 years. 171 (49.4%) of the
children were diagnosed with asthma .91
(53.2%) of them were sensitized to aeroal-
lergens but 13 of these sensitized individu-
als had only viral triggered episodes. Most
sensitized children with asthma (78%) had
also allergic rhinitis. 106 (30%) of all chil-
dren had allergic rhinitis but in only 10%
of them rhinitis is their only allergic dis-
ease. Considering the prevalence of aeroal-
lergens in our 147 sensitized patients, dust
mite is by far the most common (75.5%)
with 56% of them being monosensitized !
Sensitization to olive tree pollen is the sec-
ond most common (32%) followed by
parietaria (30%), cat dander(20%), moulds
(17%) and to grasses(9.5%). We also evalu-
ated 42 food allergic patients .23 of them
were <6 years old and 12 had an IgE-
mediated reaction, 6 to milk, 5 to egg and 1
to peanut. On the other hand,we had 11
babies <6 months who presented with non
IgE mediated reactions(9 allergic proctoco-
litis,2 milk FPIES).In older children,sh is
the main allergen. We also recorded 35 chil-
dren with eczema with only 2 severe cases.
Conclusion: In our selected and small sam-
ple, asthma and rhinitis have a rather high
co-prevalence. The high monosensitisation
rate to dust mite is also remarkable. Olive
tree and parietaria pollen sensitizations are
in accordance with the high concentrations
of these pollens in Crete. Food allergy
seems to be rather rare and mild but rising.
Eczema is also rare.
1476
Adverse drug reactions in a pediatric
allergy unit
Moreno Alvarezz, A1; Vila Sexto, L1; Lopez Calvino, B2;
Pineiro, L3; Moreno Ares, D3
1
A Coruna University Hospital, Pediatric Allergy Unit, A
Coruna, Spain; 2A Coruna University Hospital, Unit of
Clinical Epidemiology and Biostatistics, A Coruna,
Spain; 3A Coruna University Hospital, Pediatric
Department, A Coruna, Spain
Background: Adverse drug reactions
(ADR) are not uncommon in children,
being beta-lactam antibiotics the drugs
implicated most frequently. Few studies
regarding drug allergy (DA) have been per-
formed in pediatric population. We
describe incidence, clinical characteristics
and reliability of diagnostic methods in a
population of children referred to our
Pediatric Allergy Unit between January
2008 and June 2010.
Method: Clinical data from 239 children
(50.6% boys) with suspected DA were
reviewed. Skin tests (prick, intradermal
and patch test) and serum specic IgE
(CAP-FEIA, Sweden) were performed.
Oral challenge (OCs) was performed in
patients with negative skin test, except in
those reporting severe reactions.
Results: 283 allergological work-up were
done. Mean age at the time of ADR was
4.03 years ( 3.42). Mean age at the time
of the allergological work-up: 6.38 years
(3.96)years. Time elapsing between the
onset of symptoms and the date of the study
was 2.34 years (3.21). Thirty-three per-
cent of children suffered from atopic condi-
tions (asthma 59.2%, rhinitis 46.5%, atopic
dermatitis 23.2% and food allergy 4.6%).
They were sensitized to dust mites (86.1%),
pollen (25%) and food (milk, egg and sea-
food: 2.8% each one). Drugs involved were
amoxicillin-clavulanic (52.8%), amoxicillin
(16%), cefuroxime (5.7%), ibuprofen
(5.3%) and cefaclor (3.9%). Fifteen percent
of reactions were immediate and 64.6%
delayed. Clinical manifestations: exanthema
(44.3%), urticaria (26.6%), angioedema
(5.7%), urticaria and angioedema (6.4%)
and serum sickness-like (6%). One patient
presented anaphylaxis. Prick and patch tests
were positive in 1 (0.4%) and 2 (0.7%) of
children, respectively. OCs was positive in
8% of children (29.4% immediate and
70.6% delayed). OCs was positive in 15%
of negative prick test, 5.4% of negative
patch test and 12.9% of negative intrader-
mal test. Overall incidence of drug allergy
was 9.1%. Age of the reaction, sex and per-
sonal history of atopy were not signicant
risk factors for DA.
Conclusion: DA in infants is uncommon.
Most of DA reactions were diagnosed by
OC. Skin test reliability for the diagnosis
of DA in children is limited.
1477
A study on prevalence of positive atopy
patch test in children
Fuiano, N1; Fusilli, S2; Incorvaia, C2
1
ASL Foggia - Distretto di San Severo, Pediatric Allergy
Service, San Severo, Italy; 2Italy
Background: The atopy patch test (APT)
was introduced to assess the cell-mediated
component of the allergic response to envi-
ronmental allergens. A number of studies
attributed to APT increasing importance in
the diagnosis of allergic diseases, showing
that not rarely it may be the only positive
test. We evaluated the prevalence of posi-
tive APT in an unselected population of
subjects in pediatric age.
Methods: The study population was
formed by all children attending a primary
557

and middle school in San Marco in Lamis
(Puglia region, Italy, 12000 inhabitants).
They were 456 children (244 males, 212
females, age ranging from 88 and
167 months, mean age 129.4 months). All
subjects underwent APT and skin prick
tests (SPT) with the most common aller-
gens present in the area (grass, cypress,
Parietaria, and Compositae pollen, Alter-
naria, house dust mites, cat epithelium) by
material from Chemotechnique (Vellinge,
Sweden) for APT and from Stallergenes
(Antony, France) for SPT. A standardized
questionnaire dealing with nose, eye, lungs,
and skin symptoms was administered to
childrens parents.
Results: Among the 456 children, 78
(17.1%) had a positive SPT, and 57
(12.5%) had a positive APT. Sixty-one
subjects (13.4%) were positive only to
SPT, 40 (8.8%) only to APT and 17
(3.7%) to both tests. The allergen most fre-
quently positive was the dust mite (41 SPT
and 55 APT). The peak of positive result
to APT was in the months from January
to May.
Conclusions: This epidemiologic study
shows that a positive APT is quite com-
mon in an unselected pediatric population.
The exclusive positivity to APT in about
9% of children suggests the need to add
APT to common allergy testing, in order
to recognize a cell-mediated sensitization in
subjects otherwise classied as nonallergic.
This seems of particular importance when
dust mites are the causative allergen.
1478
Incidence of sick building syndrome
among pupils in relation to microbial
components in dust in schools in China
Zhang, X1; Zhao, Z2; Larsson, L3; Sebastian, A3;
Norback, D4
1
Shanxi University, Research Center for Environmental
Science and Engineering, Taiyuan, China; 2Fudan
University, Department of Environmental Health,
Shanghai, China; 3University of Lund, Department of
laboratory Medicine, Division of Medial Microbiology,
Lund, Sweden; 4Uppsala University and University
Hospital, Department of Medical Sciences, Uppsala,
Sweden
Background: There are few longitudinal
studies on sick building syndrome (SBS),
which includes ocular, nasal, throat, der-
mal symptoms, headache and fatigue. The
aim was to study associations between new
onset of SBS in Chinese children and aller-
gens and microbial components in the
schools (N = 1993).
Method: A two-year prospective analysis
in a random sample of schools in Taiyuan,
China. Settled dust in the classrooms was
collected by vacuum cleaning and analysed
for lipopolysaccharide (LPS), muramic acid
(MuA) and ergosterol (Erg). Airborne dust
558
Poster Session 62 Pediatric allergy: assessing the burden of disease
was collected in Petri dishes and analysed
for cat and dog allergens and fungal DNA.
The relationship between concentration of
allergens and microbial compounds and
new onset of SBS was analyzed by multi-
level logistic regression.
Result: The prevalence of mucosal and
general symptoms was 33% and 28% at
baseline, and increased during follow-up
(P < 0.001). At baseline, 27% reported at
least one symptom that improved when
away from school (school related symp-
toms). New onset of mucosal symptoms
was negatively associated with concentra-
tion of MuA (P < 0.05), total LPS
(P < 0.001), and shorter lengths of 3-
hydroxy fatty acids from LPS, C14
(P < 0.01), C16 (P < 0.001) and C18
(P < 0.01). Onset of general symptoms
was negatively associated with C18 LPS
(P < 0.01). Onset of school related symp-
toms was negatively associated with C16
LPS (P < 0.05), but positively associated
with total fungal DNA (P <0.01).
Conclusion: Bacterial compounds (LPS and
MuA) seems to be protective for the devel-
opment of mucosal and general symptoms,
but fungal exposure measured as fungal
DNA could be a risk factor for incidence
of school related symptoms.
1479
Immunisation of children with asthma
with H1N1 influenza A vaccine and the
attitudes of the parents
Uysal Soyer, O1; Buyuktiryaki, B2; Erkocoglu, M3;
Dogan, A1; Azkur, D3; Kocabas, C3; Dallar, Y1; Tuncer,
A2; Sekerel, B2
1
Ministry of Health of Turkey, Ankara Education and
Research Hospital, Pediatric Clinic, Turkey; 2Hacettepe
University, School of Medicine, Pediatric Allergy and
Asthma Unit, Ankara, Turkey; 3Ministry of Health of
Turkey, Ankara Hematology and Oncology Education
and Research Hospital, Pediatric Allergy Clinic, Turkey
Background: Pandemic H1N1 inuenza A
virus has caused Swine u worldwide.
Immunization with H1N1 inuenza vaccine
was advised in certain risk groups [chronic
lung disease (asthma, chronic obstructive
pulmonary disease), diabetes mellitus,
chronic renal failure] to prevent the disease
but desired prevalence of vaccination has
not been achieved. Our aim was to evaluate
the prevalence of immunization of children
with asthma with H1N1 Inuenza A vaccine
and the attitudes and opinions of parents
about H1N1 Inuenza A vaccine and the
factors related with acceptance of vaccine.
Method: Between 1 April-30 July 2010, a
survey concerning questions about H1N1
Inuenza A disease and vaccine were
administered to parents of children with
asthma aged 618 years were administered.
Asthma disease related data of the children
were recorded by the physicians.
Allergy 66, Suppl. 94 (2011): 482642  2011 John Wiley & Sons A/S
Result: In the study, 635 patients were
included. During 20092010 inuenza sea-
son, 105 patients (16.8%) were immunized
with H1N1 Inuenza A vaccine and 286
(45.7%) of children with seasonal inuenza
vaccine. The most common reason for
H1N1 Inuenza A vaccine acceptance was
suggestion of the vaccine by the physician.
The reasons for the disclaim of immuniza-
tion were fear of side effects (88.6%), sug-
gestions of politicians (25.2%), media
(24.9%) and friends (18.3%) about rejec-
tion of vaccination. H1N1 Inuenza A vac-
cine acceptance was higher in children with
asthma whose fathers education levels
were high school-university [OR:2.407
(95% CI:1.3244.479), P = 0.004], whom
had school absenteeism due to asthma at
least once in last year [OR: 2.325 (95%
CI:1.0175.311), P = 0.045], whom had
seasonal inuenza vaccination [OR: 2.447
(95% CI:1.3384.476), P = 0.004] and
whose household were also vaccinated
against H1N1 Inuenza A [OR: 12.823
(95% CI:6.637-24.774), P < 0.001]. There
was no relation between H1N1 Inuenza A
vaccination and gender, family history of
asthma and asthma related parameters
(severity, atopy, lung function tests).
Conclusion: The prevalence of H1N1 Inu-
enza A vaccination was lower than
expected in children with asthma. High
sociocultural level, school absenteeism due
to asthma and vaccination with seasonal
inuenza virus affected positively the
H1N1 Inuenza A vaccination acceptance
but environmental interactions (politicians,
media) had a negative impact on vaccina-
tion. To realize the reasons for this unsuc-
cesful vaccination campaign might guide
the future vaccination campaigns.
1480
Safety of rush immunotherapy with de-
pigmented glutaraldehyde-modified dust
mite extracts in children
Costa-Colomer, J1; Bosque, M1; Perea, E2; Gonzalez,
M3; Roca, G3; Valdesoiro, L1; Larramoma, H1; Penas, A4;
Asensio, O1; Garcia, M1
1
Hospital de Sabadell. Universitat Autonoma de
Barcelona, Pediatric Allergy Unit, Sabadell, Spain;
2
Laboratorios Leti, Spain; 3Hospital de Sabadell.
Universitat Autonoma de Barcelona, Pediatric
Department, Sabadell, Spain; 4Hospital Josep Trueta,
Pediatric Allergy Unit, Girona, Spain
Background: Immunotherapy (IT) with
modied allergens reduces allergic asthma
and rhinitis symptoms and medications
requirements. The major risk of allergen
immunotherapy is anaphylaxis. Glutaralde-
hyde-modied therapeutic allergic vaccines
are safer than unmodied vaccines, while
retaining clinical efcacy. A depigmenta-
tion step before the polymerization with
glutaraldehyde has been developed. This
Poster Session 62 Pediatric allergy: assessing the burden of disease
previous step removes non allergic mate-
rial, purify the allergen extract, inactivates
the enzymatic activity of mite extracts and
enhances the solubility of the nal aller-
goid. The aim of this study was to evaluate
the safety of depigmented glutaraldehyde-
modied allergen extracts in a paediatric
group of patients undergoing rush sche-
dule.
Method: One hundred and twenty-three
children between 318 years old, 73 male
and 50 female with a positive history of
IgE mediated allergic rhinitis and/or
asthma due to sensitization against dust
mites allergens received specic subcutane-
ous immunotherapy with depigmented and
polymerized dust mite extracts, adsorbed
onto aluminium hydroxide (Depigoid -La-
boratorios Leti, Spain). The schedule of
administration consisted of a two injections
on the rst treatment day (0.2 + 0.3 ml)
with an interval of 30 min between them
with an observational period of 1 h after
the second dose. This was followed by 1
injection (0.5 ml) monthly. For the evalua-
tion of safety side effects were documented
for each patient.
Result: The safety data of 123 patients
were analyzed. One systemic adverse reac-
tion was recorded (rhinitis) but all patients
completed the schedule treatment. Forty-
two immediate local reactions appeared, 26
of them <5 mm and 16 >5 mm.
Conclusion: Results of this study suggested
evidence that this schedule IT with depig-
mented gluteraldehyde-modied allergen
extracts was well-tolerated.
1481
Allergic diseases among the children and
adolescents in the CIS-countries
Slavyanskaya, T
Russian University, Allergology and Immunology,
Moscow, Russian Federation
Background: To summarize the results of
the CIS countries (CIS-C)research on the
prevalence, structure and peculiarities of
the course of allergic diseases(AD) among
children(Ch) and adolescent(Adl) for the
last 5-10 years.
Materials: The results of the research of
the ISAAC and ARIA programs, obtained
from the National Societies of Allergology
and Immunology, statistic reports of the
Republic Ministries of Health and medical
institutions on the dissemination of AD in
children, living in industrial cities and rural
areas.
Results: The analysis of the data has
showed that the structure of AD CIS-C is
dominated by: bronchial asthma (BA),
allergic rhinitis(AR), atopic dermati-
tis(AD), pollinosis(P) and allergic conjunc-
Allergy 66, Suppl. 94 (2011): 482642  2011 John Wiley & Sons A/S
tivitis(AC). AR, on average, has been
diagnosed in 10.5% (from 2 to 34.7%)
Ch+Adl. AP, according to the negotiabil-
ity has hurt only 0.10.4% of the popula-
tion. BA in the CIS-C, on average, suffers
431% of child population. The highest
incidence of BA has been observed in
Armenia, Belarus, Moldova, Ukraine, Ky-
rgyzstan, Russia, Tajikistan. The actual
incidence among Ch+Adl was 2140%. In
this case, the diagnosis of BA in the insti-
tutions of practical public health has been
recorded in Adl-3.2% and in Ch-1.5%.
The average incidence of AD in the CIS-C
in Ch+Adl is 2.220.3%. The high preva-
lence of blood pressure is characteristic of
Ukraine, Tajikistan, Kazakhstan, Kyrgyz-
stan, Belarus. In Armenia and Tajikistan
the level of AD diseases among Ch+Adl
has been noted within 2.74.7%. In Tajiki-
stan, Belarus, Kazakhstan and Armenia
prevalence of P among child population is
4.35.2%. Among all CIS-C AC most
often has been met in Armenia, Tajikistan,
Kazakhstan, Azerbaijan, Georgia. In
Armenia a signicant incidence of AC is
established among Ch+Adl (39.162.2%).
The prevalence of AD depends on urbani-
zation in the region and the climate and
geographic conditions. In the highlands
AD are extremely rare. The prevalence of
AD is 23 times higher in industrial areas.
Conclusion: Studies have shown that
among the child population the patients
with mild disease were dominating. The
structure of AD recorded by uptake, domi-
nated the disease with medium-severe
course. In all CIS-N there have been seen
the hypodiagnostics of all forms of AD.
Accounting for the structure and expres-
sion of sensitization to allergens, taking
into account climate and geographic fea-
tures, allows to plan the work of pediatric
allergic services and to organize preventive
and curative interventions.
1482
Evaluation of the patients with recurrent
croup
Usta Guc, B1; Asilsoy, S1; Ozer, F2; Durmaz, C1
1
Baskent University Hospital, Pediatric Allergy, Turkey;
2
Baskent University Hospital, Otolaryngology Head and
Neck Surgery, Turkey
Background: Croup is a disease frequently
seen in childhood. A lot of children consult
to Child Emergency Service and outpatient
clinics for this reason.
Material and Methods: In this study
patients consulting to outpatient clinic due
to recurrent croup (RC) were retrospec-
tively evaluated. Patients medical histories,
ndings of the physical examination and
investigations conducted were reviewed.
Results: 51 patients, 42 of whom were
males and whose ages were
4.69 3.23 years (5 month14 years) were
evaluated. It was found out that wheezing
accompanied with the croup complaints in
37 of the patients (72.5%), it was sudden
onset in 19 patients (37%), it triggered by
viral infections in 15 patients (29.4%), it
was sometimes sudden onset and some-
times induced viral infections in 17 patients
(33.3%). Gastro-oesophageal reux was
examined in 31 patients, and 13 of them
was detected positive (25.5%). Laryngotra-
cheobronchitis in 30 of the patients, atopic
dermatitis in 9 of them, allergic rhinitis in
26, and family history of atopy in 25 of
them were detected. Skin prick tests were
performed in 48 of the patients (94%), and
atopy was established in 21 of them
(41.2%). Findings displayed treatment-
resistant characteristics in 16 patients.
Direct laryngoscopy was conducted in 10
of the patients, and anomaly was identied
in 6 patients. Three of the patients with
RC were under the age of 1; in these
patients reux examination was carried out
as barium swallow for a probable vascular
ring and anatomical anomaly, and appar-
ent reux was detected in 2 patients.
Conclusion: Recurrent croup is a clinical
concept. Even though it is frequent in pedi-
atric population, there are not denitions
concerning its etiology, natural course and
epidemiology. It is signicant to examine
the patients in terms of the underlying
pathologies and other contributing factors.
1483
Urticaria to clothes containing cobalt
pigment
Pauperio, S; Mendonca, V; Bonito Vitor, A
Hospital de S. Joao do Porto, Pediatric Allergy
Department, Porto, Portugal
Background: Allergy to cobalt is one of the
most common specially in female adoles-
cents and is easily treatable when diag-
nosed. Contact with metals (nickel,
chromium and cobalt) increases exponen-
tially during school age and adolescence,
taking responsibility for the majority of
contact dermatitis occurring in this period.
Allergic contact dermatitis (ACD) is a
common clinical entity in children and
adolescents, with an estimated prevalence
between 13.3 and 24.5%, increasing with
age. Occurs in individuals with atopic pre-
disposition and involves an immune mech-
anism mediated by T lymphocytes occurs
in two phases: the sensitization phase and
elicitation phase. Numerous substances
have been described as causative factors
depending on the age at onset, socio-eco-
nomic development and new contact with
559

haptens. Prevalence of ACD is increasing
in childhood and adolescence mainly
attributed to the increased chemical expo-
sure in this age group, timely recognition
by pediatricians and patch testing.
Method: A 14-year-old female followed in
Pediatric Allergology by rosaceae allergy
related with non specic lipid transport
proteins (Ns LPTs) diagnosed a year ago.
She referred a reaction that occurs in a
class starting immediately with an itchy
rash of upper limbs and face associated
with dyspnea. She denied food intake of
the Rosaceae family, but associated with
the use of a blue vest in the class. Some
months before, a similar episode occurred
when she put a colored sweater, dark blue
(for ten minutes), with the onset of an urti-
cariform rash of the trunk, upper limbs
and face with the sensation of dyspnea
which motivated the recourse to emergency
department. Performed skin tests of the
standard series, patch testing, textile dyes
and sweatshirt, which read at 48 h was
positive for cobalt. In addition to rosaceae
eviction was also instituted eviction contact
with ornaments, blue dyes or paints con-
taining cobalt such as clothing, glass, cera-
mic, cosmetics (antitranspirants, shadows
and hair dyes) and B12 vitamin. She has
an adrenaline pen for self-administration
prescribed for food allergy reactions.
Conclusion: The authors present this case
for its singularity, because although the
allergic contact dermatitis to cobalt is rela-
tively common, the earliness and severity
of symptoms reported by this adolescent
suggests an IgE-mediated reaction.
560
Poster Session 62 Pediatric allergy: assessing the burden of disease
1484
Horse allergy in atopic children: a clinical
lesson
Stadermann, M1; Meijer, Y1; Pasmans, S2; Ent van der,
K3
1
Wilhelmina Childrens Hospital, Pediatric Pulmonology
and Allergology, Utrecht, Netherlands; 2Wilhelmina
Childrens Hospital, Pediatric Allergology, Utrecht,
Netherlands; 3Wilhelmina Childrens Hospital, Pediatric
Pulmonology, Utrecht, Netherlands
Background: Data on horse allergy in chil-
dren are scarce. Hence the predictive value
of positive IgE for horse to estimate the
risk of a possible allergy for horses in the
individual patients is unknown. From our
own experience we know that children who
are sensitized for horse can become ana-
phylactic when exposed to horses. This
study was conducted to investigate the pre-
dictive value of IgE levels for horse allergy.
Method: From our outpatient clinic we
selected all children who were tested for
specic IgE for horse between July 2007
and December 2010. All patients had clini-
cal suspicion on horse allergy. We retro-
spectively reviewed the patients clinical
history and assessed the severity of allergic
symptoms.
Result: Seventy-ve children (44 male) with
mean age of 4.6 (2.38) years were identi-
ed. One patient with Nethertons syn-
drome was excluded. The other children
were clinically atopic with eczema (67/74),
asthma (58/74), food allergy (66/74) and/or
allergic rhinitis (59/74). As far as allergic
symptoms to horses were concerned, four
different categories could be distinguished.
First children without clinical symptoms
(n=4). Median IgE levels of these children
Allergy 66, Suppl. 94 (2011): 482642  2011 John Wiley & Sons A/S
were 1.5 (range 0.43.9) kU/l. Secondly,
children who developed urticaria, angio-
edema and nasal discharge after exposure
to horse (n = 20). Median IgE level in this
group was 8.4 (range 0.7968) kU/l.
Thirdly, children who in addition to urti-
caria, angio-edema and nasal discharge
reported instant wheezing, stridor, dysp-
noea or even collapse (n = 43). Median
IgE levels in this group were 21.5 (range
0.7>100) kU/l. In seven children the
prevalence of horse allergy was unclear as
contact with horses was strictly avoided or
medical data describing allergic reaction
after exposure to horse were incomplete.
All allergic symptoms occurred within 1 h
after exposure to a horse. Parents reported
stress and anxiety during the allergic reac-
tion. None of the children received treat-
ment other than anti-histaminica and
sympathicomimetic drugs. Four children
required emergency treatment at a hospi-
tal.
Conclusion: Serum IgE level for horse is
associated with clinical suspicion to horse
allergy. Horse allergy can result in severe
urticaria and respiratory symptoms within
one hour after exposure. In case of sensi-
bilisation for horse, measures should be
taken to avoid contact with horses and to
train parents how to adequately handle
allergic reactions.

Pediatric asthma: clinical assessment and therapy
1485
Broncho-obstructive syndrome in child-
hood - is it always asthma?
Muti, A1; Iacob, D2; Farcau, D2; Fufezan, O3
1
UMF, Immunology and Allergy, Cluj-Napoca, Romania;
2
UMF Iuliu Hatieganu, Paediatrics, Cluj-Napoca,
Romania; 33rd Paediatric Clinic, Radiology, Cluj-
Napoca, Romania
Clinical presentation of the broncho-
obstruction in children is most often highly
suggestive of bronchiolitis, recurrent
wheezing or asthma. We present the cases
of two patients with recurrent bronchiolitis
and asthma, non-responsive to treatment.
The rst female patient presented 2 epi-
sodes of uncomplicated bronchiolitis dur-
ing the 7th month of life and a 3rd episode
of bronchiolitis complicated with pneumo-
nia during the 8th month of life. When
admitted for the 3rd episode, she presented
an oxygen saturation of 91% in ambient
air. Thoracic ultrasounds oriented the diag-
nosis towards a diaphragmatic hernia, con-
rmed through barium swallow and
barium enema. The surgical treatment of
the hernia determined the resolution of
respiratory symptoms.
In the second case, a 9-year-old boy pre-
sented a broncho-obstructive syndrome
interpreted as allergic asthma associated to
allergic rhinitis with sensitization to house-
dust mites and dog. A treatment with
inhaled corticosteroids and antihistamine
was prescribed with little improvement of
the asthma symptoms. Six months later the
patient presented for vomiting and produc-
tive cough. Thoracic ultrasound suggested
achalasia, diagnosis conrmed through
esophageal manometry and barium swal-
low. Surgical treatment led to resolution of
asthma-like symptoms with persistence of a
mild intermittent rhinitis.
Unfavourable clinical course, despite
correct treatment in both cases required
additional investigations which nally led
to the correct diagnosis and treatment.
Conclusion: For the differential diagnosis
of non-responding broncho-obstructive
syndrome in children, one must think to
digestive diseases. Ultrasound was the elec-
tive non-invasive method in diagnosing
these cases.
Allergy 66, Suppl. 94 (2011): 482642  2011 John Wiley & Sons A/S
Poster Session 63
1486
The incidence of cystic fibrosis in pediat-
ric patients evaluated as asthma
Hatipoglu, H1; Erkal, S2; Hatipoglu, N3; Turel, O3;
Aydogmus, C3; Babayigit, A3; Engerek, N3; Turkmen, S4;
Kurt, K5; Aldemir, H3; Yesil, Y3; Siraneci, R3 Gastroesophageal reflux disease is it a
1
Istanbul Bakirkoy Maternity and Childrens Hospital,
Pediatric Allergy Immunology and Infectious Diseases
Dept., Istanbul, Turkey; 2Istanbul Bakirkoy Maternity
and Childrens Hospital, Biochemistry Dept., Istanbul,
Turkey; 3Istanbul Bakirkoy Maternity and Childrens
Hospital, Pediatric Allergy Immunology and Infectious
Diseases Dept., Turkey; 4Istanbul Bakirkoy Maternity
and Childrens Hospital, Biochemistry Dept., Turkey;
5
Istanbul Bakirkoy Maternity and Childrens Hospital,
Turkey
Background: Cystic brosis is a chronic
and genetically transmitted disease affect-
ing respiratory, gastrointestinal, and repro-
ductive systems as well as pancreas and
biliary tract. This morbidity must be con-
sidered in patients with recurrent respira-
tory symptoms and asthma-like signs. The
chlorine level above 60 mmol/l in sweat
has been accepted as diagnostic for cystic
brosis in patients with compatible clinical
ndings. The denitive diagnosis can be
made after A second measure of chlorine
level in sweat at a different laboratory.
Method: This study was performed
between September 2009 and August 2010.
Children aged 018 years old with at least
three episodes of acute wheezing were
enrolled in this study. Sweat was collected
and chlorine level was measured in every
patient besides routine work-up tests for
recurrent wheezing (tuberculin skin test,
chest X-ray, immune-globulin levels). The
sweat test was performed in allergy labora-
tory of our hospital by Wescor Swert-
Check Conductive Osmometer analyser
and results were recorded.
Results: One hundred and eighty patients
have been admitted to our allergy outpa-
tient clinic with chronic asthma during last
year and sweat tests were positive in two
of them. The test was repeated for each
patients and found to be positive in
another laboratory, conrming the diagno-
sis of cystic brosis.
Conclusion: Early diagnosis is essential for
cystic brosis. The relatively short life span
could be improved with adequate nutrition,
combating with infections, respiratory
physiotherapy, but timely diagnosis is of
utmost importance. Sweat chlorine test
should be used as a routine and noninva-
sive study for pediatric patients with recur-
rent respiratory symptoms.
1487
frequent comorbidity in pediatric
asthma?
Hatipoglu, H1; Hatipoglu, N1; Aydogmus, C1; Turel, O1;
Babayigit, A1; Engerek, N1; Kurt, K1; Aldemir, H1; Yesil,
Y1; Erkal, S2; Turkmen, S2; Siraneci, R1
1
Istanbul Bakirkoy Maternity and Childrens Hospital,
Pediatrics Dept., Istanbul, Turkey; 2Istanbul Bakirkoy
Maternity and Childrens Hospital, Biochemistry Dept.,
Istanbul, Turkey
Background: Gastroesophageal reux dis-
ease (GERD) is frequently found in associ-
ation with asthma. Successful control of
GERD in these patients may improve in
their asthma symptoms. The diagnosis is
established by scintigraphic investigation
or 24-hour pH monitoring or even esopha-
geal and gastric endoscopy. The latter two
methods are invasive, as both methods
need hospitalization, patient compliance
and skilled personnel. However, scintigra-
phy is more practical and widely reached.
Method: The coexistence of GERD in
asthmatic children has been studied using
scintigraphic method. This study was per-
formed between September 2007 and
August 2009. Patients aged 6 months and
14 years old, suffering from at least three
episodes of acute wheezing, and/or recur-
rent nausea and vomiting were included.
Radionuclide scintigraphy was performed
and any reux from gastric content to
esophagus was monitored in every patient,
who also underwent routine work-up tests
for recurrent wheezing (sweat and tubercu-
lin skin tests, chest X-ray, immune-globulin
levels).
Result: A total of 100 patients were
enrolled. Forty cases had positive result on
radionuclide scanning. There were 43 chil-
dren aged between 6 months2 years and
20 (46.5%) of them had GERD according
to test results, whereas 20 patients in the
group older than 2 years (35.1, total 57
patients) had reux.
Conclusion: GERD was found relatively
frequently in pediatric patients less than
two years of age. Effective control of
asthma can be achieved successfully if a
comorbid GERD will be treated properly.
Esophageal scintigraphy, because of its
561
 Poster Session 63 Pediatric asthma: clinical assessment and therapy

perfect tolerance and reliability, seems to asthma severity, atopy, thickness of atelec- Conclusion: The children with SCD show
be an examination that can be referred to tasis at admission, etc. more frequent allergy and asthmatic symp-
such a situation that is easily reached and Conclusion: Early diagnosis and treatment toms in comparison with the controls simi-
applied. of right middle lobe syndrome prevent larly to other studies performed in other
complications. The patients with inade- populations. The complexity of the disease,
quate response to treatment in the rst the importance of the therapy with
5 days might recover in long term and hydroxyurea, the difculty to nd normal
1488
should be followed closely for development controls in families with a high prevalence
Right middle lobe syndrome in children
with asthma and prognostic factors
of complications. of heterozygous subjects are important
confounding factor in these studies. How-
Uysal Soyer, O1; Ozen, C1; Senyucel, C2; Dallar, Y1 ever these preliminary data could underline
1
The Ministry of Health of Turkey, Ankara Education
and Research Hospital, Pediatric Clinic, Ankara, Turkey; the signicant overlap in the pathophysiol-
1489
2
Kirikkale University, School of Medicine, Radiology ogy of asthma and pulmonary insults in
Department, Kirikkale, Turkey
Allergy and asthma in children with
sickle cell disease coming from Africa: a
SCD. A correct diagnosis of allergy and
preliminary study asthma and a correct asthma therapy may
Background: Segmental or lobar collapse be important in delaying or averting the
of lungs is a complication of asthma and Perino, A; Lamon, M; Serra, M; Roggero, S
progression of sickle cell lung disease.
right middle lobe is most commonly University of Torino, Pediatrics Department, Torino,
Italy
involved. The aim of this study is to dene
the characteristics and prognostic factors
Background: Sickle cell disease is a genetic 1490
of right middle lobe in asthma during
disorder of haemoglobin transmitted on an Dysfunctional voiding in children with
childhood.
autosomal recessive basis. Homozygosity asthma
Method: In this prospective study, children
for the HbS allele (haemoglobin SS dis-
with asthma diagnosed as right middle lobe Uysal Soyer, O1; Kilincoglu, B1; Fedakar Senyucel, M2;
ease) is the most common nding. Hetro- Arslan, K2; Cakmak, M2; Dallar, Y1; Soyer, T2
atelectasis between April 2009 and Septem-
zygous subjects are healthy. Pulmonary 1
The Ministry of Health of Turkey, Ankara Education
ber 2010 were included; demographic char-
complications account for signicant mor- and Research Hospital, Pediatric Clinic, Ankara, Turkey;
acteristics and thickness of atelectasis at 2
Kirikkale University, School of Medicine, Pediatric
bidity and mortality in the SCD patients.
admission were noted. Systemic methyl- Surgery Department, Kirikkale, Turkey
Asthma is a common comorbidity and a
prednisolone (2 mg/kg/day, 5 days), claritr-
risk factor in sickle cell disease (SCD) with Background: Dysfunctional voiding (DV)
omycine (25 mg/kg/day, 10 days), inhaled
a reported prevalence of 3070%.The most is a common symptom during childhood.
salbutamol were given and chest physio-
studies are performed in Afroamerican Children with asthma might have increased
therapy (onset on sixth day) were applied.
patients who are largely affected by intra-abdominal pressure due to chronic
Patients were reevaluated on day 6, 14, 30
asthma. cough and dyspnea. The relationship
and 90 days and the chest X-rays were
Method: In order to determine the preva- between increased intraabdominal pressure
taken if the atelectasis did not resolve in
lence of asthma and allergy in young and DV was well documented. Our aim
the previous visit.
patients with SCD coming from Africa, was to evaluate DV in children with
Result: Twenty-nine patients (6.8 2.13
with a different genetic and environmental asthma and to compare with healthy sub-
years, male 50%) were enrolled in the
background in comparison with population jects.
study. Atopy was determined in 12
studied in US, we studied 43 children with Method: Children aged between 410-year-
(44.4%) patients. In the last year, patients
SCD (HbS/HbS) and some hemoglobinic old with asthma (n = 178) and healthy
had experienced frequent asthma attacks
variants (HbS/Thal;HbS/HbC;HbS/Oa- subjects (n = 197) were enrolled in the
[n = 3 (25.75)] and 19 children were hos-
rab;HbS/HPHF) and 10 normal controls study. Validated Dysfunctional Voiding
pitalized at least once due to asthma.
matched by age and race. We performed and Incontinence Scoring System (DVISS)
Onset of symptoms was 20 (710) days
skin prick test and/or specic IgE dosage were administered to all subjects and
before admission. The most common pre-
for a large panel of respiratory allergens, demographic features and disease related
senting symptoms were cough (100%), spu-
respiratory function test and FENO in all characteristics in children with asthma
tum production (85.7%) and dyspnea
the children more than 3 years old fol- were recorded. Children with DVISS score
(71.4%). At admission, the thickness of
lowed in our department for SCD, both greater than 8.5 were diagnosed as sus-
atelectasis was 12.08 1.11 mm; FEV1%
complaining respiratory symptoms or not. pected DV.
were 74.9 14.21 and Childhood Asthma
Result: 14 patients out of 43 (32.5%) had Result: There was no difference between
Control Test scores (C-ACT) were
positive skin prick tests and 2 controls age and gender of children with asthma
12.2 5.88. The atelectasis were resolved
(20%). 7 patients with SCD had obstruc- (6.85 1.7 years, 53.3% male) and
on sith day (n = 3), on fourteenth day
tive pulmonary disorders (16%) and 2 healthy subjects (6.75 1.57 years, 58.9%
(n = 9), on thirtieth day (n = 10) and on
showed pulmonary restriction. Pulmonary male) (P > 0.05). Sixty-ve of asthmatic
ninetieth day (n = 3). The treatment
test were normal in the controls. FENO patients were younger than 6 years of age
response of the patients whose atelectasis
could be not assessed in many small chil- and 110 of them were older than 6 years of
resolved in 14 days were better on sixth
dren for lacking compliance. Furthermore age. In children younger than 6 years of
day [atelectasis thickness: 4.68 1.71 and
we are trying to asses the reference values age, DV symptoms were noted in 19
11.94 2.0 mm, P = 0.020] compared to
of FENO for this type of patients who (29.2%) of asthmatic patients and 5 of
the patients whose atelectasis resolved
could have some basic disturbances in (6.8%) of healthy subjects (P < 0.001).
later. Also, C-ACT scores of the patients
metabolism of NO due to the disease. Only When factors effecting dysfunctional void-
whose atelectasis resolved in 14 days were
the 5% of the patients studied complained ing were evaluated in children younger
higher on thirtieth day [22 (21.524) and
for asthma symptoms. Many of them than 6 years of age, it revealed that asthma
17 (1421), P = 0.008]. Regression analy-
showed adenotonsillar hypertrophy as con- had increased DV symptoms 5.7 times
sis did not reveal any prognostic factor
founding factor. (95% CI=1.98816.344) than healthy con-
affecting recovery of atelectasis including

562 Allergy 66, Suppl. 94 (2011): 482642  2011 John Wiley & Sons A/S
Poster Session 63 Pediatric asthma: clinical assessment and therapy
trols. Children with asthma older than
6 years of age had similar prevalence of
DV and nocturnal enuresis symptoms
(P > 0.05) compared to healthy children
but they had higher frequency of voiding
(>7 times/day) [29 (26.4%) and 17
(13.9%), P = 0.018] and had higher preva-
lence of urgency [36 (32.7%) and 25
(20.5%), P = 0.035].
Conclusion: Presence of asthma might
cause DV symptoms in children who did
not completed their toilet training but
might cause frequency and urgency in
older children. Symptomatic DV should be
kept in mind in children with asthma.
1491
Adolescent and asthma: can physiother-
apy help?
Coelho, C; Nogueira, K
University of State of Rio de Janeiro, Center of Study
of Adolescent Health, Rio de Janeiro, Brazil
Background: The aim of physiotherapy is
to improve the physical tness of adoles-
cents with asthma, providing a gradual
readjustment to physical efforts and
encouraging it to participate in physical
activities in general.
Method: The study was conducted at the
NESA Center of Study of Adolescent
Health from July 2009 to May 2010. Were
studied 6 cases of teenagers, all male, aged
12 to 14 years. The teenagers were initially
evaluated by allergy sector noted that the
need to improve the posture of the same
because only then would result in improve-
ment in mechanical ventilation and help
with treatment for asthma.
Result: Physiotherapy used relaxation tech-
niques, stretching and strengthening the
muscles of the body. Also, promoted the
rehabilitation and functional diaphrag-
matic breathing with awareness of mechan-
ical ventilation. The method of global
postural reeducation (GPR) was used in
conjunction with physiotherapy, aimed at
restructuring posture and breathing the
same time. As sessions were individual and
group intermittently, on average 2 times a
large part semana. The presence of one
member family for guidance and correction
of such exercises should be practiced and
even awareness of the importance of being
practiced daily at home.
Conclusion: In conclusion, improved pos-
ture and stretching shortened muscles, the
accessory muscle relaxation our patients
came to control their disease better, more
spaced crises and improved their quality of
life. Thus conrming the importance of the
multidisciplinary team in the monitoring of
chronic diseases.
Allergy 66, Suppl. 94 (2011): 482642  2011 John Wiley & Sons A/S
1492
the effect of long-term asthma treatment
Clinical predictors for hospital admission with inhaled corticosteroids (ICS) or mont-
due to exacerbation in children with elukast on AH.
asthma Materials and methods: 528 unselected,
maturely born infants were followed-up
Gheonea, C1; Popescu, F2; Florincescu Gheorghe, A3;
Dumitrescu, L4
until they were seven years old. Otolaryng-
1
University of Medicine and Pharmacy of Craiova, ologists diagnosis of AH was based on
Department of Paediatrics, Craiva, Romania; 2University sleep disorders, clinical manifestations
of Medicine and Pharmacy Carol Davila, Department
(nasal congestion, mouth breathing, rhin-
of Allergology, Bucharest, Romania; 3Municipal
Hospital Filantropia Craiova, Allergology, Craiova, orrhea, cough and hyponasal speech) and
Romania; 4University of Medicine and Pharmacy of examination. Diagnosis of allergic asthma
Craiova, Department of Paediatrics, Craiova, Romania
was ascertained with the ISAAC core, spe-
cic IgE testing and positive skin prick tes-
Objective: To identify clinical predictive
tes. At the age of 7 years data of 84
factors of hospital admission due to
children with asthma on ICS or monteluk-
asthma exacerbation in children with
ast controller therapy for at least one year-
asthma attending a Regional Centre for
group A and of 444 healthy children-group
Paediatric Asthma.
B were analysed. In the group A the effect
Method: Retrospective, descriptive study.
of asthma treatment with ICS (subgroup
A database was created including demo-
A1) or with montelukast (subgroup A2) on
graphic and clinical characteristics for a
AH was analysed. Statistical analysis was
period of 12 months of the subjects
performed using chi-square analysis with
(n = 216, age = 9.2 5.8 years, 102
Yates correction. P-values less than 0.01
boys). Descriptive statistics were followed
were considered signicant.
by chi-square tests to identify variables
Results: In the study group 26.1% of chil-
associated with hospitalizations. Logistic
dren had positive history of parental
regression analysis was used to determine
allergy and 17.1% of children had AH.
which variables were signicant predictors
The prevalence of AH was signicantly
of asthma-related hospitalizations.
higher in children with positive (37.7%)
Result: Thirty-two patients experienced a
than in those with negative history of
total of 67 hospitalizations. The only vari-
parental allergy (8.9%), (P < 0.01). As
ables found to be a predictive factor were
well it was higher in children with asthma
greater asthma severity (OR = 19.5; 95%
(26.2%) than in healthy children (15.3%),
CI: 3.8468.57), and previous hospitaliza-
(P = 0.02). Prevalence of AH tended to be
tion (OR = 12.2; 95% CI: 2.7152.1), and
lower in children with asthma treated with
allergic rhinitis (OR = 9.4; 95% CI: 1.04
ICS (subgroup A1: 25.4%) than in those
42.28).
treated with montelukast (subgroup A2:
Conclusion: Our data support the impor-
45%), (P = 0.05). The percentage of chil-
tance of targeting early intervention in
dren with AH who needed adenoidectomy
asthmatic children with greater disease
was a bit lower in the group A (22.7%)
severity and associated allergic rhinitis.
then in the group B (33.8%), (P = 0.47).
Furthermore, results suggest involving chil-
Conclusions: We demonstrated an associa-
dren in individual self-care of their asthma
tion between allergic asthma and AH in a
might improve the clinical course.
group of unselected Slovene children. No
benecial effect of long-term asthma treat-
ment with ICS or montelukast on decrease
1493 of AH prevalence and avoidance of ade-
Does asthma treatment prevents adenoi- noidectomy was conrmed.
dal hypertrophy?
Besednjak-Kocijancic, L
Zdravstveni dom Nova Gorica, Primary Pediatric Health
1494
Centre, Nova Gorica, Slovenia
Efficacy of nebulised budesonide in
infants with viral wheezing
Background: Adenoidal hypertrophy (AH)
is common disorder in paediatric popula- Durmaz, C; Asilsoy, S; Usta Guc, B
Baskent University School of Medicine Faculty,
tion. The causes of AH have not been fully
Pediatric Allergy, Adana, Turkey
elucidated. The role of allergy remains
unexplained. AH might be the consequence
Background: Recent international guide-
of increased production of specic IgE
lines recommend the use of inhaled corti-
within the lymphatic tissue of Waldeyers
costeroids as the controller therapy for the
ring in an allergic child. Recently, has been
management of persistant asthma in young
demonstrated positive role of intranasal
children. The treatment of those children
corticosteroids and montelukast in man-
with intermittent, viral induced wheezing
agement of patients with AH. The aim of
remains controversial. We examined the
this study was to asses an association
efcacy of nebulized budesonide in young
between AH and asthma and to evaluate
563

children with intermittent, viral induced
wheezing.
Method: We randomly assigned 66 chil-
dren who were 636 months age with inter-
mittent, viral induced wheezing to receive
500 mcg/day of budesonide plus salbuta-
mol when needed for three months or
receive only salbutamol when needed. Pri-
mer outcomes were emergency visits and
use of systemic steroid. Secondary out-
comes were any wheezing during upper
respiratory infection, use of salbutamol
and duration of wheezing episodes.
Result: Of the 66 children, 36 received neb-
ulized budesonide and salbutamol when
needed (group 1) and 30 received only neb-
ulized salbutamol when needed (group 2).
The two groups did not differ in age, gen-
der, age of rst wheezing episode, atopic
feature, serum IgE levels and total eosino-
phil counts. At the end of three months no
signicant difference was found in emer-
gency visits, use of systemic steroid, wheez-
ing during upper respiratory infection and
use of salbutamol. But the duration of
wheezing was shorter during upper respira-
tory tract infection in group 1.
Conclusion: Although nebulized budeso-
nide had shorten the duration of wheezing
in children with intermittent, viral induced
wheezing, it had no major effect on emer-
gency visits, use of systemic steroid, wheez-
ing during upper respiratory infection and
use of salbutamol.
1495
Age relationship of asthma exacerbations
in two classic perennial phenotypes, non
atopic and mite monosensitised patients
Petalas, K; Vourdas, D; Grigoreas, C
251 General Air Force Hospital, Allergy, Athens, Greece
Background: Asthma is characterized by
exacerbations and the major morbidity,
mortality, health care costs are strongly
associated with them. Asthma exacerba-
tions (AE) are caused for a variety of rea-
sons including unrecognized disease,
undertreated or unresponsive to conven-
tional therapy asthma, recent exposure to
triggers like viruses, usually rhinoviruses or
allergens. Frequent AE are a feature of
severe persistent asthma but severe exacer-
bations can occur even in patients with mild
or well controlled asthma. AE is dened as
a worsening of asthma sufcient to require
medical intervention and usually adminis-
tration of oral steroids reected objectively
as a fall of PEF, FEV1 20% or 30%. The
ideal way to reduce exacerbations is to pre-
vent them by prophylactic therapy. It is
important though to recognize AE epidemi-
ologic characteristics, like the age of
patients, depending on asthma phenotype.
564
Poster Session 63 Pediatric asthma: clinical assessment and therapy
Table 1. For abstract 1495.
Age 014 1529 3044 >44 Total
Number of Patients 107 146 69 20 342
Number of AE 193 231 116 50 590
Expected Number of AE 184.59 251.87 119.04 34.50
Table 2. For abstract 1495.
Age 014 1529 3044 >44 Total
Number of Patients 690 254 267 307 1518
Number of AE 1329 332 433 491 2585
Expected Number of AE 1175.00 432.54 454.67 522.79
Method: We have studied, in our depart- undertreated or unresponsive to conven-
ment, during 20 years (from 1989 to 2009), tional therapy asthma, recent exposure to
AE in patients suffering from two differ- triggers like viruses, usually rhinovirus or
ent, but with common characteristics (i.e. allergens. Frequent AE are a feature of
perennial), phenotypes of asthma: non ato- severe persistent asthma but severe exacer-
pic and mites monosensitized patients. The bations can occur even in patients with
data in each group was classied into four mild or well controlled asthma. AE is
age categories 014, 1529, 3044 and dened as a worsening of asthma sufcient
greater than 44. The results were evaluated to require medical intervention and usually
by applying the 2 test and the null administration of oral steroids reected
hypothesis. The null hypothesis is that objectively as a fall of PEF, FEV1 >20%
there is not difference in the number of AE or 30%. The ideal way to reduce exacerba-
occurred in the four categories above. tions is to prevent them by prophylactic
Result: Mite monosensitized patients. therapy. It is important though to recog-
By applying the 2-test for 3 degrees of nize AE epidemiologic characteristics, like
freedom (d.f) we obtain 2 = 9.151 which gender of patient, depending on asthma
corresponds to the P = 0.0273. Therefore phenotype.
the null hypothesis is not accepted and the Method: We have studied, in our depart-
difference of the number of AE in the age ment, during 20 years (from 1989 to 2009),
categories is statistically signicant. Non AE in patients suffering from two differ-
atopic patients. ent, but with common characteristics (i.e.
By applying the 2-test for 3 d.f we perennial), phenotypes of asthma: non ato-
obtain 2=46.518 which corresponds to a pic and mites monosensitized patients. The
P < 0.0001. Therefore the null hypothesis data in each group was classied into
is not accepted and the difference of the males and females. The results were evalu-
number of paroxysms in the age categories ated by applying the 2 test and the null
is statistically highly signicant. hypothesis. The null hypothesis is that
Conclusion: Our results indicate that in there is not difference in the number of AE
Greece during 20 years (from 1989 to occurred in the four categories above.
2009) AE are age related in non atopic and
mites monosensitized asthmatics meaning Table 1. For abstract 1496.
that their frequency is higher in younger
Gender Male Female Total
patients.
Number of Patients 214 128 342
Number of AE 370 220 590
Expected Number of AE 369.18 220.82
1496
Gender relationship of asthma exacerba-
tions in two classic perennial pheno- Table 2. For abstract 1496.
types, non atopic and mite
Gender Male Female Total
monosensitised patients
Number of Patients 736 782 1518
Petalas, K; Vourdas, D; Grigoreas, C
Number of AE 1335 1250 2585
251 General Air Force Hospital, Allergy, Athens, Greece
Expected Number of AE 1253.3 1331.7
Background: Asthma is characterized by
exacerbations and the major morbidity, Result: Mites monosensitized patients.
mortality, health care costs are strongly By applying the 2-test for 1 d.f we
associated with them. Asthma exacerba- obtain 2 = 0.005 which corresponds to
tions (AE) are caused for a variety of rea- the P = 0.944. Therefore the null hypo-
sons including unrecognized disease, thesis is accepted and there is not any
Allergy 66, Suppl. 94 (2011): 482642  2011 John Wiley & Sons A/S
Poster Session 63 Pediatric asthma: clinical assessment and therapy
statistical signicant difference in the num-
ber of paroxysms occurred regarding gen-
der. Non atopic patients.
By applying the 2-test for 1 d.f we
obtain 2 = 10.330 which corresponds to
the P = 0.0013. Therefore the null hypoth-
esis is not accepted and there is statistical
signicant difference in the number of par-
oxysms occurred regarding gender.
Conclusion: Our results indicate that in
Greece during 20 years (from 1989 to
2009) AE are gender related in non atopic
asthmatics meaning that their frequency is
higher in males. AE in mites monosensi-
tized asthmatics is not gender related.
1497
Comparison of asthma exacerbations epi-
demics in house dust mite monosensi-
tized and non atopic patients
Petalas, K; Vourdas, D; Grigoreas, C
251 General Air Force Hospital, Allergy, Athens, Greece
Background: Asthma exacerbations (AE)
are caused for a variety of reasons includ-
ing unrecognized disease, undertreated or
unresponsive to conventional therapy
asthma, recent exposure to triggers like
viruses, usually rhinovirus or allergens. AE
is dened as a worsening of asthma suf-
cient to require medical intervention and
usually administration of oral steroids
reected objectively as a fall of PEF, FEV1
?20% or 30%. This study was performed
to investigate the epidemics of exacerba-
tions in mites monosensitized asthmatics.
Seasonal variability of AE in such patients
could be used as a tool for a convenient
and preventive management. Non atopic
asthmatic patients were used as a control
group and this way we could understand
how important trigger is the allergen com-
paring to virus.
Method: We have studied, in our depart-
ment, during 20 years (from 1989 to 2009),
AE in patients suffering from these two
different phenotypes of asthma. Seasonal
variation of AE in each phenotype sepa-
rately and consequently comparison of
them were evaluated by applying the x2 -
test and the null hypothesis. The null
hypothesis is that there is not difference in
the number of AE occurred in each month
of the year.
Result: By applying the x2-test for 11 d.f
we obtained x2 = 84.319 which corre-
sponded to a P-value <0.0001. Therefore
the null hypothesis is not accepted and
there is statistical signicant difference in
the number of AE occurred during the
year (higher in spring and autumn
months).
By applying the x2-test for 11 d.f we
obtained x2 = 229.940 which corre-
Allergy 66, Suppl. 94 (2011): 482642  2011 John Wiley & Sons A/S
Table 1. For abstract 1497. Mite monosensitized patients
Month 1 2 3 4
Number of AE 32 41 58 59
Table 2. For abstract 1497. Non atopic patients
Month 1 2 3 4
Number of AE 249 239 245 237
sponded to a P-value <0.0001. Therefore
the null hypothesis is not accepted and
there is statistical signicant difference in
the number of AE occurred during the
year (higher in spring and winter months).
Comparison of both time distributions
revealed signicant difference between
them, x2 = 62.042 and P-value<0.0.
Conclusion: Our results showed that in
Greece during 20 years (from 1989 to
2009) AE are seasonal variable in non ato-
pic and mites monosensitized asthmatics
but different between them. The results
indicate that atopics react to viruses in a
different way comparing to non atopics.
1498
The reliability and validity of the Turkish
version of a childhood asthma control
test
Uysal Soyer, O1; Keskin, O2; Uzuner, N3; Yazicioglu, M4;
Kilic, M5; Artac, H6; Ozmen, S7; Can, D8; Zeyrek, D9;
Cokugras, H10; Sapan, N11; Aydogan, M12; Kuyucu, S13;
Inal, A14; Gurkan, F15; Orhan, F16; Yilmaz, O17; Bingol
Boz, A18; Tahan, F19; Cevit, O20; Sekerel, B1
1
School of Medicine, Pediatric Allergy and Asthma
Unit, Hacettepe University, Ankara, Turkey; 2Pediatric
Allergy Department, Gaziantep University, Turkey;
3
Pediatric Allergy Department, Dokuz Eylul University,
Turkey; 4Pediatric Allergy Department, Trakya
University, Turkey; 5Pediatric Allergy Department,
Ondokuz Mayis University, Turkey; 6Pediatric Allergy
Department, Selcuk University, Turkey; 7Pediatric
Allergy Department, Sami Ulus Childrens Hospital,
Turkey; 8Pediatric Allergy Department, Behcet Uz
Childrens Hospital, Turkey; 9Pediatric Allergy
Department, Harran University, Turkey; 10Cerrahpasa
Faculty, Pediatric Allergy Department, Istanbul
University, Turkey; 11Pediatric Allergy Department,
Uludag University, Turkey; 12Pediatric Allergy
Department, Kocaeli University, Turkey; 13Pediatric
Allergy Department, Mersin University, Turkey;
14
Pediatric Allergy Department, Cukurova University,
Turkey; 15Pediatric Allergy Department, Dicle
University, Turkey; 16Pediatric Allergy Department,
Karadeniz Technical University, Turkey; 17Pediatric
Allergy Department, Celal Bayar University,Turkey;
18
Pediatric Allergy Department, Akdeniz University,
Turkey; 19Pediatric Allergy Department, Erciyes
University, Turkey; 20Pediatric Allergy Department,
Cumhuriyet University, Turkey
Background: The management of asthma
is an important as well as difcult issue of
physicians daily practice particularly in
busy clinical settings. Childhood Asthma
Control Test (C-ACT) was created to iden-
tify asthma control levels in children aged
411 years. Our aim was to evaluate the
5 6 7 8 9 10 11 12
74 55 37 10 73 67 54 30
5 6 7 8 9 10 11 12
194 173 124 69 249 309 283 214
reliability, validity and responsiveness of of
C-ACT in a Turkish sample of children
with asthma.
Method: In this multicentre study; 368
children were enrolled. C-ACT was com-
pleted every month by parents and patients
who were evaluated in 3 visits with
2 month intervals. At each visit, physicians
interpret the control level and decided for
the treatment step as established in GINA
guidelines.
Result: The internal consistency reliability
of the Turkish version of C-ACT (C-ACT1
to C-ACT5) was found to be 0.82, 0.83,
0.82, 0.82 and 0.80, respectively. (reliability
statistics, Cronbach a). Test-retest reliabil-
ity was 0.71. There was signicant correla-
tion between C-ACT and physicians
assessment of asthma control at visit 1
(r = 0.65, P < 0.001).
Conclusion: Turkish version of C-ACT is
an accurate and reliable tool to evaluate
asthma control in children aged 411 years.
Its widespread use may facilitate appropri-
ate assessment of asthma control and may
lead to decrease the number of uncon-
trolled patients.
1499
Skin prick testing for aeroallergen sensi-
tivity is not necessary in very young chil-
dren with recurrent respiratory
complaints even in the case of high sus-
picion
Sahiner, U; Civelek, E; Yavuz, S; Buyuktiryaki, B;
Tuncer, A; Sekerel, B
Faculty of Medicine, Pediatric Allergy and Asthma,
Hacettepe University, Ankara, Turkey
Background: Sensitization to environmen-
tal allergens is a major risk factor for the
development of asthma, allergic rhinitis,
and atopic dermatitis. Early identication
of specic environmental allergies in chil-
dren may assist in medical and environ-
mental interventions in disease
management. The purpose of this study
was to perform an analysis of aeroallergen
sensitization rates for children who have
been referred to our clinic for SPT, speci-
cally focusing on infants and younger chil-
dren (< 2 years).
565

Method: The skin prick test results of the
children under two years of age presenting
with a complaint of recurrent respiratory
symptoms and highly suspicious of aeroal-
lergen sensitivity with or without eczema
between June 2008 and June 2010 to the
largest referral center of the country were
evaluated. The aeroallergen skin prick test-
ing is not a routine procedure in the clinic
and only performed in the case of suspi-
cion to a specic allergen.
Results: A total of 3783 children under
2 years of age were examined in the clinic
during the study period and among them
701 children (male/female: 1.99) with a
median (inter quartile) age of 1.10 (0.71-
1.57) years were selected to be SPTed with
aeroallergens (Stallergenes; Antony,
France). The type of aeroallergen, number
of the SPTed children (n) and % of the
SPT sensitizations among the ones tested
are as follows: Dermatophagoides farinae
(638) 4.2%; Dermatophagoides pteronyssi-
nus (534) 3.0%; Cat (35) 2.9%; Festuca
pratensis (144) 1.4%; Poa mix (144) 1.4%;
Cynodon dactylon (142) 1.4%; Phleum pra-
tense (142) 1.4%; dog (141) 0.7%; Betula
alba (140) 0.7%.
Conclusion: In children less than 2 years of
age sensitization rates to common aeroal-
lergens are very low (< 5%) and it
appears that testing young children for ae-
566
Poster Session 63 Pediatric asthma: clinical assessment and therapy
roallergens sensitivity is not necessary even
in the case of high suspicion.
1500
Serum vitamin D levels in wheezy infants
Durmaz, C1; Asilsoy, S1; Usta Guc, B1; Anarat, R2
1
Baskent University Hospital, Pediatric Allergy, Turkey;
2
Baskent University Hospital, Biochemistry, Turkey
Background: Epidemiologic studies have
shown that low serum 25(OH)D levels are
associated with increased risk of lower
respiratory tract infections in young chil-
dren. Almost all viral respiratory tract
infections occur between November and
March, months that also are associated
with vitamin D deciency. Vitamin D
recently has been shown to have an impor-
tant role in the immunoregulation espe-
cially in innate immunity and production
of antimicrobial peptides. In this study, we
evaluated the relationship between serum
25(OH)D levels and recurrent respiratory
tract infections with wheezing in young
children.
Method: We analysed 25(OH)D serum lev-
els in 60 wheezy infants, aged 636 months
with three or more viral wheezing episodes
and 40 healthy controls at the same age.
The study took place in the Department of
Allergy 66, Suppl. 94 (2011): 482642  2011 John Wiley & Sons A/S
Pediatric Allergy of Baskent University
between November and March, months
that viral wheezings were common. We
categorized vitamin D levels into decient
(20 ng/ml), insufcient (2129 ng/ml) and
sufcient (30 ng/ml) based on previous
recommendations.
Result: The mean serum 25(OH)D concen-
tration in wheezy infants (30.1 7.6 ng/
ml) was lower than healthy controls
(33.9 9.8 ng/ml), (P < 0.05). Vitamin D
deciency was 7/60 (%12) in wheezy
infants and 2/40 (%5) in healthy controls,
vitamin D insufciency was 21/60 (%35) in
wheezy infants and 13/40 (%32) in healthy
controls (P > 0.05). Serum vitamin D lev-
els were signicantly associated with age,
period of breastfeeding and maternal cloth-
ing in wheezy infants (P < 0.05).
Conclusion: Vitamin D insufciency is pre-
valent among young children in our region.
According to the previous studies support-
ing the role of vitamin D in innate immu-
nity and antiviral defence, we suggest that
it might be particularly important in
wheezy children. New guidelines for ade-
quate sun exposure and supplemental vita-
min D in children are needed.

Allergy and asthma: worldwide experiences, risk factors and
1501
Pediatric asthma control in Japan
Sato, K; Suzuki, S; Nezu, Y; Matsura, T; Watanabe, H;
Nishimuta, T
Department of Pediatrics, National Shimoshizu
Hospital, Chiba, Japan
Background: Control based asthma man-
agement was recommended also in chil-
dren, but not investigated enough,
especially in preschool child, because the
diagnosis and the evaluation are not easy.
Some validated asthma control question-
naire for children were reported. But ques-
tionnaires of the foreign country are not
always useful because of differences in
socio-economy and medical insurance sys-
tem. We investigated Asthma control in
children using original questionnaire, the
JPAC (Japanese Pediatric Asthma Control
program).
Method: 1009 subjects with doctor diag-
nosed asthma, 115 years old, regular out-
patients of thirty three hospitals and clinics
in Chiba prefecture in Japan were
recruited. The JPAC questionnaire used
for 4 to 15 years old children has 5 items,
wheezing frequency in 4 weeks. Frequency
of breathing difculties in 4 weeks, Fre-
quency of sleep disturbance in 4 weeks, fre-
quency of exercise induced respiratory
symptoms, and frequency of rescue use of
beta agonist. The questionnaire used for 1
to 3 years old children was arranged JPAC
for younger children, added one items
about frequency of cough without fever for
4 weeks. Each question was 3 points full
mark, 15 points and 18 points means best
control, respectively. Asthma control was
evaluated according to JPAC score in three
degree, total controlled, well controlled,
not well controlled. The changes in asthma
control and treatment were also examined
in some subjects agreed for investigation of
two consecutive visits.
Result: In 4 to 15 years old children,
34.1% were totally controlled, 38.0% were
well controlled, and 27.9% were not well
controlled. In 1 to 3 years old children,
27.6% were totally controlled, 44.8% were
well controlled, and 27.6% were not well
controlled. Treatments of poorly controlled
Allergy 66, Suppl. 94 (2011): 482642  2011 John Wiley & Sons A/S
Poster Session 64
epidemiology
children were changed only in 15.4% of
younger and in 21.6% of elder children.
Conclusion: Pediatric asthma patients in
Japan are well controlled. But pediatric
asthma management based on the control
is not well established, yet.
1502
Dietary fat intake and asthma in Korean
children
Seo, J1; Kim, H1; Kwon, J1; Hong, S1; Yu, J2; Oh, S3;
Lee, S4; Kim, H5; Kim, B6; Shin, Y7
1
Pediatric Allergy Department, Asthma Atopy Center,
Asan Medical Center, University of Ulsan College of
Medicine, Seoul, Republic of Korea; 2Department of
Pediatrics, Asthma Atopy Center, Asan Medical Center,
University of Ulsan College of Medicine, Seoul,
Republic of Korea; 3Department of Food and Nutrition,
College of Human Ecology, Kyung Hee University,
Seoul, Republic of Korea; 4Department of Pediatrics,
Hallym University Sacred Heart Hospital, University of
Hallym College of Medicine, Seoul, Republic of Korea;
5
Department of Pediatrics, Pediatric Asthma & Allergy
Center, Inje University Sanggye Paik Hospital, Seoul,
Republic of Korea; 6Department of Pediatrics, Pediatric
Asthma & Allergy Center, Inje University Haeundae
Paik Hospital, Pusan, Republic of Korea; 7Department of
Psychiatry, Severance Hospital, College of Medicine,
Yonsei University, Seoul, Republic of Korea
Background: The prevalence of allergic dis-
eases has risen over the past few decades,
mostly due to environmental and life style
changes including diet. Among dietary fac-
tors, fatty acid intake has been associated
with asthma.
Method: Objectives: To investigate the
association of fat intake with asthma in
Korean school children aged 89 years old.
Methods: Subjects were 1239 children from
24 elementary schools in Korea. The risk
of allergic disease was measured using the
Korean version of the International Study
of Asthma and Allergies in Childhood and
dietary intake was measured by semi-quan-
titative food frequency questionnaire.
Intake of total fat, cholesterol, saturated
fatty acid, unsaturated fatty acid including
n-3 and n-6 polyunsaturated fatty acid.
Result: The rate of wheeze ever in life was
24.7% and the rate of current wheeze
11.4%. The diagnosis and treatment rate
of asthma was reported in 10.4%, 4.2%
respectively. Total fat intake was positively
associated with an increased risk of wheeze
ever (aOR = 1.21 95% CI = 1.0541.390)
and current wheeze (aOR = 1.195, 95%
CI = 1.0011.427). Monounsaturated and
polyunsaturated fatty acid intakes were
positively associated with current wheeze
(aOR = 1.190, 95% CI = 1.0081.405,
aOR = 1.215, 95% CI = 1.0271.436
respectively) and asthma treatment
(aOR = 1.288, 95% CI = 1.0161.634,
aOR = 1.247, 95% CI = 1.0051.548).
There was no association between choles-
terol intake and wheeze or asthma. There
was no association with atopy or bronchial
hyperresponsiveness.
Conclusion: Increased consumption of total
fat and fatty acid may affect the develop-
ment of wheeze and asthma, but not atopy
and bronchial hyperresponsiveness, in Kor-
ean children. This study was supported by
a grant of Childrens Health and Environ-
ment Research of Ministry of Environ-
ment, Republic of Korea.
1503
Association of obesity and asthma in
Korea
Park, Y1; Lim, S2; Yoo, K3
1
College of Medicine, Internal medicine, Hallym
University Seoul, Republic of Korea; 2Kangbuk
Samsung Hospital, Sungkyunkwan University School of
Medicine, Internal medicine, Seoul, Republic of Korea;
3
KonKuk University, Internal medicine, Seoul, Republic
of Korea
Background: Although several studies have
suggested a relationship between asthma
and obesity, debates still remain. This
study was to determine whether obesity is
associated with asthma and atopy in Kor-
ean adults.
Method: This study was case control multi-
center study in Korea between January
2007 and June 2008. The subject included
who had experienced chronic cough symp-
tom. We examined body mass index (BMI)
for measurement of obesity, pulmonary
function test, metacholine provocation test
for airway hyperresponsiveness (AHR),
skin prick test for atopy, blood eosinophil
and serum IgE.
Result: A total 1022 subjects were
included. FEV1/FVC was signicantly
reduced in the group with obesity
(P-value = 0.03), but FEV1, FVC were
567

not different from the group with normal
BMI. AHR was not related with obesity
(P-value = 0.06), and atopy prevalence
was signicant higher in non obese patients
(P-value = 0.0). There was no difference
in Ig E and Eosinophil count between
obese and non obese group.
Conclusion: In this study, we could not
found relationship between obesity and
asthma. The non-obese group appears to
have more atopy. The relationship between
obesity and asthma and atopy need further
investigation.
1504
The effect of obesity and changes in
weight on asthma symptoms in school
children
Kwon, J1; Kim, H1; Kim, B2; Yu, J1; Lee, S3; Hong, S1;
Kim, H4
1
Asan Medical Center, University of Ulsan College of
Medicine, Pediatrics, Seoul, Republic of Korea; 2Inje
University Haeundae Paik Hospital, Pediatrics, Busan,
Republic of Korea; 3Hallym University Sacred Heart
Hospital, Pediatrics, Seoul, Republic of Korea; 4Inje
University Sanggye Paik Hospital, Pediatrics, Seoul,
Republic of Korea
Background: Prevalence of asthma has
increased concurrently with a rise in over-
weight in children, but it is unclear
whether body mass index (BMI) has an
effect on the risk of childhood asthma
development.
Method: We performed a longitudinal
study with 1 338 children aged 7 years and
followed up for 4 years. We measured each
childs body physics and investigated sensi-
tization to allergens, pulmonary function,
and bronchial hyperresponsiveness (BHR)
every 2 years. Questionnaires with asthma
symptoms, environmental and social fac-
tors were reported by the parents at each
follow-up.
Result: Asthma symptoms such as ever
wheeze and wheeze in previous 12 months
were more prevalent in overweight children
than normal (32.0% versus 23.2%,
P = 0.001 and 15.3% versus 11.0%,
P = 0.004). FEV1/FVC was signicantly
decreased in overweight children compared
to normal (P < 0.001) and we found nega-
tive correlation between BMI and FEV1/
FVC (r2 = )0.103, P < 0.001). Presence
of BHR and atopic sensitization were not
signicantly different between overweight
and normal children. Among the children,
5.4% of them became overweight during
4 years, 6.7% decreased their BMI, and
18.8% showed persistently high BMI. Chil-
dren who were persistently overweight or
who became overweight during 4 years of
follow-up did not have signicant risk of
development of asthma symptoms, BHR,
and atopy. CRP was signicantly high in
children who had persistent overweight
568
Poster Session 64 Allergy and asthma: worldwide experiences, risk factors and epidemiology
during 4 years compared to who had per-
sistently normal BMI.
Conclusion: Overweight was associated
with wheeze symptoms and pulmonary
function, but not with BHR or atopy.
Changes of weight status during school-age
did not have an effect on asthma symp-
toms, BHR and atopic sensitization. Over-
weight has an association with
inammation, which may be presented by
CRP.
1505
Epidemiology of allergic diseases among
the adult population of the CIS-countries
Sepiashvili, R
Institute of Immunophysiology, Allergology and Clinical
Immunology, Moscow, Russian Federation
Background: In this study there has been
analyzed the data from epidemiological
studies on the prevalence of allergic dis-
eases (AD) among the adult population in
the territory of the CIS countries (CIS-C)
over the past 510 years.
Materials and methods: For the analysis
there has been used the literature data; the
results of studies of ISAAC and ARIA
programs, obtained from the National
Societies of Allergology and Immunology;
statistical reports of the Republic Minis-
tries of Health and Medical facilities.
Results: It has been found that the struc-
ture of AD of CIS-C is dominated by:
bronchial asthma (BA), allergic rhinitis
(AR), atopic dermatitis (AD), pollinosis
(P) and allergic conjunctivitis (AC). AR,
on average, has been diagnosed at the 17
21% (from 5.8 to 33.3%) adults. AR, on
attendance data, suffers only 0.10.4% of
the population. It has been stated that 7%
of 48.3% of the adult population suffer
from BA. The highest incidence of BA
among adults is reported in Armenia, Bela-
rus, Moldova, Ukraine, Kyrgyzstan, Rus-
sia and Tajikistan. The actual incidence of
the adult population reaches 2348%. In
this case, the diagnosis of asthma institu-
tions of practical public health has been
recorded in adults less than 1%. The aver-
age incidence of AD in the CIS-C in adults
was 6 28.7%. The high prevalence of AD
is typical for Ukraine, Tajikistan, Kazakh-
stan, Kyrgyzstan and Belarus. In Tajiki-
stan, Belarus, Kazakhstan and Armenia, a
signicant number of patients with P has
been revealed, the incidence among adults
reaches 15.729.3%. Among all CIS-C AK
most often occurs in Armenia, Tajikistan,
Kazakhstan, Azerbaijan and Georgia. For
example, in Armenia, there has been estab-
lished a signicant incidence of AK among
adults, the disease is diagnosed in 13.6
32.9% cases. In Kazakhstan and Tajiki-
Allergy 66, Suppl. 94 (2011): 482642  2011 John Wiley & Sons A/S
stan, there has been also revealed wide-
spread AK among adults (17.523.1%).
For other CIS-C this disease is not socially
signicant. Analysis of epidemiological
data has shown that prevailed patients
with mild disease, whereas in the structure
of AD, registered on negotiability, domi-
nated the moderate disease course.
Conclusion: Epidemiological studies have a
great scientic and practical importance
that provide reliable information on the
prevalence, the structure and the severity
of AD. Monitoring of the true prevalence
of AD indicates hypodiagnostics of the all
forms of AD in the CIS-C, related to the
underestimation of the mild forms of the
disease.
1506
Farm-related exposures are associated
with a persistent lower prevalence of
sensitisation to pollen
Elholm, G1; Omland, 2; Schlunssen, V3; Doekes, G4;
Basinas, I3; Hjort, C5; Milvang Grnager, P6; Sigsgaard, T3
1
Department of Environmental and Occupational
Medicine, University of Aarhus, Aarhus, Denmark;
2
Alborg Hospital, Arhus University Hospital, Clinic of
Occupational Medicine, Alborg, Denmark; 3Department
of Environmental and Occupational Medicine, Aarhus
University, Aarhus, Denmark; 4Institute for Risk
Assessment Sciences, Division of Environmental
Epidemiology, Utrecht University, Utrecht, Netherlands;
5
Department of Quality and Research, Regional
Hospital of Viborg, Skive, Kjellerup, Viborg, Viborg,
Denmark; 6Research ALK Abello, Denmark
Background: Farmers are exposed to a
wide range of allergens. However the prev-
alence of atopic sensitization and atopic
asthma has been reported to be low in
farmers. We aim to study the relation
between farm-related dust and endotoxin
exposures and the persistence, loss, or new
incidence of atopic sensitisation over time
in The Danish Farming Cohort (SUS).
Method: The SUS cohort consists of 1964
young farmers and 407 non-farming male
controls. The participants were examined
rst during the SUS study (19921994) and
52% of the cohort was re-investigated dur-
ing the follow-up SUS12 study (2006
2008). The participants had a blood sample
taken at both baseline and follow-up for
analysis of specic IgE (ADVIA Centaur,
ALK Abello ) to allergens from cat (Felis
domesticus), birch (Betula verrucosa), grass
(Phleum pratense) and HDM (Dermato-
phagoides pteronyssinus). Personal average
weekly exposure to dust and endotoxin
during the follow-up period was estimated
by exposure modelling with an internal job
exposure matrix based on farm type and
more than 500 personal inhalable dust
measurements.
Result: Sensitisation was dened as
sIgE0.35 kU/l for at least one of the four
allergens. Sensitisation status was grouped
Poster Session 64 Allergy and asthma: worldwide experiences, risk factors and epidemiology
Table 1. For abstract 1506. Changes in IgE sensitisation to common allergens
Sensitisation
Never
()) ())
n = 841
Current farmer n(%)* 348 (79.0%)
Ever farmer n(%)* 375 (75.2%)
Never farmer n(%)* 106 (66.3%)
Dust exp (mg/m3/year) # 6.8 (068.4)
Endotoxin exp 1989 (029422)
(EU/m3/year) #
Grass, birch, cat, or HDM.
: Median (min max).
*: (n) may vary due to missing values.
#: work years calculated assuming a work week of 40h/w/y.
into never sensitised, only sensitised at
baseline, only sensitised at follow-up and
sensitised at both baseline and follow-up.
As shown in Table 1, current farmers were
more likely not to be sensitised at both
time-points (79%) compared to never
farmers (66%). Never farmers (25%) were
more likely to be sensitised at both base-
line and follow-up than current farmers
(12%). The median levels of exposure to
both dust and endotoxin were associated
to sensitisation status, with the highest
exposure level among subjects loosing sen-
sitisation in the follow up period and the
lowest exposure level among subjects sensi-
tised at both baseline and follow-up. When
analysing sensitisation rates to each of the
four allergens separately, the exposure
dependent differences in sensitisation were
mainly driven by differences in pollen
(birch and grass) sensitisation.
Conclusion: These analyses support the
hypothesis that farming exposure has a
persistent protective effect on sensitisation.
Furthermore the results in this study sug-
gest that this protective effect is mainly
seen for pollen sensitisation.
1507
Epidemiological study of the causes of
allergic and respiratory diseases in Rus-
sia
Pavlova, K
Clinical and Epidemiological Trials Department, SSC
Institute of Immunology, Moscow, Moscow, Russian
Federation
Background: Understanding of the patho-
genesis and management of bronchial
asthma (BA) and nasal polyps (NP) remain
an actual problem. This study is a part of
an GA2LEN epidemiological program, in
order to detect of possible causes that
impact to BA and NP development.
Methods: The study consisted of two parts.
In the rst part of the study were selected
5000 people from the resident of Moscow
Allergy 66, Suppl. 94 (2011): 482642  2011 John Wiley & Sons A/S
Sensitisation Sensitisation Sensitisation P-value
Follow-up Baseline Always
()) (+) (+) ()) (+) (+)
n = 62 n = 30 n = 181
22 (5.1%) 14 (3.2%) 52 (11.9%) 0.008
25 (5.0%) 13 (2.6%) 85 (17.1%)
11 (6.9%) 3 (1.9%) 40 (25.0%)
5.5 (070.9) 8.3 (038.7) 3.91 (041.7) 0.0009
1192 (031677) 2032 (016487) 983 (017159) 0.0004
aged 18 to 75 years by a cross section
through by a representative simple random
sampling. These subjects were invited to
study and responded to the questionnaire
to detect the allergic and respiratory dis-
eases. In the second part of the study 300
individuals responded to the Screening
questionnaire and indicated they are happy
to take part in further research were
selected. They were divided to three
groups: 90 people with BA, 90 people with
chronic rhinosinusitis (CRS), 90 people
with a combination of BA and CRS and
one control group (30 people). All individ-
uals completed the questionnaire GA2LEN
Follow up, were examined allergist, otolar-
yngologist, performed general clinical
examination, skin prick tests for a standard
set of allergens, measured total IgE and
specic IgE to Alternaria, Aspergillus, Cla-
dosporium and superantigens S. aureus A,
B and TSS and spirometry before and after
salbutamol 200g.
Results and discussion: We received 2 500
questionnaires, 1 523 were completed and
977 were empty. In 54.3% (of 1523
responders) we assumed the presence of
any allergic disease. 7.1% patients (of 1
523 responders) have BA, and 15.3% peo-
ple have possibility of BA. As a result of
complex examination 7 patients (of 300)
was rst diagnosed BA, 35 persons
(19.4%) with previously diagnosed BA
hadnt the adequate asthma therapy, which
indicates that low status and no control of
asthma. Increased specic IgE to superanti-
gens S. aureus was typical for patients with
CRS (as with the presence of NP, with or
without NP), not previously treated with
appropriate antimicrobial therapy. The
presence of specic IgE to superantigens
S. aureus did not correlate with any
increase of total IgE, nor to the presence
of sensitization to atopic allergens nor with
the severity of the main disease.
1508
Life style and environmental as they
relate to allergic disease in rural Poland
Bialoszewski, A1; Kapalczynski, W2; Samoliski, B1;
Lusawa, A1; Walkiewicz, A1; Samel-Kowalik, P1
1
Department of Prevention of Environmental Hazards
and Allergology, Medical University of Warsaw,
Warsaw, Poland; 2University of Louisville School of
Medicine, Louisville, United States
Background: The ECAP 2006 study was
carried out with the help of 22 700 partici-
pants in nine regions of Poland. Nearly a
quarter of these subjects underwent subse-
quent medical evaluation. The results of
the study show that the prevalence of aller-
gic disease in Poland is among the highest
in the world. In this study, we wanted to
detail the results of the research
ECAP2006.
Method: ECAP 2010 constituted the con-
tinuation of previous studies that employed
ECRHS and ISAAC standards. The study
was carried out in rural Poland and
included 355 participants. The question-
naire contained 53 questions pertaining to
the habitat and lifestyle of the participant.
The clinical portion included a qualifying
exam, spirometry and skin prick testing.
Result: Our study demonstrated that the
incidence of allergy in adults is 2.019 times
greater (95% CI: 1.2853.173, P = 0.002)
than in children. There was no difference
observed between genders. Better educated
subjects suffered more often from skin
allergies (P = 0.044). Participants inhabit-
ing smaller dwellings (below 80 m2) had an
approximately 20% higher chance of hav-
ing any type of allergy (P = 0.007) and a
15% higher chance of having a skin allergy
(P = 0.003). Subjects who owned a well
had an 8% lower chance of skin allergy
(P = 0.024) than those with access to the
municipal water supply. Owning a septic
tank was associated with a 13%
(P = 0.014) reduction in the incidence of
skin allergy. Those who were breast fed as
infants had a 17% (P = 0.009) lower
chance of developing skin allergies. The
largest difference was observed between the
groups who were treated with antibiotics
during their childhood. Those who had
taken antibiotic therapy six or more times
had a 17% increased chance of suffering
from any type of allergy than those who
had taken antibiotic therapy up to six
times.
Conclusion: The data shows a strong rela-
tionship between environmental and life-
style factors and the incidence of allergy.
The strongest risk factors for allergy were
frequent antibiotic treatment during child-
hood and not being breastfed as an infant.
Surprisingly, owning a well or a septic tank
was correlated with a decreased risk of
skin allergy. Gender did not seem to have
569

an inuence on the incidence of allergy.
Age, however, was demonstrated to have a
strong inuence on the incidence of allergy,
and underlines the importance of the need
to study factors that inuence our health
during childhood.
1509
Prevalence of asthma in Western
Hungary from 2006 to 2009
Soti, L1; Petz, Z1; DuBuske, L2
1
Allergy-Asthma Outpatient Clinic, Szombathely,
Hungary; 2Immunology Research Institute of New
England, Gardner, MA, United States
Background: This investigation assesses the
prevalence of asthma in various age ranges
including the distribution of patients based
on their gender in Western Hungary. The
data was collected at the Allergy Outpa-
tient Clinic of Western Hungary, a diag-
nostic centre which receives patients from
a region with a population of 263 251.
Method: Patients with newly diagnosed
asthma were included, from infants to
96 years old from among 30 335 patients
seen from 2006 through 2009. Asthma was
assessed by a specialist by skin prick tests
and/or allergen specic IgE tests and spi-
rometry.
Result: The average incidence of asthma
was 112% of the total population with the
maximum incidence in the age group of 5
to 6 years old, with boys having twice the
incidence of girls in this group. The overall
peak incidence of asthma occurred in chil-
dren from 4 to 7 years old. Male children
show a subsequent decline in their asthma
incidence rates, the male incidence crossing
that of the female children at age 15.
Females have a greater incidence than
males from age 15 until age 60 and above,
with the overall incidence of asthma
declining in the older age groups for both
sexes.
Conclusion: Childhood asthma remains a
problem in Western Hungary, especially
among young boys. The shift to female
dominance in asthma prevalence at age 15
suggests hormonal factors contribute to the
development of asthma, with female pre-
dominance remaining through the older
age groups.
1510
Analysis of the quality of life among
asthmatic children in Serbia
Krivokapic, T; Zivkovic, Z; Radic, S; Kalaba, Z;
Milanovic, V
Center for Childrens pulmonology, pulmonology and
allergology, Belgrade, Serbia
Background: The aim of our study was to
detect is there any inuence of symptoms
570
Poster Session 64 Allergy and asthma: worldwide experiences, risk factors and epidemiology
and objective parameters (lung function)
on quality of life children with asthma.
Method: Group of 40 children (23 female,
17 male), age 718 with persistent moder-
ate asthma were hospitalized after asthma
exacerbation. We measured spirometry and
PAQLQ (the Junipers Pediatric Asthma
Quality of Life Questionnaire). The evalua-
tion of the quality of life was based on a
questionnaire with 23 items divided in
symptoms, activities and emotions. After
the urgent treatment we reevaluated their
lung function test, PAQLQ at 1st 7th and
30th day.
Result: Symptoms were progressively
reduced from 1st to 7th and 30th day from
the exacerbation, also lung function was
improved, especially in small peripherial
airways. The statistically difference FEV1
and Tiffeneau index 1st day and 7th day
was statistically signicant, also 1st day
and 30th day P < 0.001. There was no
statistically signicant difference between
7th and 30th day in FEV1 and Tiffeneau
index. PAQLQ Scor 1st day was 4.21
1.09, 7th day PAQLQ scor was 4.74
1.29, 30th day PAQLQ score was
5.19 0.96. The difference in PAQLQ
score between 1st and 30th day
(P = 0.001), and score between 7th and
30th day wasnt statistically signicant
(P = 0.382).
Conclusion: We found the difference in
PAQLQ and spirometry parameters 1st
day and 7th day, also 1st and 30th day.
Quality of life questionnaire is an impor-
tant parameter for measuring subjective
impact of asthma on someones condition
and daily life. This analysis helps us to
improve and complete physical examina-
tion and eventually correct prophylaxis. In
children with persistent asthma manage-
ment of asthma must include objective as
well as subjective measurement of control.
1511
Epidemiological survey of adult asthma
in the Republic of Macedonia
Stoleski, S1; Karadzinska-Bislimovska, J1; Minov, J1;
Mijakoski, D2; Risteska-Kuc, S2; Marsenic, M2;
Milkovska, S2
1
Department for Cardiorespiratory Functional
Diagnostics, Institute for Occupational Health of R.
Macedonia, WHO Collaborating Center, GA2LEN
Collaborating Center, Skopje, Macedonia, FYROM;
2
Institute for Occupational Health of R. Macedonia,
WHO Collaborating Center, GA2LEN Collaborating
Center, Allergy Center, Skopje, Macedonia, FYROM
Aim of the study: To evaluate the asthma
prevalence in a general population sample
and in subjects with respiratory symptoms
in R. Macedonia.
Methods: We performed a cross sectional
study including 722 subjects from six cen-
ters in R. Macedonia (283 males and 439
Allergy 66, Suppl. 94 (2011): 482642  2011 John Wiley & Sons A/S
females, mean age 39.6 13.5). Evalua-
tion of the examined subjects included
completion of an interviewer-led question-
naire designed according to the ECRHS
screening questionnaire, as well as ques-
tions about family history of asthma and
allergic diseases, current smoking and
smoking history, environmental and work-
place exposure, medication use, and skin
prick tests to 13 common inhalant aller-
gens. Spirometry and histamine challenge
were performed in the examined subjects
aged 2044 who reported symptoms sug-
gestive of asthma.
Result: Prevalence of asthma in the age
group 2044 was 5.4%, 6.4% in males and
4.9% in females, which translated to
approximately 100.000 subjects with
asthma in R. Macedonia. The highest
asthma prevalence was registered in Skopje
(6.3%), and the lowest one in Dojran
(3.3%). Allergic asthma was detected in
60%, and non-allergic asthma in 40% of
the subjects with asthma. Sensitization to
Dermatophagoides pteronyssinus and goose-
foot pollen were the most important com-
mon allergens in the subjects with allergic
asthma. We found asthma/rhinitis associa-
tion in 70% of the subjects with asthma
that conrmed the united airway disease
concept. In the majority of subjects with
asthma accompanied by rhinitis, asthma
development was preceded by rhinitis. We
found signicant association of asthma
with atopy and family history of asthma.
Allergic rhinitis was detected in about half
of the atopic subjects, while asthma was
detected in about 15% of the atopic sub-
jects. Paternal asthma was reported by half
of the subjects with asthma. Association
between asthma and other endogenous fac-
tors, such as sex, ethnic group, and family
history of allergic diseases, was non-signi-
cant. We found signicant joint effect of
some environmental factors (urban resi-
dence, active smoking, exposure to trafc
pollution, and green plants outdoors) on
asthma development. Prevalence of work-
related asthma was 1.4%, (24.7% of the
subjects with asthma), being the highest
among pharmaceutical, textile and chemi-
cal industry workers.
Conclusion: Our results suggest that the
asthma prevalence must be taken in con-
sideration in allergologic evaluation of the
subjects with allergic diseases and respira-
tory symptoms.
Poster Session 64 Allergy and asthma: worldwide experiences, risk factors and epidemiology
1512
The Portuguese national asthma survey
what is the prevalence of asthma?
Sa e Sousa, A1; Morais de Almeida, M2; Filipe Azevedo,
L1; Carvalho, R3; Todo-Bom, A4; Loureiro, C5; Bugalho
de Almeida, A6; Almeida Fonseca, J7
1
Faculdade de Medicina da Universidade do Porto,
Biostatistics and Medical Informatics Department,
Porto, Portugal; 2Imunoalergologia, Hospital CUF-
Descobertas; Sociedade Portuguesa de Alergologia e
Imunologia Clnica, Portugal; 3GBN, Estudos de
Mercado, Portugal; 4Sociedade Portuguesa de
Alergologia e Imunologia Clnica; Imunoalergologia,
Hospitais da Universidade de Coimbra, Portugal;
5
Imunoalergologia, Hospitais da Universidade de
Coimbra; Sociedade Portuguesa de Alergologia e
Imunologia Clnica, Portugal; 6Clnica Universitaria de
Pneumologia, Faculdade de Medicina de Lisboa;
Comissao Acompanhamento Programa Nacional
Controlo Asma, Portugal; 7SBIM/CINTESIS, Faculdade
Medicina Universidade Porto;Imunoalergologia,
Hospital CUF/Hospital S. Joao, Porto, Portugal
Background: We aimed to estimate the
prevalence of asthma symptoms in Portu-
gal.
Method: Nationwide, cross-sectional list-
assisted random-digit-dialing telephone
interview survey (rst-phase of the Portu-
guese National Asthma Survey). The ques-
tionnaire was based on the GA2LEN
survey. Current asthma was dened as self-
reported asthma and one of 3 symptoms in
the last 12 months: wheezing, awaking with
breathlessness or having an asthma attack.
Data analysis was weighted having into
account age and sex distribution of the lat-
est national census (2001). Interviews were
conducted between March and May of
2010.
Result: Of the 6 003 participants, 57.3% (3
438) were females; 11.9% (716) were chil-
dren, 51.7% (3 104) adults (between 18
and 65 years old) and 36.3% (2 178)
elderly; 65.1% (3 907) had up to 9 years of
school education, 19.6% (1175) between 9
and 12 years and 12.3% had more than
12 years; 6.7% (405) belonged to the high,
68.9% (4 137) to medium and 21.3% (1
280) to low socioeconomic status; 36.3%
(2 179) had normal weight, 30.4% (1 825)
were overweight and 11.8% (710) obese.
The prevalence of current asthma in the
Portuguese population was 6.8% (95% CI
6.07.7) and the life-time prevalence was
10.5% (95% CI 9.511.6). From these,
asthma onset occurred 72.8% in childhood,
25.5% in adult age, and 1.7% in elderly
age. The prevalence of asthma symptoms
in the last 12 months was: 18.1% for
wheezing, 35.6% for nocturnal symptoms
and 3.2% for asthma attack. The preva-
lence of current asthma was 7.2% in
females and 6.3% in males; according to
age groups it was 7.2% (95% CI 5.49.5)
in children, 6.3% (95% CI 5.37.5) in
adults and 8.0% (95% CI 6.79.5) in the
elderly. Current asthma distribution by
BMI was 6.1% normal weight, 6.6%
Allergy 66, Suppl. 94 (2011): 482642  2011 John Wiley & Sons A/S
underweight, 6.4% overweight, 8.7% in
obese. Concerning school education
groups, it was 7.1% in the rst 9 years,
7.4% in 9 to 12 years and 4.5% in the
more than 12 years. Regarding the asth-
matic population, 52.3% (95% CI 45.8
58.6) had symptoms of allergic rhinitis.
Conclusion: For the rst time a nationwide
survey estimated the prevalence of asthma
in Portugal 695,000 (6.8%) have current
asthma and more than one million (10.5%)
had asthma during their lives.
1513
The Portuguese national asthma survey
Over 40% of asthma patients are con-
trolled
Sa e Sousa, A1; Morais de Almeida, M2; Filipe Azevedo,
L3; Carvalho, R4; Todo-Bom, A5; Loureiro, C6; Robalo-
Cordeiro, C7; Bugalho de Almeida, A8; Almeida
Fonseca, J1
1
Faculdade de Medicina da Universidade do Porto,
Biostatistics and Medical Informatics Department,
Porto, Portugal; 2Imunoalergologia, Hospital CUF-
Descobertas; Sociedade Portuguesa de Alergologia e
Imunologia Clnica, Portugal; 3Faculdade de Medicina
da Universidade do Porto, Biostatistics and Medical
Informatics Department; CINTESIS, Porto, Portugal;
4
GBN, Estudos de Mercado, Portugal; 5Sociedade
Portuguesa de Alergologia e Imunologia Clnica;
Imunoalergologia, Hospitais da Universidade de
Coimbra, Portugal; 6Imunoalergologia, Hospitais da
Universidade de Coimbra; Sociedade Portuguesa de
Alergologia e Imunologia Clnica, Portugal;
7
Pneumologia, Hospitais da Universidade de Coimbra;
Sociedade Portuguesa de Pneumologia, Portugal;
8
Clnica Universitaria de Pneumologia, Faculdade de
Medicina de Lisboa; Comissao Acompanhamento
Programa Nacional Controlo Asma, Portugal
Background: We aimed to estimate the
proportion of asthmatic patients with con-
trolled disease in Portugal.
Method: Second phase of a nationwide,
cross-sectional list-assisted random-digit-
dialing telephone interview survey. Partici-
pants identied as having asthma in
rst-phase of the survey that addressed
prevalence of asthma were re-interviewed.
The questionnaire included ATAQ,
CARAT, MiniAQLQ, MiniRQLQ. Con-
trolled asthma was dened based on CAR-
ATs global and factor2 scores: global
score superior to 24 or factor2 score 16 or
higher. Interviews were conducted between
June and October of 2010.
Result: The total participants was 559,
60% (336) were females; 20% (110) chil-
dren, 50% (282) adults and 30% (167)
elderly; 64% (357) had up to 9 years of
school education, 22% (121) between 9
and 12 years and 12% (66) had more than
12 years; 8% (43) belonged to the high,
78% (422) to medium and 17% (92) to low
socioeconomic status and 40% (226) had
normal weight, 8% (44) underweight, 27%
(149) overweight and 18% (103) obese.
Regarding asthma symptoms, 39% (155)
had daytime symptoms, 30% (119) had
limitation of daily life activities, 32% (128)
had nocturnal symptoms, 4% (18) needed
reliever treatment. In relation with their
asthma, in the last 12 months, 68% (278)
had at least one medical visit, 25% (102)
assessed lung function and 23% (93) went
to an ER. Asthma was controlled in 57%
(95% CI 5262) of the participants and
44% (95% CI 3948) achieved clinical
GINA goals. Of the individuals with
uncontrolled asthma 88% (95% CI 8393)
had a decient awareness of their asthma
control level. Controlled asthma was more
frequent in males (71% versus 46% in
females, P < 0.001), in high socioeco-
nomic status (82% versus 60%in medium
and 31 in low, P < 0.001) and in partici-
pants with more years in school (49% in
the rst 9 years, 69% in 9 to 12 years and
73% in the more than 12 years,
P < 0.001). Asthma control was also
lower (P < 0.001) in the elderly (36%)
compared to children (68%) and adults
(63%). Concerning BMI, proportion of
controlled asthma was 68% in under-
weight, 61% in normal weight, 52%in
overweight and 51%in obese.
Conclusion: In Portugal, the proportion of
asthmatic patients with controlled disease
is very high compared to previous studies
in other countries. One may speculate that
the national program on asthma control
probably had some inuence on these posi-
tive results. However, as this is the rst
population study on asthma control in
Portugal, no previous data is available for
comparison.
1514
Migration and asthma medication in
international adoptees and immigrant
families in Sweden
Braback, L1; Vogt, H2; Hjern, A3
1
Umea University, Occupational and Environmental
Medicine, Umea, Sweden; 2Division of Pediatrics,
Linkoping University, Linkoping, Sweden; 3Karolinska
Institute, Stockholm University, Centre for Health
Equity Studies, Stockholm, Sweden
Background: Studies of asthma in migrant
populations illustrate the effects of envi-
ronmental changes. In this register study
we investigated the importance of exposure
to a Western lifestyle in different phases of
development in Swedish residents with an
origin in regions in the world where
asthma usually is less prevalent.
Method: The study population comprised
24 252 international adoptees, 47 986 for-
eign-born and 40 971 Swedish-born with
foreign-born parents and 1 770 092 Swed-
ish-born residents with Swedish-born par-
ents (age 625 years). Purchased prescribed
inhaled corticosteroids (ICS) during 2006
were used as an indicator of asthma.
571

Result: International adoptees and children
born in Sweden by foreign-born parents
had three to four fold higher rates of
asthma medication compared with foreign-
born children. The odds ratios of asthma
medication declined persistently with age
at immigration. For adoptees the odds
ratios (ORs) compared with infant adop-
tees were 0.78 (95% condence interval
(CI) 0.710.85) for those adopted at 1
2 years, 0.51 (0.420.61) at 34 years and
0.35 (0.270.44) after 5 or more years of
age. Corresponding ORs for foreign-born
children with foreign-born parents immi-
grating at 04 years, at 59 years, at 10
14 years and at 15 years or more were 0.73
(0.630.86), 0.56 (0.460.68) and 0.35
(0.280.43), respectively. The odds ratios
were only marginally affected by adjust-
ment for region of birth and socioeco-
nomic indicators.
Conclusion: Age at immigration is a more
important determinant of purchased ICS
than population of origin. This indicates
the importance of environmental factors
for asthma in schoolchildren and young
adults.
1515
Urbanisation is associated with an
increased prevalence of childhood
asthma in diverse, small rural communi-
ties in Latin America
Rodriguez, A1; Barreto, M2; Rodrigues, L3; Cooper, P1
1
Centro de Investigaciones FEPIS, Quininde, Ecuador;
2
Instituto de Saude Coletiva, Salvador de Bahia, Brazil;
3
Infectious Diseases Epidemiology, London School of
Hygiene and Tropical Medicine, London, United
Kingdom
Background: Studies conducted in transi-
tional communities from Africa and Asia
have pointed out the processes of urbaniza-
tion and modernization as the possible
responsible for the asthma increase in
developing regions. In Latin America there
is limited evidence for the impact of urban-
ization on asthma prevalence, especially in
transitional communities. The aim of the
present study was to explore how the pro-
cess of urbanization may explain differ-
ences in prevalence of childhood asthma
between diverse, small rural communities
in a tropical region of Ecuador.
Method: An ecological study was con-
ducted in 59 communities using several
indicators of urbanization classied in
three different groups (socioeconomic, life-
style and urban infrastructure). Categorical
Principal Component Analysis was used to
develop 4 indexes: one for each group of
indicators and a summary urbanization
index which included the most representa-
tive variables in each group. Spearmans
correlation was used to explore the rela-
572
Poster Session 64 Allergy and asthma: worldwide experiences, risk factors and epidemiology
tionship between each index and commu-
nity asthma prevalence. We also analyzed
the association between each constituent
variable for each of infrastructure, socio-
economic, and lifestyle indicator groups
and community asthma prevalence using
bivariate and multivariate linear regression,
weighted for the inverse of the variance of
the mean of community sizes.
Results: The overall prevalence of asthma
was 10.1% (range 0.031.4% between
communities).Three of the 4 indexes pre-
sented signicant associations with com-
munity asthma prevalence: socioeconomic
(r = 0.295, P = 0.023), lifestyle (r =
0.342, P = 0.008) and summary urbani-
zation index (r = 0.355, P = 0.006).
There was only a weak association
between the urban infrastructure index
and asthma prevalence (r = 0.173, P =
0.190). Indicators related with a better
socioeconomic status of the communities
and a larger presence of urban lifestyle
was associated with greater asthma preva-
lence.
Conclusions: Our data provide evidence
that the prevalence of asthma in transi-
tional communities increases with increas-
ing levels of urbanization, particularly
when measured using factors associated
with a greater socioeconomic level and
changes in lifestyle.
1516
Paradox in the prevalence of allergic dis-
orders: high prevalence of allergic rhinitis
and low prevalence of asthma in the
Khorassan province, Iran
1
Kazemi Azghadi, S ; Noorani Hasan Kiadeh, S ;1
Fereidouni, F1; Fereidouni, M1; Varasteh, A2
1
Birjand University of Medical Sciences, Birjand,
Islamic Republic of Iran; 2Mashhad University of
Medical Sciences, BuAli Research Institute, Mashhadi,
Islamic Republic of Iran
Background: Allergic disorders including
asthma, allergic rhinitis and atopic derma-
titis are a common health problem world-
wide and it has been reported that the
prevalence is increasing throughout the
world although the prevalence and the rate
of increase in not similar in all parts of the
world. Studies conrmed the existence of
differences, within and between countries,
in the prevalence of asthma, rhinitis and
eczema. Differences in methodology and
diagnostic criteria for measuring the inci-
dence could account for some disparities,
but they cannot explain all of them. The
aim of this study was to evaluate the prev-
alence of allergic disorders and related
symptoms in two cities in Khorassan prov-
ince of Iran, Mashhad and Boshrooyeh,
which are quite different in geo-climatic,
socio-economic and environmental factors.
Allergy 66, Suppl. 94 (2011): 482642  2011 John Wiley & Sons A/S
Method: A translated version of ECRHS
questionnaire was distributed to a random
selected population in Mashhad and Bosh-
rooyeh. Criteria for denition of asthma
and allergic disorders were based on the
presence of allergic related symptoms or
diagnosis of disease by physician. To com-
pare the result with ISAAC surveys, the
prevalence of allergic diseases was measured
for1314 years old responders as well.
Result: In total, 2364 subjects participated
in the survey (response rate 82%, mean
age = 26, M/F ratio 1/1.13). In both cities
the prevalence of self reported documented
asthma and asthma dened based on
asthma related symptoms found very low
(from 0.2% to 2.8%) and there was no sig-
nicant difference between two cities. The
prevalence of allergic rhinitis was signi-
cantly higher in Mashhad than Boshrooyeh
(22% versus 15.5% respectively,
P < 0.05). The prevalence of asthma and
allergic rhinitis in 1314 years old people
was 1.1% and 14.8% respectively.
Conclusion: Our study shows high preva-
lence of allergic rhinitis and very low prev-
alence of asthma in both cities. The results
suggest that, at least in some cases, asthma
is not necessarily directly related to other
atopic disorders and the underlying factors
for the low prevalence of asthma should be
dened in the future.
1517
Prevalence of asthma, allergic rhinitis
and eczema in the elementary school
(ISAAC study) in Iran
Ghaffari, J1; Mohammadzadeh, I2; Khalilian, A2; Saffar,
M2; Mohammadjafar, H2; Davoudi, A2
1
Mazandaran university of medical sciences, Allergy
and immunology, Sari, Islamic Republic of Iran;
2
Islamic Republic of Iran
Objective: Allergic diseases including
asthma, Allergic rhinitis (AR) and eczema
are common chronic disease in childhood.
Aim of this study to explore the prevalence
of asthma, AR and eczema.
Methods: This is a descriptive cross-sec-
tional study. This study was conducted on
the questionnaire according the ISAAC
protocol. Each elementary schools child
(712 years old) was a cluster. This study
was done in Sari, the capital of Mazanda-
ran province in the north of Iran.
Findings: Out of 1818 students; 646 cases
were allergic disorder (35%) including; 223
(12%) were asthma; 318 (17%) AR and
105 (6%) were eczema. Total allergic boys
were higher (391-65%) than girls (255-
40%) (P < 0.05).
Conclusion: In our study, the prevalence of
asthma, AR and eczema are higher in boys
than in girls, they are in a moderate preva-
lence compare to other countries.
Poster Session 64 Allergy and asthma: worldwide experiences, risk factors and epidemiology
1518
Associations between asthma and bron-
chial hyper-responsiveness with allergy
and atopy phenotypes in urban black
South African teenagers
Levin, M1; Muloiwa, R2; Motala, C1
1
Division of Allergy, University of Cape Town, Cape
Town, South Africa; 2Division of Ambulatory and
Emergency Paediatrics, University of Cape Town, Cape
Town, South Africa
Background: Epidemiological studies in
South Africa show increasing prevalence
rates of asthma and allergic sensitisation in
both urban and rural Black African com-
munities, and narrowing of the urban-rural
gradient. There is a paucity of current data
on bronchial hyper-responsiveness (BHR)
in urban Black African children, associa-
tions between asthma and BHR and the
relationship between BHR, allergen sensiti-
sation and other atopic diseases.
Objectives: To determine asthma and
allergy phenotypes in unselected urban
Black African (Xhosa-speaking) teenagers
and to associate BHR with asthma, other
atopic diseases and allergen sensitisation.
Method: Cross sectional study of two hun-
dred and eleven urban high school Xhosa
children. Modied ISAAC questionnaires
regarding asthma, eczema and rhinitis were
administered. BHR was assessed by meth-
acholine challenge using hand-held nebulis-
ers. Skinprick tests (SPT) were performed
to 8 aeroallergens and 4 food allergens.
Result: Asthma was reported in 9% and
16 % demonstrated BHR. Rhinitis was
reported in 48% and eczema in 19%.
Asthma was strongly associated with BHR.
Asthma was associated with eczema
whereas BHR was associated with rhinitis.
SPTs were positive in 34% of subjects; ae-
roallergens in 32%, food allergens 5%.
The most common sensitivities were to
house dust mites (HDM) and German
cockroach. BHR was associated with sensi-
tivity to any aeroallergen, Cat, HDM,
Cockroach and Bermuda grass. Number of
positive SPTs was associated with asthma
and BHR. With each level of SPT positiv-
ity there is 40% increased prevalence of
asthma and 70% increased prevalence of
BHR. The rate of allergen sensitisation in
subjects with BHR (72%) was much higher
than those without BHR (28%); house
dust mite sensitivity: 69% in subjects with
BHR and 18% in those without BHR.
Conclusion: These are the highest rates of
allergen sensitisation in subjects with BHR
documented in the African setting and the
widest difference in sensitisation rates
between subjects with and without BHR.
Allergy 66, Suppl. 94 (2011): 482642  2011 John Wiley & Sons A/S
1519
tropicalis, was 60.2% in the case group
Prevalence of asthma, allergic rhinitis (asthma in previous 12 months) and 40%
and other allergic conditions in in control group (no symptoms of asthma,
Colombia, 2009 AR, or AD in the last year plus no previ-
ous medical diagnosis of any of the three
Dennis, R1; Caraballo, L2; Garcia, E3; Rojas, M4;
Aristizabal, G5; Rondon, M4; Perez, A6; Penaranda, A7;
conditions).
Barragan, A1; Ahumada, V2; Jimenez, S2 Conclusion: This study shows that the cur-
1
Internal Medicine and Investigation Departments, rent prevalence of symptoms of asthma,
Fundacion Cardioinfantil-Instituto de Cardiologia,
allergic rhinitis, and atopic dermatitis in
Bogota, Colombia; 2Institute for Immunological
Research, University of Cartagena, Cartagena, six cities of Colombia is high, being disease
Colombia; 3Pediatric Allergy Department, Fundacion severity very frequent. Atopy is highly
Santa Fe de Bogota, Bogota, Colombia; 4Department of
associated with all three conditions.
Clinical Epidemiology and Biostatiscs, Faculty of
Medicine, Pontificia Universidad Javeriana, Bogota,
Colombia; 5Pediatric Department, Universidad El
Bosque, Bogota, Colombia; 6Division of Biostatistics,
School of Public Health, The University of Texas Health 1520
Science Center at Houston, Austin, United States; Prevalence and characterisation of
7
Fundacion Santa Fe de Bogota, Otorhinolaryngology
asthma and allergic diseases in adoles-
Department, Bogota, Colombia
cents of Belem (Amazonic Region), Brazil
Background: Studies estimating the preva- Sole, D; Barreto, B
lence of asthma and other diseases with an Department of Pediatrics, Sao Paulo Federal University,
Sao Paulo, Brazil
allergic background, such as rhinitis and
dermatitis, showed an important increase
Background: The prevalence of asthma
in most countries where such studies were
and other allergic diseases in Amazonic
conducted. In Colombia, a previous study
Region of Brazil is unknown. The preva-
in six cities showed that the prevalence, the
lence of asthma and possible associated
severity and the social consequences were
risk factors, as well as the prevalence of
worrisome. Ten years later, the surveillance
other allergic diseases in adolescents from
of these disease prevalent is warrant.
13 to 14 years old, residents in Belem, a
Method: An observational, cross-sectional
city of this region, was determined.
survey in six Colombian cities: Barranquil-
Method: A cross-sectional epidemiological
la, Bogota, Bucaramanga, Cali, Medellin,
study evaluating 3 725 adolescents from
and San Andres Island in Colombia was
Belem, who completed the standard
conducted. Subjects between 517 years
ISAAC-WQ (International Study of
old, were sampled in a school-based design
Asthma and Allergies in Childhood - Writ-
and subjects between 14 years old and
ten Questionnaire), following a case con-
1859 years old were selected using a com-
trol study on random sample of these
munity-based strategy. Current and cumu-
adolescents (126 asthmatics and 254 non-
lative prevalence of Asthma, Allergic
asthmatics) was performed. They were
Rhinitis (AR) and Atopic Dermatitis (AD)
assessed for possible risk factors by com-
symptoms were estimated overall and by
plementary questionnaire consisting of 33
city. We collected blood samples to deter-
questions relating to personal data, family
mine levels of allergen-specic IgE anti-
and environmental, answered by parents
body and total IgE. Parental consent was
and assessed skin prick tests for evaluation
obtained for children, teen-agers also pro-
of atopic sensitization. The association
vided written assent.
between asthma and risk factors was evalu-
Result: A total of 5 978 subjects were
ated by logistic regression analysis with
included. Overall current prevalence of
statistical signicance level of 5%.
asthma was 12.1% [95% CI: 10.513.6%],
Result: 3 708 adolescents who answered to
and varied among cities [11.3% to 14.5%].
the ISAAC-WQ appropriately were
Indicators of asthma severity in the past
enrolled, and 1 782 (48%) were male. The
12 months: night awakenings due to symp-
prevalence of asthma, rhinitis and atopic
toms 58% [95% CI: 50.365.2%] and visits
eczema in the last 12 months were 20.7%,
to an emergency room or hospitalization
44.2% and 12.5% respectively. The medi-
43% [95% CI: 36.349.2%]. Overall preva-
cal diagnosis of asthma, rhinitis and atopic
lence of current AR symptoms was 32%
eczema was 29.3%, 30.7% and 10.4%
[95% CI: 29.533.9%], similar by sex but
respectively. Risk factors signicantly asso-
varied across cities (28.7% to 39.5%) and
ciated with asthma were: presence of rhini-
by age group being greater in children
tis diagnosed by physician (OR = 1.7; CI
from 517 years 46.9% [95% CI: 40
95%:1.22.9), current rhinitis (OR = 3.2;
53.9%]. Overall, prevalence of current AD
CI 95%:1.85.9), diagnosis of measles
symptoms was 21% [95% CI: 18.822.6%]
(OR = 4.7; CI95%:2.39.8), previous diag-
not signicant variation was noted among
nosis of tuberculosis (OR = 38.9; CI95%:
cities or by age group. The prevalence of
4.6328.0), high level of intradomiciliar
atopy, as dened by total IgE and specic
humidity (OR = 1.8; CI95%:1.13.2),
IgE levels against D. pteronyssimus and B.
573
 Poster Session 64 Allergy and asthma: worldwide experiences, risk factors and epidemiology

exposure to smokers (OR = 2.4; CI
95%:1.24.5) and breastfeeding (OR =
4.2; CI 95%:1.115.2). For atopic sensitiza-
tion assessment, the positivity for at least
one allergen was signicantly higher in
asthmatics adolescents (86.5% versus
32.4%; P < 0.0001).
Conclusion: The prevalence of asthma and
atopic eczema in adolescents in the city of
Belem was similar to the Brazilians aver-
age, but allergic rhinitis was signicantly
higher. Physician-diagnosed asthma was
more frequent than the presence of symp-
toms suggestive of asthma. Infectious,
nutritional and environmental factors, as
well the concomitant presence of allergic
rhinitis, were the main risk factors associ-
ated with the development of asthma in
these adolescents.

574 Allergy 66, Suppl. 94 (2011): 482642  2011 John Wiley & Sons A/S

The many faces of asthma
1521
The most common causes of chronic
nonproductive cough
Arbutina, S; Pejkovska, S; Arsovski, Z; Goseva, Z;
Busljetic, K; Karkinski, D; Kaeva, B
Clinic of Pulmology and Allergology, Skopje,
Macedonia, FYROM
Background: Chronic nonproductive cough
is a kind of cough not accompanied by
expectoration of mucus. Chronic nonpro-
ductive cough is very often single symptom
that indicates presence of disease. Aim of
our study as to investigate the most com-
mon causes of chronic nonproductive
cough.
Method: We included 60 patients 24 men
and 36 women. All of them were non smo-
ker. They complain for presence of non
productive cough longer then 8 weeks. Fol-
lowing tests were obtained by indications:
Physical examination, blood laboratory
tests, lung function tests, bronchoprovoca-
tive methacholine test, skin prick test, chest
X ray, sinus X ray, CT scan of chest, echo-
cardiography, microbilogical tests, oesoph-
ageal pH monitoring and bronchoscopy.
Result: From the results of our study we
have found chronic nonproductive cough is
usually caused by sinusitits (28 pts.) cough
related asthma was found in 4 pts., GERD
syndrome in 8 pts. Presence of viral infec-
tion was found in 4 patients, pulmonary
sarcoidosis in 1 patient, neoplastic disorders
in 5 pts., congestive heart failure 6 pts., side
effects of drugs (ACE inhibitors) in 4 pts.
Conclusion: We can conclude that in prac-
tice we should pay special attention to the
exact etiology of unproductive cough
which can help us for precise diagnosis and
proper treatment of the basic disease.
1522
Fractional exhaled nitric oxide and its
correlation with biopsy results in chronic
cough patients
Votruba, J; CAP, P; Statsny, B
Nemocnice Na Homolce, Prague, Czech Republic
Background: Chronic non-productive cough
in non-smokers with negative thoracic
X-ray and normal spirometry is frequently
encountered problem in the out-patient
departments of specialists both pneumolo-
gists and allergologists. A lot of those
Allergy 66, Suppl. 94 (2011): 482642  2011 John Wiley & Sons A/S
Poster Session 65
patient present with already ongoing treat-
ment with inhaled steroids. This work
attempted to nd valid markers of inam-
mation in our group of patients.
Method: ICS naive non-smokers with neg-
ative X-ray of the chest, ACE-inhibitors
non-users with no signs of UACS have
been examined by bronchoscopy and
FENO (Fractional Exhaled Nitric Oxide).
Combination of these two methods aimed
to conrm or exclude presence of bronchial
inammation. Recommendation for treat-
ment with ICS can be more valid if based
on positive results. Very small number of
patients with eosinophilic type of bronchial
inammation on biopsy (3 from total no.
48) was found. In the group of those 3
patients with positive nding of eosino-
philia in mucosa regarding changes there
were also high values of FENO, whereas in
all other allergy negative patients the was a highly signicant correlation between
FENO values were low.
Conclusion: There was a low incidence of
eosinophilic inammation in our group of
chronic cough patients. Bronchoscopy and
FENO were in good agreement in ruling
out or in conrmation of eosinophilic
inammation. FENO appears to be a good
discriminator for ICS treatment decision in
our group of patients.
1523
Exhaled breath temperature is influenced
by both age and diagnosis of respiratory
disease in children
Kralimarkova, T1; DuBuske, L2; Dimitrov, V3; Popov, T3
1 Conclusion: EBT measurement with the X-
Clinic of Allergy & Asthma, Sofia Medical University,
Sofia, Bulgaria; 2Immunology Research Institute of New
England, Gardner, MA, United States; 3Clinic of Allergy
& Asthma, Sofia Medical University, Sofia, Bulgaria
Background: Measurement of exhaled
breath temperature (EBT) has been sug-
gested as surrogate biomarker of airway
inammation. This approach has been
enhanced recently by the introduction of a
portable user-friendly device having a high
degree of precision and reliability. Most
EBT studies to date have employed adult
subjects. The aim of our recent efforts was
to assess confounding inuences that need
to be considered when analyzing EBT in
children.
Method: EBT was evaluated in 78 children
(46 boys, age range: 3 to 17 years): 56
referred to the outpatient pediatric depart-
ment of the Clinic of Allergy & Asthma in
Soa and 22 recruited as healthy controls
from among the families of the personnel
of the clinic. Measurements were done by
means of a hand-held device (X-halo, Del-
medica Investments Ltd, Singapore) fol-
lowing informed consent by the childrens
parents / guardians. Children and their
accompanying persons were briey
instructed on how to use the device. Diag-
noses of the children were based on the
overall judgment of the consulting physi-
cian, not taking into consideration the sub-
sequent EBT values.
Result: All children, irrespective of their
age and gender, were able to achieve mean-
ingful values of their EBT. There was no
correlation between the EBT and the otic
temperature measured in parallel. There
EBT and age among the healthy controls
(R = 0.75; P < 0.001). This correlation
remained strong when correlating EBT and
age in all 78 children, but the R was
reduced to 0.43, (P < 0.001), possibly due
to respiratory pathology in 22 asthma and
allergic rhinitis patients including those
with pollenosis out of the pollen season (11
patients), adenoid vegetations (7 patients),
viral upper airway infections (7 patients),
chronic cough (6 patients), and skin aller-
gic diseases (3 patients). One-way ANOVA
analysis disclosed differences in EBT across
this wide diagnostic range. The widest
spread of EBT values was among the chil-
dren with chronic cough.
halo hand held device is a non-invasive,
easily applied method in the environment
of a routine outpatient pediatric practice.
Patient age needs to be considered when
interpreting the results. More data is cur-
rently being obtained to allow insight into
the clinical relevance of EBT in various
disease states including asthma.
575
 Poster Session 65 The many faces of asthma

1524
Role of NOS2A variants in the associa-
tion between nitrite/nitrate in exhaled
breath condensate and response to skin-
prick test
Rava, M1; Bouzigon, E2; Varraso, R3; Huyvaert, H4;
Bousquet, J3; Le Moual, N3; Pin, I5; Demenais, F2;
Matran, R4; Nadif, R3
1
Inserm, CESP U1018, Paris, France; 2Inserm, U 946,
Paris, France; 3Inserm, CESP U1018, Villejuif, Paris,
France; 4Universite Lille Nord de France, Lille, France;
5
Inserm, U 823, Grenoble, France
Background: The exhaled fraction of nitric
oxide (FeNO) is known as a biomarker of
pulmonary inammation. Epidemiological
studies led to conicting results regarding
associations between FeNO, atopy and
asthma. Nitrite (NO2)) and nitrate (NO3))
are other indicators of NO synthesis. Total
NO2-NO3 we are exposed to derive from
endogenous and dietary sources (vegeta-
bles, cured meat). Nitric Oxide Synthase
(NOS) is the key enzyme in the formation
of NO. This study is aimed at evaluating
the associations between (1) NO2-NO3 in
exhaled breath condensate (EBC) and
response to Skin Prick Tests (SPT) (2) sin-
gle nucleotide polymorphisms (SNPs) in
NOS2A with NO2-NO3 and response to
SPT and (3) whether genetic factors mod-
ify the association between NO2-NO3 level
and response to SPT among adults from
the French Epidemiological case-control
family-based study on Genetics and Envi-
ronment of Asthma (EGEA).
Method: Positivity to SPT (SPT+) was
dened as a positive response (>=3mm
diameter) to at least 1 of 11 aeroallergens.
Total NO2-NO3 (converted nitrate plus
nitrite) level was measured in EBC. 37
SNPs belonging to NOS2A were consid-
ered. Odds ratios for SPT+ were estimated
with generalized estimating equations
(GEE) for logistic regression models
accounting for age and gender. Association
between the 37 SNPs with SPT+ and
NO2-NO3, interactions between NO2-NO3
and the SNPs on SPT+ were tested under
an additive genetic model.
Result: Data were available for 1327 adults
(43 years, 51% women, 37% asthmatics).
EBC NO2-NO3 level was unrelated to
asthma, diet, smoking habits. A positive
and borderline statistically signicant asso-
ciation was found between level of total
NO2-NO3 in EBC and SPT+ (adj OR =
1.38 (95% CI 0.99, 1.91), P = 0.06) and a
stronger association was found among
non-asthmatics (adj OR = 1.51 (95% CI
1.02, 2.22), P = 0.04) versus asthmatics.
No association was found between SNPs
in NOS2A and SPT. One SNP (rs4795067)
was signicantly associated with NO2-NO3
(b = 0.053, P = 0.046). Statistically signif-
icant interactions were found between ve
SNPs and EBC NO2-NO3 on SPT+, one
of which (rs11080358) remained statisti-
cally signicant after correction for the
effect of multiple testing. This result was
conrmed after adjustment for gender and
age.
Conclusion: Preliminary results show an
association between total NO2-NO3 and
SPT+ and suggest that this association
might be modulated by the genes that
encode the inducible isoform iNOS.
1525
Inflammatory markers and acid-base
equilibrium in exhaled breath condensate
of stable and unstable asthma patients
Tomasiak-Lozowska, M1; Zietkowski, Z1; Skiepko, R1;
Tomasiak, M2; Przeslaw, K2; Bodzenta-Lukaszyk, A1
1
Department of Allergology and Internal Medicine,
Medical University of Bialystok, Poland, Bialystok,
Poland; 2Department of Physical Chemistry, Medical
University of Bialystok, Bialystok, Poland
Background: Oxidative and acid stress play
an important role in the pathogenesis of
asthma. The aims of the study were to
assess markers of oxidative and acid stress
(pH, nitrite/nitrate, ammonia) in exhaled
breath condensate (EBC) in asthmatics and
to determine the potential relationship
between these parameters and asthma
severity, lung function and other indices of
airway inammation (exhaled nitric oxide -
FENO, total IgE, eosinophil cationic pro-
tein - ECP, blood eosinophilia).
Method: pH, ammonia and nitrite/nitrate
levels were measured in EBC samples col-
lected from 91 allergic asthmatics (22 with
stable, steroid-naive asthma, 35 with stable,
ICS-treated asthma and 34 with ICS-trea-
ted, unstable asthma) and 19 healthy vol-
unteers. The possible correlations of these
measurements with the results of other
tests used in the diagnosis of asthma as
well as laboratory tests commonly associ-
ated with asthma were also investigated.
Result: Both pH and ammonia values in
EBC were signicantly lower in asthmatics
than in healthy subjects; the lowest levels
of these two parameters were observed in
patients with unstable asthma. In the
group of stable, ICS-treated asthmatics,
EBC pH was signicantly higher than in
patients with unstable, ICS treated asthma.
We revealed statistically signicant lower
concentrations of ammonia in the group of
ICS-treated patients with unstable asthma
compared with those with stable asthma
and steroid-nave patients. The levels of
nitrites/nitrates in EBC were signicantly
higher in all groups of asthmatics than in
healthy volunteers; the highest concentra-
tions were observed in the group of
patients with unstable asthma. We revealed
statistically signicant positive correlation
between pH of EBC and ammonia levels in
EBC and negative correlation between
EBC pH and nitrites/nitrates or FENO or
blood eosinophil count in three groups of
asthmatics.
Conclusion: There is a relationship
between acid-base equilibrium and mark-
ers of oxidative stress in exhaled breath
condensate and other recognized parame-
ters of inammatory status in asthmatic
patients. All three parameters (pH, ammo-
nia, nitrite/nitrate) measured together in
EBC in asthmatics could be useful in the
assessment of airway inammation and
asthma severity.

Table 1. For abstract 1524.

Heterozygous
Adjusted for the minor Homozygous for
interaction allele (major the minor allele
Interaction Estimate estimate Adjusted allele as reference (major allele as
and 95% CI P-value and 95% CI P-value category) reference category)
(#signicant
Minor after correction (additive model, (adj
allele for multiple adj for age, for age,
SNP Alleles frequency (additive model) testing) gender) gender) N OR (95% CI) N OR (95% CI)
rs10853181 A/G 0.40 0.47 (0.09, 0.84) 0.014 0.39 ()0.01, 0.80) 0.059 530 0.99 (0.70, 1.40) 196 1.68 (1.05, 2.68)
rs11080358 G/A 0.15 )0.95 ()1.52, )0.37) 0.001# )0.86 ()1.48, )0.24) 0.007 303 1.10 (0.76, 1.59) 29 1.91 (0.73, 4.98)
rs2531894 G/A 0.40 )0.44 ()0.81, )0.06) 0.022 )0.34 ()0.75, 0.06) 0.094 536 1.00 (0.71, 1.42) 195 1.70 (1.06, 2.70)
rs2779248 T/C 0.40 )0.46 ()0.87, )0.05) 0.028 )0.52 ()0.97, )0.08) 0.020 360 1.03 (0.74, 1.44) 568 1.28 (0.78, 2.08)
rs34920947 G/C 0.40 )0.43 ()0.80, )0.05) 0.026 )0.33 ()0.74, 0.07) 0.106 536 0.99 (0.70, 1.40) 195 1.72 (1.07, 2.75)

576 Allergy 66, Suppl. 94 (2011): 482642  2011 John Wiley & Sons A/S
Poster Session 65 The many faces of asthma
1526
Effect of different periods of argon de-
aeration on exhaled breath condensate
pH
Barato, D1; Orosa, B2; Prieto, L3; Marin, J3
1 1
Doctor Peset University Hospital, Allergy, Valencia,
Spain; 2Doctor Peset University Hospital, Pneumology,
Valencia, Spain; 3Universidad de Valencia, Medicina,
Valencia, Spain
Background: The pH of exhaled breath
condensate (EBC) immediately tested tends
to be unstable and therefore, to enhance
the stability of the determinations, deaera-
tion with argon can be performed. Little
information is available, however, on the
duration of argon deaeration required to
achieve a stable pH in EBC samples. The
aim of this study was to identify differ-
ences in EBC pH after argon de-aeration
for 2, 4 and 8 min.
Method: EBC pH was determined in EBC
samples from 48 subjects with allergic rhi-
nitis (11 with associated asthma) and 14
healthy volunteers. EBC was collected
using the RTube collection system (Respi-
ratory Research, Inc, Charlottesville, VA)
during tidal breathing for 10 min without
a nose-clip. Each EBC sample was divided
into three aliquots and the pH without
deaeration was measured. The three aliqu-
ots were then de-aerated (bubbling argon)
for 2, 4 and 8 min prior to pH measure-
ment.
Result: The mean (95% CI) pH values
obtained from samples analysed after
4 min [7.66 (7.527.80)] and 8 min [7.70
(7.557.85)] of argon de-aeration were sig-
nicantly less acidic (P < 0.001) than
those identied after 2 min of deaeration
[7.53 (7.407.66)]; differences between pH
values at 4 and 8 min were not signicant.
Compared with nondeaerated samples, the
mean (95% CI) pH values increased in
1.11 (1.031.19), 1.22 (1.131.30) and 1.24
(1.151.33) after 2, 4 and 8 min of deaera-
tion. Changes in pH after 4 and 8 min of
deaeration were signicantly greater than
those after 2 min, the mean difference
being 0.11 (95% CI, 0.020.20, P < 0.05)
and 0.13 (95% CI, 0.040.22, P < 0.01),
respectively. Differences between changes
at 4 and 8 min were not signicant.
Conclusion: The magnitude of change in
EBC pH after argon deaeration is approxi-
mately one, and stabilization of this change
is achieved after argon deaeration for
4 min. Therefore, a deaeration period of
4 min is recommended to stabilise the pH
of EBC samples.
Allergy 66, Suppl. 94 (2011): 482642  2011 John Wiley & Sons A/S
1527 1528
A comparison of two methods to calcu- Exhaled nitric oxide and asthma control
late peripheral nitric oxide exchange in measured by clinical score and exercise-
the lungs induced bronchoconstriction
Prieto, L1; Barato, D2; Marin, J1 Bosque-Garcia, M1; Asensio-de la Cruz, O2; Jaramillo-
Universidad de Valencia, Medicina, Valencia, Spain; Hidalgo, D3; Valdesoiro-Navarrete, L2; Costa-Colomer,
2
Hospital Dr Peset, Allergy, Valencia, Spain J1; Penas-Aguilera, A1; Garcia-Gonzalez, M1
1
Hospital de Sabadell. Universitat Autonoma de
Barcelona, Pediatric Allergy, Sabadell, Spain; 2Hospital
Background: Two methods have been de Sabadell. Universitat de Barcelona, Pediatric Allergy,
developed to account for the impact of air- Sabadell, Spain; 3CAP Badia del Valles, Primary Care,
way nitric oxide (NO) production on alve- Badia del Valles, Spain
olar NO concentration. In the rst method
Background: Nitric oxide (NO) plays an
(Condorelli et al. JAP. 2007; 102:417425),
important role in several biological pro-
small airway/alveolar NO (CANO) is cor-
cesses as inammation. Exhaled NO (eNO)
rected using large airway NO (JawNO)
is increased in asthma. 40-90% of asthma
value, whereas the second method (Kerckx
patients develop exercise-induced broncho-
et al. JAP. 2008; 104: 918924) requires
constriction (EIB). eNO can be correlated
knowledge of the exhaled NO concentra-
with forced expiratory volume in 1 second
tion at a ow of 50 ml/s (FENO50). The
(FEV1) decrease after exercise. As studies
aim of the present study was to determine
have shown different evidence about this
the inuence of the correction method on
correlation, our aim was to determine if
CANO values.
eNO assessed in daily clinical practice
Method: A total of 91 adult subjects (27
tracks bronchial hyperreactivity. Determin-
asthmatics, 46 patients with allergic rhinitis
ing whether eNO predicts EIB, after Free
and 18 healthy volunteers) were studied.
Running Test (FRT), measured by spirom-
FENO50 level as well as exhaled NO at
etry; assessing correlation between eNO
expiratory ow rates of 100, 200 and
and asthma control perception, measured
300 ml/s were measured using a chemilumi-
by the Asthma Control Test (ACT) Score.
nescence analyzer (NiOx; Aerocrine, Solna,
Method: Prospective observational design
Sweden). The technique of Tsoukias and
including 30 children, previously diagnosed
George (JAP. 1998; 85: 653666) was used
and followed up, with diagnose of allergic
to calculate JawNO and CANO values.
asthma (aeroallergen IgE-mediated sensiti-
CANO was adjusted for the trumpet shape
zation), consecutively visited in the Allergy
of the airway tree and axial diffusion from
and Respiratory Department of Sabadell
central to peripheral airways using both
Hospital. The following tests were used:
the Condorelli and Kerckx methods.
ACT, basal eNO and spirometry, 8 min
Result: The mean (95% CI) CANO values
FRT, post-exercise eNO and spirometry,
obtained with the Condorelli method were
10 and 20 minutes post-exercise spirome-
1.27 ppb (0.931.60), compared to 0.87
try. All tests were performed using Dato-
ppb (0.551.19, P < 0.001) with the Ker-
spir 600 (Sibelmed), NO Vario Analyser
ckx method. Differences in CANO values
(v4.14). EIB was dened as a decrease of
obtained with each method were identied
FEV1 higher than 12%. We considered an
in both subjects (n = 55) with increased
appropriate FRT when 80% of age-
FENO50 values (> 25 ppb) and in those
adjusted maximum heart rate was
(n = 36) with normal ( 25 ppb) FENO50
achieved. Statistic analysis performed with
values, but were signicantly greater in the
SPSS 15.0 version.
group with increased FENO50 values
Result: Basal eNO levels (37.8 32 ppb)
(P = 0.01). Furthermore, differences
were signicantly higher than post-exercise
between the two methods were not totally
eNO levels (28.1 23.7 ppb) (paired t-
consistent and were more evident in sub-
test = 5.19, fd = 27, P-value <0.001).
jects with CANO values >1 ppb.
FEV1 signicantly decreased after FRT
Conclusion: This study demonstrates signif-
(5.1 5.4%) (paired t-test = 5.21,
icant differences in CANO values obtained
fd = 29, P-value <0.001). ACT score
with the two methods reported to adjust
(median 24, range 1925) showed this
for the trumpet shape of the airway tree
group of patients good-control self percep-
and axial diffusion from central to periph-
tion. Basal eNO correlated with post-exer-
eral airways. Differences between the two
cise eNO (R2 = 0.965, P < 0.001).
methods are not totally consistent and are
Decrease in eNO levels correlated with the
signicantly greater in subjects with
decrease of FEV1 (R2 = 0.225,
increased FENO50 values. These results sug-
gest that the CANO levels obtained with P = 0.011). We did not nd correlation
each approach are not equivalent and can- between basal eNO and ACT score
not be used interchangeably. (R2 = 0.033, P > 0.05), basal eNO and
FEV1 decrease (R2 = 0.169, P > 0.05).
The analysis of the subgroup of high bron-
chial hyperresponsiveness (FEV1 decrease
577

>10%) (n = 7), revealed a correlation
between basal eNO and FEV1 decline
(R2 = 0.88, P = 0.02), but not between
basal eNO and ACT (R2 = 0.08,
P > 0.05), neither decreased eNO.
Conclusion: In this study eNO did not
show utility as a predictor of EIB in a clin-
ically-stable group of asthma patients.
Among the group of highest bronchial
hyperresponsiveness eNO production is sig-
nicantly increased.
1529
Relationship between skin prick test,
peripheral eosinophil counts, serum total
& specific IgE and severity in atopic
asthma
Mahmoud, H1; Elqady, M2; Mohamed, H2
1
Chest Department, Sohag University Hospital, Sohag -
Elmaragha-Nagecc Alhakim, Egypt; 2Clinical Pathology
Department, Sohag University Hospital, Sohag, Egypt
Background: Atopic sensitization is a well-
known risk factor for asthma and bron-
chial hyperresponsivness. IgE-mediated
responses contribute to allergy and asthma.
Blood eosinophilis are known to be an
indirect marker of airway inammation in
asthma.
Objective: To evaluate the interrelationship
between skin prick test, serum total & spe-
cic IgE levels, eosinophilia and asthma
severity in atopic asthmatics.
Method: One hundred ten atopic asthmatics
patients were randomly selected and 30
matched control subjects were included. All
were submitted to detailed clinical history
and examination , pulmonary function test-
ing skin prick testing to 17 common aller-
gens, blood eosinophil counts and both
total and specic IgE levels. Asthma severity
was assessed according to GINA guidelines.
Result: There was a highly signicant rela-
tionship between total IgE, eosinophil
counts & percent (P < 0.0001) from one
hand and severity of asthma from the other
hand. There was a signicant increase in
severity of asthma with the increasing num-
ber of positive allergens in SPT & also
when asthma was associated with other
allergic diseases (P < 0.04). Increased
number of positive allergens was associated
signicant increase in the level of total IgE,
eosinophil count & percent (P < 0.0002).
Total IgE was signicantly higher in atopic
asthmatics with eosinophilia than those
without (P < 0.0001). Eosinophil count &
percent, total IgE and specic IgE were sig-
nicantly higher in patients than controls
(P < 0.0001). The most common allergens
for atopic asthma in our locality were D.
pternoyssinus, followed by D. farinae then
Cockroach extract.
Conclusion: Atopic asthmatic patients with
a higher serum concentration of both total
578
Poster Session 65 The many faces of asthma
IgE and specic IgE, blood eosinophilia agement and regularly follow-up and moni-
and sensitivity to more than one allergens toring. The work has to take place patient
in SPT have a higher asthma severity. safe and with careful documentation. The
quality of care has to be developed system-
atically and continuous, and secured and
prosecute in accordance with medical sci-
1530
ence and well-tried experience.
Job description for registered nurses
specialised in asthma/allergy chronic
obstructive pulmonary diseases in Swe-
den 1531
Zakrisson, A1; Jonsson, M2; Jagorstrand, B3 Skin reactivity to autologous plasma,
1
Orebro University, Family Medicine Research Centre, coagulation markers, and vascular endo-
Orebro, Sweden; 2Astrid Lindgrens Hospital, Karolinska thelial growth factor in patients with
University Hospital, Childrens Outpatient Clinic,
non-allergic asthma
Stockholm, Sweden; 3Allergy Competence Centre
South, Lund, Education Coordinator, Lund, Sweden Asero, R1; Tedeschi, A2; Cugno, M3
1
Clinica San Carlo, Ambulatorio di Allergologia,
Background: The nursing of patients with Paderno Dugnano, Italy; 2Fondazione IRCCS Ca
Granda, Ospedale Maggiore Policlinico, U.O.
asthma, allergy and Chronic Obstructive Allergologia e Immunologia Clinica, Milano, Italy;
Pulmonary Disease (COPD) differ a lot in 3
Universita di Milano e Fondazione IRCCS Ca Granda,
Sweden and how much of the nurses care Ospedale Maggiore Policlinico, Dipartimento di
Medicina Interna, Milano, Italy
and competence these patients receive.
Studies, both nationally and internation-
Background: Non-allergic asthma patients
ally, have shown that patients with asthma
show frequently autoreactivity as do
and COPD diseases are not optimally
subjects with chronic urticaria (CU).
controlled. A requirement for effective
Activation of the coagulation cascade and
recourse utilization in asthma, COPD and
hyper-expression of VEGF were recently
allergy care is development of care
found in CU, and there is some indication
chains with structural workload and rou-
that the same may occur in asthma. We
tines for information and education to
investigated autoreactivity, activation of
patients and their relatives. The facts that
the coagulation cascade, and expression of
allergic diseases have become more com-
VEGF in patients with non-allergic asthma.
mon have resulted in new demands on
Methods: Twenty-one adults with non-
health professionals competence and
allergic asthma underwent autologous
efforts. Although there are some evidence
plasma skin test (APST) and the measure-
that the input of nurses work are relevant
ment of plasma levels of fragment F1+2,
in the treatment of asthma/allergy/COPD,
D-dimer, and VEGF. APST was carried out
still nurses work is unclearly dened in
in 10 subjects with allergic rhinitis, 5 with
clinical guidelines or care programs. The
allergic contact dermatitis and 3 with aller-
guidelines that exist describe the nurse
gic asthma as pathologic controls. Twenty-
working tasks in different degree. There-
one healthy subjects sex- and age-matched
fore a job description for the asthma/
with patients served as normal controls.
allergy/COPD-nurses was needed to be
Results: The APST scored positive in 19/
able to give good patient safe care all over
21 (90%) patients versus 0/21 controls.
the country.
Mean fragment F1+2 plasma levels were
Aim: To develop a job description for
signicantly higher in asthmatic patients
asthma/allergy and COPD nurses to provide
(267 +/-243 pmol/l) than in controls
patients with asthma, allergy, COPD an
(150 +/-51 pmol/l; P = 0.0001). Similarly,
improved and safe disposal of medical care.
plasma levels of both D-dimer and VEGF
Method: The working team has made an
were signicantly higher in patients than in
inventory of existing national and interna-
controls (D-dimer: 2364 +/-1467 versus
tional knowledge over what is important in
1301 +/-525 pmol/L; P = 0.0001; VEGF:
nursing care to be able to handle a patient
1721 +/-2566 versus 76 +/-375 fmol/l;
safe and patient focused care. This job
P = 0.0001). A trend toward increased
description will be used for all types of
levels of F1+2, D-dimer and VEGF was
clinics where the asthma, allergy and
found in patients with a more severe dis-
COPD nurse working.
ease according to GINA classication.
Result: The asthma/allergy/COPD-nurse
Conclusion: Non-allergic asthma is charac-
has specic competence domains in caring
terized by skin reactivity to autologous
science, medical science, pedagogic, co-oper-
plasma and presence of circulating vasoac-
ation in team and with other caring at all
tive factors related to blood coagulation
levels together with research and develop-
and endothelium. These ndings might be
ment. The objective in sight of knowledge
of help to clarify the pathogenesis or to
and the attitude in nursing care is health
detect novel therapeutic strategies for this
from the particular perspective which can be
disease.
reached by patient education to self-man-
Allergy 66, Suppl. 94 (2011): 482642  2011 John Wiley & Sons A/S
Poster Session 65 The many faces of asthma
1532
Comprehensive evaluation of coughing,
transforming growth factor-b, interleukin-
13, interleukin-8, and airway reversibility
can facilitate the differential diagnosis of
asthma
Souma, R; Watanabe, H; Wakayama, T; Ochiai, A;
Sagara, H
Dokkyo Medical University Koshigaya Hospital,
Respiratory Medicine, Koshigaya, Japan
Background: Asthma and other diseases
associated with allergic airway inamma-
tion have been suggested to be character-
ized by the local involvement of
eosinophils, T cells, and mast cells in the
airways. Recently, neutrophils have been
implicated in the development of severe
asthma. Allergic airway inammation is
thus characterized by inammatory cell
inltration and activation in association
with injury to the airway epithelium.
Repeated episodes of allergic inammation
are thought to induce airway remodeling,
causing irreversible expiratory ow limita-
tion and persistent airway hyperresponsive-
ness, progressing to chronic, more severe
disease. This inammatory cascade of
events interacts in a complex fashion with
genetic factors (multiple factors) and envi-
ronmental factors, leading to the develop-
ment and progression of asthma. In
particular, interleukin (IL)-4, IL-5, and IL-
13 produced by Th2 cells are considered to
play a central role in allergic airway
inammation. However, specic markers
for the diagnosis of asthma remain unclear.
At present, a diagnosis of asthma contin-
ues to be based on symptoms or laboratory
abnormalities such as eosinophilia. Retro-
spective studies performed by research
groups including our institution have
shown that coughing is the most important
initial symptom of asthma.
Method: In the present study, we prospec-
tively measured airway hyperresponsive-
ness, blood eosinophil counts, exhaled
nitric oxide concentrations, respiratory
function, and concentrations of various
cytokines in patients with a chief complaint
of cough who were not given a diagnosis
of asthma. Our main objective was to
determine whether these variables are
important factors in the differential diag-
nosis of asthma and cough-variant asthma.
Result: The results showed that asthma
was associated with a signicantly higher
frequency of airway reversibility, as well as
a trend towards lower 50% and 25%
forced vital capacity expiratory ow rates
on respiratory function testing. The
exhaled nitric oxide concentration was
higher in patients with asthma. As for
cytokines, concentrations of transforming
growth factor-b and IL-13 were high in
asthmatic patients, whereas the concentra-
Allergy 66, Suppl. 94 (2011): 482642  2011 John Wiley & Sons A/S
tion of IL-8 tended to be higher in non-
asthmatic patients.
Conclusion: These results suggested that a
comprehensive evaluation of symptoms,
airway reversibility, and the levels of vari-
ous cytokines can facilitate the differential
diagnosis of asthma and non-asthma.
1533
Asthma control in patients with adult
onset disease
Marco Martn, G1; Alvarez-Perea, A1; de Miguel Dez, J2;
Magan Tapia, P3; Sanz de Burgoa Gomez-Pinan, V4;
Pelta Fernandez, R1
1
Hospital General Universitario Gregorio Maranon,
Allergy Service, Madrid, Spain; 2Hospital General
Universitario Gregorio Maranon, Neumology Service,
Madrid, Spain; 3Hospital Universitario 12 de Octubre,
Investigation Unit, Epidemiologic Clinic, CIBER
Epidemiology and Public Health, Madrid, Spain; 4Pfizer
Pharmaceuticals, Medical Advisor, Madrid, Spain
Background: Our main objetive was to
evaluate the asthma control in those
patients who initiated asthma disease after
12 years old.
Method: An epidemiologic, multicentric,
transversal, observational, case-control
study was carried out. Patients with asthma
onset after 12 years old who required medi-
cal assistance in an Allergy or Pneumology
clinic for any reason were included. For all
cases, socio-demographic data, personal
and familiar reports, clinical features, respi-
ratory functions tests, diagnostic and treat-
ment were collected. Asthma control grade
was determined by ACT grade (Asthma
Control Test). In addition, for all cases,
quality of life questionnaires such as
SGRQ (St Georges Respiratory Question-
naire) and AQ20 (Airways Questionnaire
20) were included. Statistical analysis was
carried out with SAS Statistical Proce-
dures, version 8.2. For the main factors of
asthma control determination, a logistical
multivariate regression test was performed.
Result: 247 patients were evaluated (42.6%
male, 57.4% female) with an average age
of 28.3 8.2 years. 41.5% of these
patients presented mild persistent asthma,
31.9% intermittent asthma, and 25.8%
moderate persistent asthma. ACT average
score was 20.5 3.8. 30.9% of the cases
presented insufcient asthma control (ACT
19) and 69.1% sufcient asthma control
(ACT >19). It was detected that the main
factors inuencing asthma control were the
total punctuation of the SGRQ question-
naire (OR 0.941; IC 95% 0.9020.982) and
the total punctuation of the AQ20 ques-
tionnaire (OR 0.829; IC 95% 0.7190.956),
as observed in the logistical multivariate
regression test. None of the characteristics
related to patients proles were found sta-
tistically signicant among the analysis
groups.
Conclusion: In this study, we demonstrate
that the total punctuation of both ques-
tionnaires, SGRQ and AQ20 are the main
factors for asthma control assessment. A
one point increase in SGRQ and, conse-
quently, having a greater alteration of
quality of life, results in a 1.1 times
increase of the probability of having insuf-
cient asthma control. On the other hand,
a one point increase in AQ20 punctuation
and, hence, having a worse quality of life,
increases the probability of having insuf-
cient asthma control 1.2 times.
1534
Independent predictors influencing
Health Related Quality of Life (HRQoL) of
patients with asthma
Uchmanowicz, I1; Jankowska-Polanska, B2; Panaszek,
B3; Fal, A3
1
Faculty of Health Science, Medical University,
Wroclaw, Poland; 2Wroclaw, Medical University, Faculty
of Health Science, Wroclaw, Poland; 3Medical
University, Faculty of Health Science, Wroclaw, Poland
Background: The burden of asthma
includes a substantial impact on Health
Related Quality of Life (HRQoL). As for
most chronic diseases its also widely
accepted for asthma that monitoring of
social and economic impact of the disease
should include measures of its impact on
HRQoL. Which factors inuence HRQoL
most has not been thoroughly explored,
however.
Objective: Aim of this study is to explore
predictors of HRQOL results of patients
with asthma.
Materials and methods: 259 consecutive
patients (187 female and 72 male), aged
51,6 ( 18) (mean SD) years, diagnosed
with bronchial asthma and treated at the
Department of Allergology, Wroclaw Med-
ical University, Poland in 2008 and 2009
entered the study. Patients HRQoL was
assessed using two self-reported question-
naires: disease specic Asthma Quality of
Life Questionnaire (AQLQ), and Asthma
Control Test (ACT). A regression analysis
model was used to evaluate data from both
questionnaires.
Results: The results of regression analysis
in the AQLQ-symptoms-domain revealed
that major predictors for lower HRQoL
results were: concomitant diabetes, higher
score in ACT, active smoking and higher
number of hospitalizations (R2 = 0.507).
Variables that inuenced AQLQ-activity-
limitations-domain included: concomitant
diabetes, age >65, higher score in ACT,
higher number of hospitalizations, and
active smoking (R2 = 0.516). Scoring in
AQLQ-emotional-function-domain was
mainly inuenced by: concomitant diabe-
tes, low availability of allergologic outpa-
579

tient care, educational attainment, and
active smoking (R2 = 0.435). Finally, best
predictors of the results in AQLQ-environ-
mental-stimuli-domain were: concomitant
diabetes, length of duration of the disease
and low availability of allergologic outpa-
tient care.
Conclusions: HRQoL measured using AQL
Questionnaire is substantially reduced by
concomitant diabetes and asthma clinical
severity. Other important predictors of the
HRQoL in all four domains are: low avail-
ability of allergologic outpatient care, fre-
quent hospitalizations, and active smoking.
1535
Does drug compliance change in asth-
matic patients during pregnancy?
Yilmaz, I1; Oner Erkekol, F2; Celen, S3; Zayifoglu Karaca,
M3; Aydin, O1; Celik, G1; Misirligil, Z1; Mungan, D1
1
Department of Chest Diseases, Division of
Immunology and Allergy, Ankara University, School of
Medicine, Ankara, Turkey; 2Ataturk Chest Diseases &
Thoracic Surgery Training and Research Hospital, Unit
of Allergy, Ankara, Turkey; 3Dr. Zekai Tahir Burak
Womens Health Education and Research Hospital,
Department of Obstetrics and Gynecology, Ankara,
Turkey
Background: Pregnant women with asthma
are recommended to maintain optimal
therapeutic management during pregnancy.
However doubts about safety of asthmatic
drugs can affect asthmatic pregnant drug
compliance. The primary aim of this study
was to assess behavioral differences of drug
compliance among pregnant. Furthermore,
effects of asthmatic drugs used during
pregnancy on fetus and pregnancy/labour
complications were evaluated in this study.
Method: The study consisted of 153 preg-
nant women, 32 of them had asthma and
121 were healthy pregnant. Structured face
to face interviews were conducted after
delivery. The interviews included disease
characteristics, drug compliance and regu-
lar clinic visits before and during preg-
nancy. Furthermore, medical and
pregnancy history, pregnancy complica-
tions and outcomes, newborn characteris-
tics were recorded.
Result: The rate of regular asthma medica-
tion use was increased during pregnancy
(48%) when compared to pre-pregnancy
period (36%). Ninety percent of women
who used asthmatic drugs regularly before
pregnancy continued their medications dur-
ing pregnancy and 25% of irregular asth-
matic drug users before pregnancy began
to use asthmatic drugs systematically.
According to the patients evaluation
asthma status improved in 28%,
unchanged in 28% and worsened in 44%
of the subjects during pregnancy. Thirty-
ve percent of smokers quitted smoking
during pregnancy. Pregnant smokers had a
580
higher rate of low birth weight newborn
compared to nonsmoker pregnants. There
was no difference in terms of pregnancy/
labour complications, Apgar score, off-
spring malformation between asthmatic
and nonasthmatic pregnants.
Conclusion: Although regular use of
asthma drugs was increased during preg-
nancy this is still beyond what it should
be. The education of asthmatic women
particularly the ones who are in their fer-
tile ages should (deserve) a particular
attention to safe asthma medications in
pregnancy and asthma control as well as
maternal/perinatal outcomes even before
pregnancy. Smoking cessation strategies
should be followed as this is signicantly
associated with undesired fetal outcomes.
1536
A survey on asthma management: the
knowledge status about the usage of
inhaled medication devices by pediatri-
cians
Uysal, P1; Tuncel, T1; Arikan Ayyildiz, Z1; Firinci, F1;
Karaman, O1; Ellidokuz, H2; Uzuner, N1
1
Dokuz Eylul University Medical Faculty, Department of
Pediatrics, Division of Allergy, Izmir, Turkey; 2Dokuz
Eylul University Medical Faculty, Institution of
Oncology, Department of Preventive Oncology, Izmir,
Turkey
Background: Asthma control is currently
under expected levels and some of the
responsibility can be attributed to the med-
ical practitioners. We aimed to assess the
theoretical knowledge of the pediatricians
by performing a survey in a prospective
cross-sectional design during August and
September 2010, in Izmir.
Method: The pediatricians participating in
the study were surveyed with a question-
naire including 24 clinically relevant ques-
tions concerning doctors knowledge status
about asthma medications, usage of inhal-
ing devices, suitable treatment modalities
for different age groups, follow-up process
for asthma with regard to the recommen-
dations of national guidelines.
Result: We questioned about the controller
and reliever drugs: and only ve (6.1%)
and seventeen (20.5%) of the participants
gave the right answers, respectively. The
combination therapy of inhaled corticoster-
oids (ICS) and long-acting beta-agonists
(LABA) was found to be the most efcient
and most prescribed as the rst choice
controller drug for long-term asthma man-
agement in childhood. The preferred rst
choice of therapies for children under
5 years old who had frequent asthma exac-
erbations were low dose ICS or combina-
tion therapy of ICS with short acting
beta-agonists only during exacerbations in
one third of the participants, separately.
Allergy 66, Suppl. 94 (2011): 482642  2011 John Wiley & Sons A/S
Poster Session 65 The many faces of asthma
Most of the participants preferred conven-
tional nebulizator for ICS administration
and they used them in their daily clinics.
Thirty-four (42.5%) and 26 (32.1%) of the
participants believed that nebulizers and
metered dose inhalers (MDI) were the
most effective devices for drug administra-
tion respectively and only 21 (25.9%) of
them knew that their efciency was equal.
The most preferred drug administration
device was found as nebulizator for the
patients under 1 year old with a rate of
88.5%; MDI with valved holding chambers
(VHC) with face mask for children under
4 years of age was 76.9%. Only half of the
participants reported that they practiced
and instructed prescribed medicine on
demonstration device and observed the
inhalation technique of their patients.
Conclusion: Asthma management practice
of pediatricians falls short of the standards
recommended by international guidelines.
The participants own ideas about poor
knowledge and practice were the lack of
formal training during medical school and
residency and lack of postgraduate hands-
on-courses for training experience of the
skills.
1537
The allergy blog, twitter, facebook: An
educative experience using digital media
and social networks
Emerson, F; Rios, J; Pereira, N; Rios, J
Policlinica Geral do Rio de Janeiro, Clinica de Alergia,
Rio de Janeiro, Brazil
Background: Despite the increase of allergic
diseases over the last decades, the popula-
tion ignores the basic concepts, interfering
in their prevention and treatment. The
Allergy Blog (Blog da Alergia) has been
using digital media resources to offer new
forms of dialogue and patients ethical
enlightenment since 1996. This research
objectives to report the interactive educa-
tional experience, through the internet on
http://www.blogdalergia.com; and to nd
out, from the audiences questions, which
are the main areas of interest and where are
the greatest gaps in lay knowledge about
immune-allergic diseases.
Method: Research on Site Meters home
page to know the number of hits on the
site, Twitter and Facebook. Research on
Site Meter and Google Analytics to know
what were the main issues raised by the lay
public.
Result: Created by the Allergy Clinic of
the General Polyclinic of Rio de Janeiro,
Brazil, in 1996, Allergy Blog publishes edu-
cational texts and interacts with the visitors
through: (1) Comments posted on Blog, (2)
Doubts sent via e-mail, (3) Chat intended
Poster Session 65 The many faces of asthma
for short answers. Moreover, the blog has
been interconnected to Facebooks social
networks since 2009, where 100 links have
been published, totaling 380 followers and
publishing 1 200 tweets in Twitter, with
340 followers. By December 2010, the Blog
had totalled 1 462 075 viewed pages, keep-
ing an 1,817 daily average, 12 719 weekly
and 45 539 annual average visits. The blog
answered 4 256 e-mails and 2 200 com-
ments. However, 60 comments were
rejected because they referred to self-medi-
cation, sale or products promotion. The
most popular topics searched were: Corti-
costeroids (52.10%), pruritus (15.83%),
urticaria (7.36%), atopic dermatitis
(7.26%), allergic sinusitis (6.54%) and
cough (5.14%) among others (5.77%).
Most visitors are Brazilian, followed by
Portugal, USA, Japan, Germany and the
UK.
Conclusion: The use of digital media and
social networks may be a prime tool for
the education of allergic individuals, com-
munity dialogue and dissemination of cor-
rect information about the various aspects
of immune-allergic diseases. The discovery
of the lays greatest gaps and areas of
interest can be a guideline to improve new
educative actions.
1538
Pharmacists and asthma: survey among
120 pharmacists in urban and rural area,
comparison between 1999 and 2009
de Blay, F1; Meunier-Spitz, M1; Lefevre, H2; Rebotier, P1;
Bihler, A1; Heitz, C3
1
University Hospital of Strasbourg, Allergy Division,
Strasbourg, France; 2Department of Medical
Information, University Hospital of Strasbourg,
Strasbourg, France; 3Faculty of Pharmacy, University of
Strasbourg, Strasbourg, France
Pharmacist involvement in the treatment of
asthma has changed in the last ten years.
Though, few studies tried to measure those
changes.
Aim: To study pharmacists knowledge of
asthma treatment in urban and rural areas
between 1999 and 2009.
Material and methods: 120 pharmacists (60
licensed and 60 assistants) from the Bas-
Rhin (North part of Alsace) were ran-
domly selected on two occasions. The
2009s pharmacists were all different from
the 1999s ones. 86 of them accepted to
answer the same standardized question-
naire in 1999 and in 2009. Pharmacists
populations in 1999 and in 2009 were iden-
tical (age, gender, pharmacy location,...)
Emergency interventions from pharmacists
Allergy 66, Suppl. 94 (2011): 482642  2011 John Wiley & Sons A/S
for an asthma attack have been performed
by 10.6% of them in 1999 and 8.6% in
2009. 33% of them advised patients to go
back home in 1999 and 57% in 2009. They
gave advice to obtain a better compliance
(96% the two years) and explained how to
use an inhaler (96% in 1999 and 94% in
2009) in the same proportion. Ten years
later, 98% agreed to show the use of dif-
ferent inhalers instead of 63.9% in 1999
(P = 10-4) (MDI, Autohaler, Turbuhaler)
14 and 9% knew how to use an MDI
properly, 5 and 4% an Autohaler and 31
and 24% a Turbuhaler in 1999 and 2009,
respectively. Time spend per prescription
was the same in the two years (<15 min).
Conclusion: During the last 10 years, phar-
macist knowledge about asthma has not
increased signicantly. On the other hand
they are more likely to use an inhaler for a
demonstration. Training pharmacists about
asthma seems necessary.
1539
Does the use of longer or shorter educa-
tion material affect the success of one-to-
one asthma education by an allergy
nurse?
Epozturk, K; Koc, N; Abadoglu, O
Cumhuriyet University, Faculty of Medicine,
Department of Immunology and Allergic Diseases,
Sivas, Turkey
Background: For the optimal management
of asthma, the patient should acquire basic
knowledge and skills. The easiness to read
is important for a written education mate-
rial to reach success. We aimed to evaluate
the effects of longer and shorter education
materials on the information level of the
patient just after the session.
Method: An 11-item questionnaire was
applied face-to-face by an allergy nurse to
the patients admitted into the ward of
Allergic Diseases with a diagnosis of
asthma. Fifty three patients subsequently
received a 1-hour session of asthma educa-
tion with the use of a booklet prepared by
the national society of chest diseases. Forty
six patients were given a one-page informa-
tion text involving the answers of the ques-
tionnaire. At the end of the education, the
questionnaire was re-applied.
Result: The duration of the education ses-
sion with shorter material was averagely
26.2 1.7 min. The proportion of right
answers after the education session was sig-
nicantly higher in the group receiving the
shorter material (P < 0.05).
Conclusion: Asthma education can be done
with shorter and less time-consuming mate-
rials that include just key points of the dis-
ease features and treatment options.
1540
Role of serum IgG to aspergillus fumi-
gates by CAP in the diagnosis of allergic
bronchopulmonary aspergillosis in
patients with eosinophilic asthma
Park, M1; Lee, S1; Kim, K2; Nam, Y1; Um, S1; Lee, S1;
Son, C1
1
Dong-A University Hospital, Internal Medicine, Busan,
Republic of Korea; 2Dong-A University Hospital,
Laboratory Medicine, Busan, Republic of Korea
Background: Allergic bronchopulmonary
aspergillosis (ABPA) is associated with
hypersensitivity to Aspergillus fumigatus
(Af), occurring most commonly in atopic
patients with asthma and sometimes result-
ing in severe lung damage. However, there
have been some difculties in the diagnosis
of ABPA, because there were no reference
values based on individual serological
methods. We evaluated the role of the
serum IgG to Af using CAP system in the
diagnosis of ABPA in patients with eosino-
philic asthma.
Method: We used essential minimal criteria
for the diagnosis of ABPA. A total of 29
patients with more than 500 cell/Ul of
peripheral blood eosinophil were enrolled.
Prospective evaluation using skin prick test
(SPT) for Af, serum total and specic IgE
antibody to Af by CAP system, IgG anti-
body to Af by immunodiffusion or CAP
system and high resolution chest computed
tomography (HRCT) of chest were per-
formed.
Result: Among all 29 patients, just a single
patients (3.4%) compatible with ABPA
who had six items of the essential minimal
diagnostic criteria for diagnosis of ABPA.
IgG antibody to Af was not detected by
immunodiffusion in all of 26 patients who
had performed IgG assay by immunodiff-
fusion. By CAP system, IgG antibody lev-
els were shown from 0 to 40.6 (mgA/l).
These values were well correlated with IgE
to Af by CAP system (r = 0.40,
P = 0.04).
Conclusion: Specic serum IgG to Af by
CAP system is more sensitive, quantitative
and helpful in the diagnosis of ABPA in
patients with asthma. However, further
studies are required to dene reference val-
ues that can differentiate ABPA from
patients with asthma who sensitized with
Af.
581

Towards an integrated approach to asthma therapy
1541
Four weeks of ICS result in optimal
asthma control but not in full resolution
of airway inflammation in mild
asthmatics
Seys, S1; Dupont, L2; Decraene, A2; De Wandeleer, Y3;
Dilissen, E1; Ceuppens, J1; Bullens, D4
1
Katholieke Universiteit Leuven, Lab of Clinical
Immunology, Leuven, Belgium; 2Katholieke Universiteit
Leuven, Pneumology, Leuven, Belgium; 3University
Hospital Leuven, Respiratory Medicine, Leuven,
Belgium; 4Katholieke Universiteit Leuven, Woman and
Child, Leuven, Belgium
Background: The main goal in the treat-
ment of asthma is to obtain optimal clini-
cal asthma control and to prevent
exacerbations. It is however not clear
whether achieving asthma control and res-
olution of underlying airway inammation
correlate with each other.
Aim: To prospectively investigate the
changes in asthma control and inamma-
tion parameters during 4 weeks treatment
with 200 mg of budesonide compared to
placebo in mild asthmatics.
Method: 24 patients with mild persistent
asthma were recruited. Asthma control was
assessed by Asthma Control Test (ACT)
questionnaire. FeNO values were measured
by use of a chemiluminiscence analyser at
a ow rate of 50 ml/sec. Induced sputum
was obtained with an Ultranebb nebulizer
and sputum samples were processed as
described (1).
Result: ACT signicantly improved in the
ICS treated group (P = 0.034) but not in
the placebo group (P = 0.81). FeNO levels
tended to decrease but are still higher than
25 ppb in the majority (62.5%) of the ICS
treated patients (P = 0.11). FeNO levels
were signicantly lower after ICS compared
with values after placebo (P = 0.034). Per-
centages of sputum eosinophils slightly
increased after 4 weeks of placebo treat-
ment (P = 0.21), but tended to decrease in
the actively treated group (P = 0.094).
Sputum IL-5 mRNA levels remained
unchanged in both placebo (P = 0.5) and
the actively treated group (P = 0.58).
Table 1. For abstract 1541.
Placebo group ICS group Placebo group
baseline baseline after treatment
ACT 21 20.5 22
582
Poster Session 66
Conclusion: Despite clear clinical improve-
ment after 4 weeks of low dose ICS, there
were no remarkable changes in noninva-
sive markers of inammation such as
FeNO levels or sputum eosinophil percent-
ages. The majority of patients still had
persistent markers of airway inammation.
Also sputum cytokine levels were unaf-
fected by this treatment regimen. A med-
ium or high dose of ICS or a longer
period of treatment may be necessary to
obtain objective resolution of airway
inammation as assessed by FeNO or spu-
tum cell counts. The question remains
whether resolution of inammation is nec-
essary to maintain long term optimal
asthma control.
Reference: 1. Truyen E, Coteur L, Dilissen
E, Overbergh L, Dupont LJ, Ceuppens JL,
e.a. Evaluation of airway inammation by
quantitative Th1/Th2 cytokine mRNA
measurement in sputum of asthma
patients. Thorax. 2006 Mrt;61(3):202-8.
1542
Treatment with inhaled corticosteroids,
smoking and asthma control in real clini-
cal practice
Sergeeva, G; Emelyanov, A
Lung and Allergy Diseases Department, Saint-
Petersburg Medical Academy for Postgraduate Studies,
Saint-Petersburg, Russian Federation
Background: Treatment with inhaled corti-
costeroids (ICS) is a cornerstone of asthma
management. The aim of this study is to
assess the relationship between smoking,
ICS doses and asthma control in asthmatic
patients in real clinical practice in Saint-
Petersburg, Russia.
Methods: We analyzed three hundred case
record forms (CRF) of outpatients with
asthma (aged 1883 years, mean age
56 years, 25.7% males) treated with ICS
and followed up by pneumologists in 9 res-
idential areas in Saint-Petersburg, Russia.
Control of asthma was assessed according
to GINA (2006).
ICS group after Baseline P-values effect
treatment P-values of ICS
24 0.8 0.03
Allergy 66, Suppl. 94 (2011): 482642  2011 John Wiley & Sons A/S
Results: Among patients with asthma 36%
males were current smokers and 34% were
former smokers. In asthmatic females 15%
smoked currently and 12% were former
smokers. The majority of patients with
asthma were partly controlled. Females
had controlled asthma more frequent than
males (15% versus 8%, P < 0.05). Mean
doses of ICS in current (782.31 mcg) and
former (900.96 mcg) smokers were higher
than in non-smokers (733.68 mcg,
P < 0.05). Controlled asthma was less
common in current smokers (3%) com-
pared with former smokers (19%,
P < 0.05) and non-smokers (15%,
P < 0.05).
Conclusion: Smoking is common in asth-
matic patients and contributes to increase
of ICS doses to achieve controlled asthma.
1543
Ciclesonide in preventing asthma exacer-
bations
Kralimarkova, T; Staevska, M; Lazarova, C; Dimitrov, V;
Popov, T
Clinic of Allergy and Asthma, Medical University Sofia,
Sofia, Bulgaria
Background: Inhaled corticosteroids are
prescribed to prevent asthma exacerba-
tions, which pose imminent risks to the
patients and drive the deleterious process
of remodeling of their airways. The aim of
the proposed study was to document the
effectiveness and safety of ciclesonide,
recently registered for asthma maintenance
treatment in Bulgaria, in preventing
asthma exacerbations.
Method: The study was planned as an
observational open label real life study. It
involved 471 patients on ciclesonide (mean
age 42 18/SD/years, 285 women) and
385 on all other ICS prescribed in Bulgaria
(mean age 44 19 years, 220 women).
The recruitment of patients was made by
specialists from 36 different practices
across Bulgaria. Patients were assessed on
2 visits one month apart. All outcomes of
the study were derived from an extended
Asthma Control Questionnaire, adapted to
the specic Bulgarian environment. The
scores from the questionnaires before and
after treatment were analyzed to assess the
effectiveness of Ciclesonide treatment and
further compared with the scores of the
Poster Session 66 Towards an integrated approach to asthma therapy
active controls on alternative treatments.
Physician also reported whether asthma
exacerbations occurred during the observa-
tional period.
Result: Out of the patients on ciclesonide,
396 subjects stated that their condition
improved, 68 thought that it did not
change, and 7 considered their condition
deteriorated. Furthermore, 397 rated ci-
clesonide better, 60 similar, and 12 did not
think it differed from the previous ICS
treatment they were taking. A whole range
of adverse events (AE) was reported by the
patients without prompting for ciclesonide
and all the other ICS. When comparing
the specic adverse events, related to the
irritation in the throat, there was a highly
signicant difference in favour of cicleso-
nide. Physicians noted that 29 (6.2%) of
subjects on ciclesonide had mild exacerba-
tions during the treatment course, com-
pared with 109 (28.3%) of patients on
other ICS, P < 0.001.
Conclusion: Ciclesonide was proven effec-
tive as asthma maintenance treatment in
an open label real life study. The prefer-
ence given to it by patients may be
prompted by the promise of the new
drug effect, which may also have
improved patients compliance. The latter
fact may also explain the much lower exac-
erbation rate with ciclesonide.
1544
The efficacy of single-high dose inhaled
corticosteroid treatment on leukotrienes
and 8-isoprostane levels evaluated with
exhaled breath condensate in children
with asthma exacerbation: preliminary
results
Keskin, O1; Keskin, M2; Gogebakan, B3; Uluca, U3;
Kucukosmanoglu, E1; Ozkars, M1; Bayram, H4; Coskun,
Y2
1
Gaziantep University Hospital, Pediatric Allergy,
Gaziantep, Turkey; 2Gaziantep University Hospital,
Pediatrics, Gaziantep, Turkey; 3Gaziantep University
Hospital, Gaziantep, Turkey; 4Gaziantep University
Hospital, Pulmonology, Gaziantep, Turkey
Background: Exhaled breath condensate
(EBC) is a completely noninvasive method
for the collection of airway secretions.
There is no study investigating the efcacy
of single high dose inhaled corticosteroid
treatment and also a six days of high dose
inhaled corticosteroid treatment on the
measure of exhaled Cys-LTs and 8-isopros-
tane levels in children with asthma exacer-
vation. We aimed to assess the efcacy of
single high dose inhaled corticosteroids on
the measure of exhaled Cys-LTs and 8-iso-
prostane levels in children with asthma ex-
acervation.
Method: Fortyve children with moderate-
severe asthma exacervation were evaluated
with asthma score, pulmonary function
Allergy 66, Suppl. 94 (2011): 482642  2011 John Wiley & Sons A/S
tests and exhaled Cys-LTs and 8-isopros-
tane levels. Exhaled breath condensate was
collected before treatment, after 4 h of
treatment with inhaled 4000 lg uticasone
propionate (FP) and after six days of treat-
ment with inhaled FP 1000 lg/daily. Cys-
LTs and 8-isoprostane concentrations were
determined using a specic immunoassay
kit.
Result: Single high dose inhaled FP treat-
ment induced a signicant improvement in
asthma score (P < 0.05) and pulmonary
function tests (P < 0.05). After 4 h of
treatment with inhaled FP (4000 lg) there
was a signicant decrease in exhaled Cys-
LTs concentrations (P = 0.019). Cys-LTs
concentration in EBC was decreased much
more signicantly after six days of treat-
ment with inhaled FP compared to exhaled
Cys-LTs levels before treatment
(P = 0.001). There was a marginally sig-
nicant decrease in 8-isoprostane concen-
trations in the EBC after six days of
treatment with inhaled FP (1000 lg/day)
compared to exhaled 8-isoprostane levels
measured after 4h of treatment
(P = 0.075). The percentage reduction in
Cys-LTs and in 8-isoprostane after inhaled
FP correlated signicantly with baseline
Cys-LTs levels (P = 0.037, r = 0.45) and
with baseline 8-isoprostane levels
(P = 0.038, r = 0.47) respectively. There
was a signicant correlation between EBC
Cys-LTs and 8-isoprostane levels in chil-
dren with asthma exacervation (P = 0.001,
r = 0.57).
Table 1. For abstract 1544.
Baseline Cys-LTs (pg/ml) 31 (17, 68)
Cys-LTs (pg/ml) (After 12 (4, 32)
4 h of single high dose
inhaled corticosteroid
treatment)
Cys-LTs (pg/ml) (After 4 (4, 15)
6 days of high dose
inhaled corticosteroid
treatment)
Baseline 8-isoprostane 25 (10, 59)
(pg/ml)
8-isoprostane (pg/ml) 33 (13, 67)
(After 4 h of single
high dose inhaled
corticosteroid treatment)
8-isoprostane (pg/ml) 12 (8, 39)
(After 6 days of high
dose inhaled
corticosteroid
treatment)
Conclusion: We have shown antiinamma-
tory efcacy of single high dose inhaled
corticosteroids on exhaled Cys-LTs in chil-
dren with asthma exacervation. Exhaled 8-
isoprostane level, a marker of oxidative
stress, was not decreased with single high
dose inhaled corticosteroid treatment but
there was a marginally signicant decrease
in exhaled 8-isoprostane levels after 6 days
of high dose inhaled corticosteroid treat-
ment.
1545
Predictive risk factors for relapse after
cessation of inhaled corticosteroids in
well-controlled childhood asthma
Akturk, H; Karakoc-Aydiner, E; Keles, S; Yildiz, A; Akkoc,
T; Ozdemir, C; Barlan, I; Bahceciler, N
Marmara Medical Faculty, Division of Pediatric Allergy
and Immunology, Istanbul, Turkey
Background: There is no consensus on the
duration of inhaled corticosteroid (ICS)
treatment in controlled asthma. Hereby,
we evaluated inammatory, clinical and
functional parameters at the time of ICS
withdrawal to predict asthmatic children
who will experience recurrence of symp-
toms.
Method: Forty children asymptomatic for
at least 3 months on low dose ICSs were
enrolled. Demographic and clinical features
were recorded. After ICS withdrawal,
patients were evaluated every 2 month for
6 months. At enrolment symptom/medica-
tion diary, pulmonary function parameters,
methacholine provocation testing, peripher-
ic blood eosinophilia (PBE), serum total
and allergen-spesic IgE levels and skin
prick testing were performed. In a sub-
group of patients house dust mite allergen
(Der p 1) and phytohemaglutinin induced
secretion of IL-5, IL-13, IFN-c, IL-10
from blood mononuclear cells were mea-
P
sured at baseline. Follow up visits included
assessment of symptom/medication diary,
0.019 pulmonary function test (PFT) and physi-
cal examination. Patients with asthma
exacerbations were evaluated by physical
examination, PFT and PBE.
0.001
Result: Twenty-one out of 40 patients
(52.5%) experienced recurrence of asthma
symptoms. Baseline bronchial hyperrespon-
siveness (BHR) was found to be a risk fac-
tor for asthma exacerbation. FEF25-75
values tended to be lower throughout the
study in patients with symptom recurrence.
Moreover, rhinitis associated symptoms
were higher during the study. Finally, Derp
0.075
induced IL-5 and IL-13 levels were higher
and IFN-c levels were lower in house-dust-
mite sensitized patients with recurrence of
asthma.
Conclusion: We concluded that before the
cessation of ICSs in children with well-con-
trolled asthma BHR should be evaluated
and symptoms of rhinitis should be con-
trolled. Tendency of decrease in FEF25-75
values during follow-up may point out the
recurrence of asthma symptoms.
583

1546
Effectiveness of fluticasone propionate
and rare adverse effects in preschoolers
with asthma
Bajraktarevic, A1; Maglajlija, S1; Penava, S1; Begovic,
B2; Selimovic, A3; Dzinovic, A3; Guzin, Z4; Frankic, T5;
Gutic, J6; Djulepa Djurdjevic, A6; Roncevic, Z7;
Sporisevic, L8; Prnjavorac Rakic, B9
1
Pediatrics Department, Public Health Institution of
Canton Sarajevo, Sarajevo, Bosnia and Herzegovina;
2
Clinical Medical Center Sarajevo, Clinical
Pharmacology, Sarajevo, Bosnia and Herzegovina;
3
Department for allergology and pulmonology,
Pediatrics Clinic Sarajevo, Sarajevo, Bosnia and
Herzegovina; 4Emergency Department, City Hospital
Mostar, Mostar, Bosnia and Herzegovina; 5Pharmacy
Faculty Sarajevo, Department for Clinical
Pharmacology, Sarajevo, Mostar, Bosnia and
Herzegovina; 6Emergency Department, General Hospital
Sarajevo, Sarajevo, Bosnia and Herzegovina;
7
Cardiology Department, Pediatrics Hospital Mostar,
Mostar, Bosnia and Herzegovina; 8Pediatrics
Department, First Medical Aid of Canton Sarajevo,
Sarajevo, Bosnia and Herzegovina; 9Pediatrics
Department, Pediatrics Health Center Tesanj, Tesanj,
Bosnia and Herzegovina
Background: Inammation is recognized as
an important component in the pathogene-
sis of asthma. Fluticasone propionate is a
synthetic, triuorinated glucocorticoid with
potent anti-inammatory activity. This
medication must be used regularly to pre-
vent the wheezing and shortness of breath
caused by asthma or obstructive bronchitis,
or some rare types of emphysema in chil-
dren.
Aim: To determine if the early use of
inhaled uticasone propionate in wheezy
preschooler children older helps to prevent
loss of lung function and progression of
asthma later in school-childhood.
Method: This study was a randomized,
double-blind, placebo-controlled prospec-
tive trial using other therapy from golden
rules and guidelines GINA modied for
children. Spirometry and induced sputum
for differential cell counts and albumin,
2-macroglobulin and blood eosinophil
and, interleukins factor levels were
obtained before treatment and two, six
and twenty four hours after treatment in
children with asthma in age between 2 to
7 years of ages during rst decade of new
millennium.
Result: When glucocorticoids are discon-
tinued, asthma stability may persist for
several days or longer. The total clearance
of uticasone propionate is high, with
renal clearance accounting for less than
0.02% of the total. This medication does
not work immediately, because it is preven-
tive and prolonged action. The most chil-
dren (99.5%) older than two years in
Bosnia and Herzegovina using this medica-
tion do not have serious side effects. Pre-
schoolers with recurrent wheezing or
asthma had less wheezing or asthma exac-
erbations and improve their symptoms and
lung function during treatment with
584
Poster Session 66 Towards an integrated approach to asthma therapy
inhaled corticosteroids in the most cases
(98%) in our ten years study.
Discussion: The precise mechanisms of glu-
cocorticoid action in asthma are unknown.
The safety and effectiveness of Fluticasone
propionate inhalation aerosol in children
below two years of age have not been
established.
Conclusion: Fluticasone propionate inhala-
tion aerosol is contraindicated in the pri-
mary treatment of status asthmaticus or
other acute episodes of asthma where
intensive measures are required. The
authors concluded that inhaled uticasone
given twice daily over a 6-month period
improved asthmatic symptoms and had no
signicant adverse effects on growth.
1547
Inhaled steroids and caries, dental plaque
and salivary flow in asthmatic adoles-
cents
Santos, N1; Jamelli, S2; Medeiros, D3; Baracho Filho,
C4; Rizzo, J5; Sarinho, E6
1
Federal University of Feira de Santana, Brazil,
Odontology Department, Feira de Santana, Brazil;
2
Federal University of Pernambuco, Recife, Brazil,
Professor of Post-graduate courses in Child and
Adolescent Health, Recife, Brazil; 3Federal University of
Pernambuco, Brazil, Allergy and Immunology, Recife,
Brazil; 4Federal University of Pernambuco, Pediatrics,
Recife, Brazil; 5Federal University of Pernambuco,
Recife, Brazil, Professor of Post-graduate course in
Health Sciences, Recife, Brazil; 6Federal University of
Pernambuco, Recife, Brazil, Professor of Post-graduate
courses in Child and Adolescent Health and Health
Sciences at the Federal University of Pernambuco,
Brazil
Background: A number of studies have
reported that inhaled steroids may cause a
greater incidence of caries, reduced salivary
ow, changes in saliva composition and an
increased frequency of dental plaque, prob-
ably through alterations in the oral micro-
biota. The objective was to compare the
frequency of caries, dental plaque and non-
stimulated salivary ow rate amongst asth-
matic adolescents using inhaled steroids
and non-asthmatic adolescents, as well as
the salivary biochemical parameters (pH
and leukocytes) in both groups.
Method: This research has a descriptive
cross-sectional design to compare dental
health of 40 asthmatics on inhaled steroids
and 40 non-asthmatics adolescents (median
age 13 years).
Result: It was found a higher number of
tooth surfaces affected by dental caries
(median 4 versus 1.5), and more dental pla-
ques (median 70.5 versus 60.7) among asth-
matic adolescents. They also had a
signicantly higher frequency of salivary
leukocytes. The non-stimulated salivary
ow was similar in both groups.
Conclusion: The results suggest an associa-
tion between the use of inhaled steroid and
an increased risk of dental caries and bac-
Allergy 66, Suppl. 94 (2011): 482642  2011 John Wiley & Sons A/S
terial plaque which calls for special atten-
tion of these patients by doctors and
dental health professionals.
1548
Decreased interleukin-10 levels in
induced sputum of asthmatic patients
despite chronic glucocorticoid treatment
Moniuszko, M1; Jeznach, M1; Dabrowska, M2; Bodzenta-
Lukaszyk, A1
1
Department of Allergology and Internal Medicine,
Medical University of Bialystok, Bialystok, Poland;
2
Department of Hematological Diagnostics, Medical
University of Bialystok, Bialystok, Poland
Background: Despite some conicting data,
interleukin )10 (IL-10) levels have been in
general reported to be reduced in the air-
ways of asthmatic patients. On the other
hand, glucocorticoids (GC) bear the poten-
tial to enhance the secretion of endogenous
IL-10. Recently, however, we demonstrated
that GC are capable of decreasing the
expression of IL-10 receptor. In the current
study we investigated the inuence of
inhaled and oral GC treatment on IL-10
levels in induced sputum of twenty-two sta-
ble asthmatic patients.
Methods: Induced sputum IL-10 levels
were determined by ELISA and subse-
quently correlated with clinical status, cur-
rent treatment regimen and measurements
of the other markers of airway inamma-
tion in asthma.
Results: We found that GC-treated asth-
matic patients presented with similar to
GC-naive asthmatics median airway IL-10
levels (6.84 pg/ml [interquartile range,
IQR, 1.0913.1 versus 8.04 pg/ml [IQR,
4.2022.9], P > 0.05). Following more
detailed analysis, this pattern held true as
for patients who were chronically treated
with both inhaled and oral GC (6.84 pg/ml
[IQR, 0.8413.4] versus 8.04 pg/ml [IQR,
4.2022.9], P > 0.05), as for those who
were receiving only inhaled GC (6.88,
[IQR, 1.5613.4] versus 8.04 pg/ml [IQR,
4.2022.9], P > 0.05). There was no statis-
tically signicant difference in induced
sputum IL-10 levels between those two
GC-treated groups of patients (P > 0.05).
Conclusions: Our data suggest that, in con-
trary to expectations, GC treatment
doesnt have to result in increasing IL-10
levels in asthmatic airways. Thus, the inter-
play between GC and IL-10 in asthma
might be more complex than initially
thought.
Poster Session 66 Towards an integrated approach to asthma therapy
1549
Comparing the safety and efficacy of flu-
ticasone/formoterol combination therapy
in a single inhaler at low and medium
doses, with fluticasone and formoterol
concurrently or alone, in asthma patients
Hochhaus, G1; Kaiser, K2
1
University of Florida, Gainesville, United States;
2
SkyePharma AG, Muttenz, Switzerland
Background: A Phase 2, randomised, pla-
cebo-controlled, 6-way crossover, single-
dose exposure study to evaluate the safety
and efcacy of uticasone/formoterol com-
bination therapy (FLUT/FORM 100/10 lg
and 250/10 lg b.i.d.) in a single metered
dose inhaler when compared with utica-
sone propionate (FLUT 250 lg b.i.d.) and
formoterol fumarate (FORM 12 lg b.i.d.)
concurrently or alone in patients with mild
to moderate asthma.
Method: Patients included men and women
aged 18 to 65 years with a history of
asthma 12 months, FEV1 of 4080%
(inclusive) of predicted normal and FEV1
reversibility (15% within 30 min after
salbutamol), who maintained asthma medi-
cation at a stable dose for 2 weeks before
and during screening. Patients in each
group received 4 of the 6 treatments
administered on separate study days with a
washout of 3 to 14 days.
Result: 64 patients were randomised.
Comparable efcacy was observed for
treatment with FLUT/FORM 100/10lg or
250/10 lg compared with FLUT
250 lg + FORM 12 lg. Superior efcacy
was observed with both doses of FLUT/
FORM compared with FLUT or placebo
(for change in actual FEV1 (P < 0.001),
% change in FEV1 (P < 0.001), area
under FEV1 curve (P < 0.001), FEV1
max (P < 0.001) and FEV1 % predicted
normal values (P < 0.001) at 12 h).
Change from baseline for PEFR, FVC
and FEF25-75% improved with all active
treatments. The onset of clinical effect was
similar with all active treatments (range
4.3 to 6.9 min) and was statistically signi-
cantly faster with FLUT/FORM 100/
10 lg and 250/10 lg (P = 0.002 for both)
than placebo. For responders, the clinical
effect (FEV1 >15% above baseline) for
FLUT/FORM was maintained for 13
15 h post-dose. No serious AEs were
reported. The most frequently reported
AEs were headache, asthma and dizziness.
The frequency of asthma exacerbations
reported as an AE was similar between
the active treatment groups; 4 patients
(10%) in the FLUT/FORM 100/10 lg
group; 4 (11%) in the FLUT/FORM 250/
10 lg group; 5 (12%) in the FLUT
250 lg + FORM 12 lg group; 3 (8%) in
the FLUT 250 lg group; and 5 (13%) in
the FORM 12 lg group. In the placebo
Allergy 66, Suppl. 94 (2011): 482642  2011 John Wiley & Sons A/S
group, 3 patients (8%) reported asthma
exacerbations as AEs.
Conclusion: The efcacy and safety proles
of FLUT/FORM at low and medium
doses were similar to those of FLUT and
FORM administered concurrently in
patients with mild-to-moderate asthma.
These results provide further support for
FLUT/FORM as a potential treatment for
asthma.
1550
Pharmacokinetics of fluticasone/formo-
terol combination therapy in a single
inhaler compared with fluticasone and
formoterol concurrently or alone in
healthy subjects: results of two open
label studies
Hochhaus, G1; Kaiser, K2
1
University of Florida, United States; 2SkyePharma AG,
Switzerland
Background: Presented here are the results
of two randomised, open-label, parallel
group, multiple dose exposure studies com-
paring the pharmacokinetics of uticasone/
formoterol combination therapy (FLUT/
FORM) in a single inhaler with uticasone
(FLUT) and formoterol (FORM) adminis-
tered concurently or alone (Study 1) or
FLUT adminstered alone (Study 2).
Method: Subjects were healthy male or
female volunteers aged 1945 years who
demonstrated a forced expiratory volume
in 1 s (FEV1) > 80% of predicted normal
value. The studies compared FLUT/FORM
(100/10 lg BID and 250/10 lg BID, Study
1); 250/10 lg BID, Study (2) with FLUT
(250 lg BID) and FORM (10 lg BID,
Study 1) or just FLUT (250 lg BID, Study
2). Each of the subjects received multiple
doses of one of the 5 treatments (Study 1)
or 2 treatments (Study 2).
Result: For Study 1, 50 subjects were
enrolled, 47 completed; for Study 2, 36
subjects were enrolled, 35 completed. The
mean Cmax and AUCtau of FLUT follow-
ing a single dose was lower for FLUT/
FORM (250/10 lg) compared with FLUT
alone or plus FORM in Study 1. However
after BID dosing for 1 week, mean Cmax
and AUCtau were comparable between
FLUT/FORM (250/10 lg) and FLUT but
lower than FLUT plus FORM. For Study
2 the mean Cmax and AUCtau of FLUT
following both a single dose and BID dos-
ing for 7 days, were very similar for
FLUT/FORM and FLUT given alone, as
indicated by differences of <20% between
values while the observed variabilities were
larger. Treatments in both studies showed
some FLUT accumulation following BID
dosing for 1 week, consistent with litera-
ture values. For FORM (Study 1), on Day
1 when subjects were not at steady state,
the mean % of dose in urine (%Fe) was
comparable for FLUT/FORM (250/10 lg),
FLUT plus FORM, and FORM alone and
was less than half that of FLUT/FORM
(100/10 lg). Following BID dosing for
1 week, the mean %Fe for FORM was
comparable for FLUT/FORM (250/10 lg),
FLUT plus FORM, and FORM alone and
only slightly lower than that following
FLUT/FORM (100/10 lg). BID dosing for
7 days resulted in higher mean %Fe on
Day 7 for all treatments except FLUT/
FORM (100/10 lg) where mean %Fe
decreased compared with Day 1.
Conclusion: The pharmacokinetic prole of
FLUT/FORM at steady state was similar
to that of FLUT administered separately,
but FLUT/FORM had a lower systemic
exposure compared with the individual
components given concurrently
(FLUT + FORM).
1551
Fluticasone propionate and Budesonide
do not stop the in vitro effect of
pro-inflammatory cytokines on
pulmonary fibroblasts phenotype
Panaitescu Bunu, C; Matis, B; Gavriliuc, O; Tanasie, G
University of Medicine and Pharmacy Victor Babes
Timisoara, Physiology and Immunology, Timisoara,
Romania
Background: Asthma is characterized by
inammation and the activation of tissue
remodeling that result in structural altera-
tions such as airway wall thickening. Dif-
ferent drugs are commonly used or
potentially useful in the treatment of
asthma, corticosteroids (CS) and their ben-
ecial effects being often ascribed to their
anti-inammatory properties. Recently the
effects of corticosteroids kept attention to
their potential role on tissue remodeling.
This study was designed to evaluate the in
vitro effect of proinammatory cytokines
on the airway broblast phenotype and the
role of corticosteroids in this process.
Method: Pulmonary broblasts (PF) were
incubated in vitro with TGF-beta1, TGF-
beta2, IL-13, IL-4 and with a mixture of
all these factors. Differentiation to myo-
broblasts was characterized by the expres-
sion of alpha-smooth muscle actin (a-
SMA), myosine, desmine using immunohis-
tochemistry. We investigated the effects of
CS uticasone propionate (FP) and budes-
onide (B) on the production of a-SMA by
PF with or without stimulation by proin-
ammatory cytokines. PF were pretreated
with CS for 24 h, and then cultured with
or without proinammatory cytokines and
CS in increasing concentrations (10)12 M,
10)9 M and 10)6 M) for 1 week, followed
by harvesting and processing for immuno-
histochemistry.
585

Result: The assay of broblasts pulsed
with factors showed a positive reaction for
a-SMA expression. The percentage of posi-
tive cells was higher in broblasts pulsed
with all factors together (P = 0.008),
tended to be lower in broblasts pulsed
with TGF-beta1 or TGF-beta2 and IL-4
seemed to be the most pro-brogenic fac-
tor, comparing to negative control
(P = 0.0001). Corticosteroids pretreatment
didnt inuence signicantly the a-SMA
expression in pulsed broblasts.
Conclusion: These data suggest that CS
control airway remodeling through a
reduction of inammation, but such an
approach is partly successful and does not
conrm the hypothesis of acting directly
on pulmonary resident cells. Although cor-
ticosteroids do affect some aspects of air-
way remodeling, they are obviously far
from totally protective to long-term airway
damage, but precisely how effective they
are or can be needs to be better dened.
1552
Therapeutic efficacy of the metered dose
inhaler formulation of budesonide/formo-
terol in Hungarian children with asthma
Endre, L1; DuBuske, L2
1
Heim Pal Childrens Hospital, Budapest, Hungary;
2 study to receive FLUT/FORM 250/10 lg
Immunology Research Institute of New England,
Gardner, MA, United States
Background: Children with moderate to
severe asthma may require combination
therapy including an inhaled corticosteroid
and long acting beta-2 agonist for optimal
therapeutic responses. To date there have
been no reports investigating the impact of
combination therapy with formoterol and
budesonide in asthmatic children in Hun-
gary.
Method: In an outpatient clinic in Buda-
pest, Hungary in 2008, 183 asthmatic chil-
dren (91 boys, 53 girls, aged 418 years)
were treated with a metered dose formula-
tion of budesonide/formoterol (80/4.5 lg,
or 160/4.5 lg) for one year. Clinical ef-
cacy and adverse effects were assessed.
Result: During the year long observation
period only one child treated with the
metered dose formulation of budesonide/
formoterol needed hospitalization because
of dyspnea. 8 patients suffered exacerba-
tions lasting 2 to 3 days, but these children
responded well to symptomatic treatment
at home. No children reported increased
nocturnal awakenings because of dyspnea.
No patients developed oral Candida infec-
tions. The treated children showed no evi-
dence of growth retardation, in
comparison with healthy children.
Conclusion: Treatment with budesonide/
formoterol combination therapy in a
metered dose inhaler was effective and well
586
Poster Session 66 Towards an integrated approach to asthma therapy
tolerated by Hungarian asthmatic children.
This is the rst report documenting the
efcacy of this therapeutic approach in
Hungarian children.
1553
Effect of high dose inhaled fluticasone/
formoterol combination therapy com-
pared with its separate components on
the hypothalamo-pituitary-adrenal (HPA)
axis function
Priestley, A1; Woodward, J2; McIver, T2; Mundin, G2
1
LCG BioScience, Bourn, Cambridge, United Kingdom;
2
Mundipharma Research Limited, Cambridge, United
Kingdom
Background: A new asthma therapy com-
bining uticasone propionate and formo-
terol fumarate in a single HFA pMDI
(FLUT/FORM) has been shown to be at
least as effective as equivalent doses of the
individual components administered con-
currently (FLUT + FORM; pMDI). This
study compared the effect on HPA axis
function of treatment with FLUT/FORM
with FLUT+FORM using 24-hour uri-
nary free cortisol measurements.
Method: Healthy volunteers (n = 48) were
randomised in an open-label, 2-treatment,
parallel-group, multiple-dose exposure
(two actuations b.i.d.) or FLUT
250 lg + FORM 12 lg (each two actua-
tions b.i.d.) with a spacer device (Aero-
Chamber Plus, GSK). Subjects were
dosed on the morning of Day 1, then twice
daily from the morning of Day 3 to Day
29. Urine samples for urinary free cortisol
measurement were taken over two 24-hour
periods (Days )2 to )1 and Days 27 to
28). Basal serum cortisol and ACTH stim-
ulation were measured pre-dose on Days 1
and 29. Serial blood and urine samples for
pharmacokinetic analysis were collected on
Days 1 and 29.
Result: There was a more pronounced
decrease in mean 24-hour urinary free corti-
sol levels (corrected for creatinine) at the
end of treatment with FLUT+FORM
(4 nmol/24 h versus 14 nmol/24 h at base-
line) than FLUT/FORM (7 nmol/24 h
versus 13 nmol/24 h at baseline). Pharmaco-
dynamic bioequivalence could not be con-
cluded between the treatments at the end of
treatment since the 95% CIs for the ratio
did not fall within the range 0.52.0 for
transformed values (ratio: 1.9; 95% CIs 1.5,
2.6) or untransformed values (ratio: 1.8;
95% CIs: 1.4, 2.2). HPA axis suppression
was more pronounced in subjects taking the
individual components. Basal serum cortisol
results supported this nding, with values
on Day 1, Day 29 and post-study of 399,
272 and 412 nmol/l for FLUT/FORM and
430, 154 and 390 nmol/l for FLUT+
Allergy 66, Suppl. 94 (2011): 482642  2011 John Wiley & Sons A/S
FORM. ACTH stimulation test responses
were similar for both treatment groups on
Day 1 and Day 29, indicating no signicant
adrenal insufciency was induced during the
4 week treatment period. The bioavailability
of uticasone and formoterol in the FLUT/
FORM group was lower than in the
FLUT + FORM group on Day 1 (ratios of
37% and 17%, respectively) and at steady
state on Day 29 (ratios of 67% and 75%,
respectively).
Conclusion: FLUT/FORM elicited less
HPA axis suppression than FLUT + -
FORM, with a corresponding bioavailabil-
ity of uticasone that was 67% of that
from the individual component at steady
state.
1554
A systematic review on the efficacy and
safety of statin use in asthmatic patients
Silva, D; Couto, M; Moreira, A; Delgado, L
Allergology and Clinical Immunology Department, S.
Joao University Hospital, Porto, Portugal
Background: 3-hidroxy-3-methylglutaryl-
coenzime A reductase inhibitors are known
by their lipid-lowering effects and have an
established role in the treatment of athero-
sclerotic disease. They also have anti-
inammatory and immunomodulatory
properties that could affect allergic dis-
eases. The aim of this study is to review
the evidence about the efcacy and safety
of statins use on asthmatic patients.
Method: Systematic review of the literature
through a search in the National Guideline
Clearinghouse, Cochrane, Scopus and Pub-
med Medline of all the articles published
using the MESH terms: Hydroxymethyl-
glutaryl-CoA Reductase Inhibitors, Hyper-
sensitivity, Contact Dermatitis, Asthma,
Allergy and Immunology, Seasonal Aller-
gic Rhinitis and Immunomodulation.
Expert opinion, classical reviews, animal
studies and all articles including subjects
not exposed to an intervention with statins
were excluded. Centre for Evidence Based
Medicine Levels of Evidence was used to
classify levels of evidence and grade of rec-
ommendation.
Result: 523 articles where obtained and 41
were selected based on title and abstract. Of
these, 12 articles were included: 3 random-
ized control trials (RCT), 3 case control
studies, and 6 case reports. Pooling the data
from RCT trials, no statistical signicant
benets with the use of statins appeared in
asthmatic patients, in relation to patient
oriented (asthma control and quality of life
questionnaire, steroid-sparing effects) and
disease oriented outcomes (peak expiratory
ow, forced expiratory volume 1s, methaco-
line hyperresponsiveness, fraction of
Poster Session 66 Towards an integrated approach to asthma therapy
exhaled nitric oxide and sputum eosinoph-
ils). One retrospective case control study in
asthmatic patients showed a potential haz-
ardous association in asthma control. Case
reports of adverse reactions included: 2
drug induced hypersensitivity pneumonitis,
2 eczematous lesions, 1 chronic urticaria
and 1 nasal polyps associated to statin use.
Conclusion: Statins appear not to be effec-
tive in asthma control and no steroid-spar-
ing benet was found in asthma treatment.
There is limited evidence of hazardous con-
sequences of statin use in asthmatic
patients. Given the prevalence of statin use
and asthma disease more studies are
needed to conrm these data.
1555
Quality of treatment of asthma in
primary health care facilities in Saint-
Petersburg, Russia
Tsukanova, I1; Emelyanov, A1; Fedoseev, G2;
Sergeeva, G3; Lisitsyna, N2; Bakanina, L2; Nikitina, E2
1
Department of Respiratory Disease, Postgraduate
Medical Academy, Saint-Petersburg, Russian
Federation; 2Pavlov Medical University, Saint-
Petersburg, Russian Federation; 3Postgraduate Medical
Academy, Saint-Petersburg, Russian Federation
Background: This study was performed to
assess the quality of diagnosis and treat-
ment of asthma in primary health care
facilities.
Method: Case record forms (CRF) of 397
outpatients (30.5% male), 18 to 84 years
old (mean age 55 years), with asthma were
reviewed in 13 outpatient departments in 7
residential areas in Saint Petersburg. We
analyzed controller regimens of anti-
inammatory medications of patients with
persistent asthma.
Result: During the past 12 month inhaled
corticosteroids (ICS) were used in 36.2%,
ICS + LABA in 5.9%, xed combina-
tion (ICS+LABA) - in 17%, (ICS+LA-
BA)+ICS - in 3% of patients. Oral
steroids + inhaled corticosteroids were
prescribed to 13.5% and 2.7% of patients
in persistent asthma were treated only oral
steroids for maintenance therapy. In 2.7%
were used cromones. Most patients
(68.3%) used SABA, long acting oral theo-
phyllines were dispensed to14.6%. Antileu-
kotrienes were not prescribed. Allergen
specic immunotherapy were used only in
1.1% of patients.
Conclusion: Quality of anti-asthma con-
troller therapy in primary health care is
not sufcient and should be improved.
Allergy 66, Suppl. 94 (2011): 482642  2011 John Wiley & Sons A/S
1556
The significance of the use of acaricides
in fluctation of symptoms of the asth-
matic patients sensitive to house dust
mites
Pechlivanidis, T1; Sichletidis, L2; Damianakos, D1
1
Social Insurance Services (IKA), Thessaloniki,
Pulmonary Clinic, Thessaloniki, Greece; 2Aristotele
University of Thessaloniki, Greece, Pulmonary Clinic,
Thessaloniki, Greece
Background: The purpose of this study
was the evaluation of the signicance of
the correct use of acaricide products in the
improvement of the symptoms of the asth-
matic patients allergic to HDM.
Method: 30 asthmatic patients, who were
sensitive to HDM were equally divided in
two groups and they were studied. All
patients were receiving a constant, if possi-
ble, therapy (inhalations of a b2-stimu-
lant). The rst group additionally took
preventive measures against HDM by sys-
tematic use of an acaricide product (Acaro-
san), contrary to the second group which
took no preventive measure at all. The
methodology of all patients follow-up
included: a) Measuring of PEFR on a daily
basis (morning-evening) and b) Daily
recording of the probable symptoms
(cough, dyspnoea) as well as the variations
in the therapeutic scheme followed by the
patients. The duration of the follow-up
was one calendar year.
Result: The uctuation of the mean values
of the patients PEFR in both groups did
not show signicant statistical differences
from month to month (f = 0.12,
P = 0.99). The respiratory symptoms of
the rst group did not show signicant dif-
ferences from month to month (f = 1.16,
P = 0.31) contrary to those of the second
group, which showed signicant statistical
differences (f = 9.67, P = 0.00). The vari-
ation of the inhaled doses of the b2-stimu-
lant in the rst group did not show
signicant statistical differences from
month to month (f = 0.14, P = 0.84) con-
trary to that of the second group which
showed very signicant statistical differ-
ences (f = 12.99, P = 0.00).
Conclusion: (1) The asthmatic patients,
sensitive to HDM, do not show signicant
uctuation in PEFR during the time, as
long as they receive therapy systematically.
(2) The reported symptoms and the varia-
tion of b2-stimulant-doses show signicant
changes from month to month in patients,
who do not use acaricide products in their
homes. (3) The systematic and correct use
of acaricides results in a steadily improved
clinical situation of the asthmatic patients,
who are sensitive to HDM, throughout to
whole year.
1557
Is yoga training beneficial for exercise
induced bronchoconstriction?
Fulya, T1; Karahancer, S2; Hatice, E3; Kula, E1
1
Erciyes University Hospital, Kaysery, Turkey; 2Karuna
Yoga Center, Kaysery, Turkey; 3Pediatric Allergy
Department, Erciyes University Hospital, Kaysery,
Turkey
Background: Yoga is a scientic system of
physical and mental practices. Preliminary
studies investigating yoga for treating air-
way hyperreactivity have been promising.
This study was undertaken to assess the
benecial effect of yoga in the exercise
induced bronchoconstriction.
Methods: Two groups of children were
enrolled in the study: asthmatic children
with positive (n = 10) and negative
(n = 10) responses to exercise. Both
groups attended one-hour sessions of yoga
training twice a week for three months.
After three months, all children partici-
pated in another exercise challenge.
Results: No signicant difference was
observed in the pre-exercise spirometric
measurements among the groups
(P > 0.05; Mann-Whitney U test). Like-
wise, there was no difference in spirometric
measurements between before and after
yoga training for both groups (P > 0.05,
Willcoxon test.). After yoga training, we
observed signicant improvement in maxi-
mum FEV1 % fall following the exercise
challenge (P < 0.05, Willcoxon test). All
exercise response positive asthmatics
became exercise response negative asthmat-
ics after yoga training.
Conclusion: This study has shown that
practise of yoga training has benecial
effects on exercise induced bronchocon-
striction. It is our opinion that yoga train-
ing can supplement drug therapy in order
to obtain better control of asthma.
1558
The effectiveness of a clinical pathway
for childhood asthma management in an
emergency department
Kim, H1; Chun, Y2; Yoon, J3; Kim, J3; Lee, J3
1
Bucheon St. Marys Hospital, The Catholic University
of Korea, Pediatric Allergy, Pucheon, Republic of Korea;
2
Seoul St. Marys Hospital, The Catholic University of
Korea, Republic of Korea; 3Republic of Korea
Background: This study aimed to deter-
mine whether the clinical pathway based
on evidence is applicable to pediatric
asthma patients in the emergency room
and whether the treatment effectiveness of
this pathway is better.
Method: The asthma clinical pathway was
applied to every child with acute asthma
who visited the emergency room between
Jan 1 and Dec 31, 2009. The emergency
587

room medical records were reviewed to
determine whether if the pathway was cor-
rectly applied. To compare effectiveness,
the patients were divided into 2 groups: the
correct application group and the incorrect
application group.
Result: Of the 51 children with acute
asthma exacerbation, 24 (47%) belonged
to the group in which the clinical pathway
was correctly applied, and 27 (53%)
belonged to the group in which it was
incorrectly applied. Most of the children in
the incorrect application group had moder-
ate asthma exacerbation. Commonly
encountered incorrect applications are no
or insufcient use of inhaled anticholiner-
gics, insufcient use of inhaled bronchodi-
lator, and no use of systemic
corticosteroids. The main cause for the
incorrect application of the clinical path-
way was the inability of the residents to
understand the pathway. The hospitaliza-
tion rate was higher in the patients in
whom critical treatments were incorrectly
applied.
Conclusion: Correct application of the clin-
ical pathway to pediatric asthma patients
in the emergency room increased treatment
effectiveness. However, training for resi-
dents and proper care for patients with
moderate asthma exacerbation are neces-
sary for correct application of the clinical
pathway.
1559
Exhaled nitric oxide fraction in asthmatic
children well correlated with clinical
control of asthma (in primary care)
Badzakova Macheska, G; Obocki, K
Clinical Hospital Sistina-Acibadem, Section for
Allergy and Asthma in Children, Skopje, Macedonia,
FYROM
Background: Fractional exhaled nitric
oxide (FeNO) is a non-invasive marker of
bronchial inammations in asthma.
Numerous studies have conrmed that
FeNO is increased in asthma, increasing
further when asthma control deteriorates,
or when exacerbation occur. FeNO mea-
surement using the hand-held device (Niox
Mino), has been shown to be a valuable
tool in the management of the asthma con-
trol in asthmatic children.
Methods: We are evaluated 116 asthmatic
patients with allergic asthma, for a period
of 6 months. The participating patients
were aged 618 years (mean age 12 years),
wheezes had started before the age of
36 months and they were receiving treat-
ment with inhaled corticosteroid in the
mainly doses of 200500 lg of uticason
dipropionate for at least 12-months dura-
tion. The studied population comprised of
18 uncontrolled children with persistent
588
Poster Session 66 Towards an integrated approach to asthma therapy
wheezy symptoms, 28 partially controlled
and 70 patients with optimal control of
asthma.
Results: 18 asthmatic patient with uncon-
trolled asthma disease (all at teenagers per-
iod of life) have had high-dose of inhaled
corticosteroid treatment and at least one
other asthma controller drug. They all
were allergic poly-sensitization and FeNO
levels were 78.27 (45129) ppb versus par-
tially controlled asthmatic patients 38.5
(1560) ppb. There was a signicant differ-
ence in FeNO levels at the well-controlled
asthmatic children FeNO-15.05 (640) ppb.
(P < 0.001).
Conclusions: Fractional exhaled nitric
oxide has been shown to be a good indica-
tor of asthma control in children. FeNO
levels below 20 ppb were indicators of
good asthma control. The new hand-held
device is enable the introduction of exhaled
NO measurement into the primary health
care.
1560
A comparison of the asthma control test
and spirometry results in the manage-
ment of asthma
Kasternow, B1; Hoda, U2; Alam, S2
1 asthma and 50% of patients had their
Guys and St. Thomas Hospital, London, Adult
Allergy, London, United Kingdom; 2Guys and St.
Thomas Hospital, London, Adult Allergy, London,
United Kingdom
Background: The 2006 Global Initiative
for Asthma (GINA) guideline emphasises
the importance of evaluating asthma con-
trol, rather than asthma severity, in order
to guide asthma management decisions.
Classication of disease severity is a static
measure that, whilst useful in initiating
treatment, is less helpful in guiding subse-
quent treatment. Therefore identifying
parameters that accurately assess control
will benet treatment decisions. The
Asthma Control Test (ACT) is a ve-item
questionnaire for the assessment of asthma
control. An ACT score of <20 has been
shown to correlate with poorly controlled
asthma. Presently, in most respiratory clin-
ics Asthma is classied by severity as mild,
moderate or severe on the basis of
FEV1measurements of > 80%, 6080%
Table 1. For abstract 1560.
ACT >20
N 26
Mean Age 48 (15)
Mean % FEV1 93.7 (26.8)
Physician Judged Controlled 88.4%
Physician Judged Uncontrolled 11.6%
Treatment Increased 3%
Treatment Unchanged 81.6%
Treatment Decreased 15.4%
Allergy 66, Suppl. 94 (2011): 482642  2011 John Wiley & Sons A/S
and <60% of predicted values, respec-
tively. This study compared whether ACTS
or FEV1 correlated better with physicians
assessments of asthma control and treat-
ment decisions made by clinicians in an
outpatient asthma clinic setting.
Method: Serial visits were reviewed to a
specialist Asthma Outpatients Clinic at a
London Teaching Hospital. Patients were
included if they had a diagnosis of asthma
and were able to perform Lung Function
Tests. Patients with other major airways
diseases were excluded. All subjects com-
pleted the Asthma Control Test and per-
formed same day spirometry. Clinicians
made their own assessment of the patients
asthma control and made appropriate
treatment decisions. The clinicians were
not blinded to the results of spirometry or
ACT.
Results: In the group dened as having
controlled asthma (as dened by ACT
>20), 88.4 % of patients were also classi-
ed as controlled by clinicians. However,
in the group with % predicted FEV1
>0.8, only 53.7 % of patients were classi-
ed as controlled by clinicians. In the
group dened as having uncontrolled
asthma (by ACT <20) 75% of patients
were deemed to have poorly controlled
treatment increased. 75.8 % of those
patients were also classied as uncontrolled
by physicians. A very similar percentage
(75.4%) were classied as uncontrolled by
physicians in the group with FEV1 <0.8.
Conclusion: The results of our study show
that an ACT score of <20 had a strong
correlation with the physicians impression
of poor asthma control and correlated
better with treatment decisions than did
the severity of asthma as dened by FEV1.
The ACT could serve as a useful tool in
the assessment of Asthma.
ACT <20 FEV1 >0.8 FEV1 <0.8
96 57 65
48 (14) 48 (14) 47 (14.8)
73.7 (25.7) 103.4 (13.6) 58.1 (16.6)
24.2% 53.6% 24.6%
75.8% 46.4% 75.4%
38.5% 21.4% 42.4%
57.3% 71.5% 54.5%
4.2% 7.1% 6.1%
Poster Session 66 Towards an integrated approach to asthma therapy

1561
The assessments were made by different cant (P < 0.001). According to the criteria
Are assessments of control of asthma investigators to ensure the results free from of GINA, 54.8% of patients were con-
made by the Global Initiative For Asthma bias. In the case of mismatch, the reasons trolled, and 51.0% of all patients had P-
and Pediatric Asthma Control Test com- of discrepancy were investigated. For P- ACT score over 20. There is no concor-
patible with each other? ACT, the core of equal to or higher than dance between decision of control of
20 was assessed as controlled asthma, equal GINA and P-ACT (kappa = 0.464). There
Akan, A; Erkocoglu, M; Civelek, E; Azkur, D; Ozcan, C;
Kaya, A; Kocabas, C
to and lower than 19 as uncontrolled. And were no difference between the patients
Ankara Hematology Oncology Childrens Research and standard GINA criteria were considered. classied as controlled asthma with both
Training Hospital, Pediatric Allergy and Immunology, Result: 314 children with asthma were methods and the others classied as par-
Ankara, Turkey
enrolled to the study. 56.1% (n = 176) of tially controlled and uncontrolled with both
them were male and the mean age was of the methods according to age, gender,
Background: Pediatric Asthma Control
9.0 0.1 (mean SD error). The P-ACT age at diagnose, severity of asthma, dura-
Test (P-ACT), is a widely used and recom-
score for all of the patients was 20.0 tion of asthma, time of follow-up.
mended tool for the follow-up of child-
(15.023.0) (median [2575%]); the ratio of Conclusion: It is elucidated that there is no
hood asthma. The results of P-ACT are
controlled, partially controlled and uncon- concordance about the assessment of con-
usually compatible with GINA, but infre-
trolled patients were 54.8%, 29.9%, trolled or uncontrolled asthma between
quently they can be discordant with each
15.3%, respectively. The differences GINA and P-ACT. Larger studies with dif-
other in daily practice.
between P-ACT scores of patients classied ferent patient groups are necessarily needed
Method: Firstly, the grade of consistency
as controlled, partially controlled and to establish the factors causing discrepancy
between the assessment of control made by
uncontrolled based on the criteria of between these two methods.
doctor using GINA guidelines and the
GINA are found to be statistically signi-
patient with P-ACT survey was searched.

Allergy 66, Suppl. 94 (2011): 482642  2011 John Wiley & Sons A/S 589
 Poster Session 67

Occupational allergy

1562 Table 1. For abstract 1562. Distribution of the studied cases by their factory products
Work related asthma in the Damietta Asthmatic Asthmatic Nonasthmatic Nonasthmatic Total Total
Factory
governorate
product Number % Number % Number % P value
Elshabrawi Ibrahim, A1; Abu Najlah, A2; Sedky, M2;
Elnawawy, A3; Montaser, M4 Wood 47 8.1 535 91.9 582 54.7 0.001
1
Damiietta Faculty of Medicine, Chest Diseases Marble 8 8.5 86 91.5 94 8.8 0.001
Department, Al-Azhar University, New Damietta City, Textile 14 8.75 146 91.25 160 15.03 0.001
Damietta Governorate, Egypt; 2Faculty of Medicine, Cotton 12 16.2 62 83.8 74 6.9 0.001
Chest Diseases Department, Al-Azhar University (Cairo),
Plastic 4 4.4 87 95.6 91 8.6 0.001
Cairo, Egypt; 3Damiietta Faculty of Medicine, Public
Health and Community Medicine Department, Al-Azhar
Pottery 5 7.9 58 92.1 63 5.9 0.001
University, New Damietta City, Damietta Governorate, Total 90 8.5 974 91.5 1064 100 0.001
Egypt; 4Chest Diseases Department, Al-Azhar University
Hospital in Damietta, New Damietta City, Damietta
Governorate, Egypt Table 2. For abstract 1562. Distribution of the studied cases by their factory products
and work type
Background: The association between
Asthmatic Asthmatic Nonasthmatic Nonasthmatic
asthma and occupation is recognized since
antiquity. Work related asthma refers to Number % Number % X2 P value
all situations where asthma is inuenced by Factory product
work and encompasses both occupational Wood 47 52.2 535 54.9 FE 0.66
asthma (OA) and work exacerbated Nonwood 43 47.8 439 45.1 FE 0.66
asthma (WEA). This study investigated Work type
some occupational exposures and their Ofce work 3 3.3 97 10 4.89 0.08
Machine work 82 91.1 806 82.8 4.89 0.08
relations to work related asthma in Dami-
Cleaning work 5 5.6 71 7.3 4.89 0.08
etta Governorate.
Total 90 100 974 100
Method: Workers in Furniture, Textiles,
Plastic, Pottery and Marble factories in the
industrial zone in New Damietta City, Table 3. For abstract 1562. Relation between factory products and skin prick tests
were the target sample for this study. Cross Factory product (90)
sectional study was conducted to investi-
gate asthmatic workers in Damietta gover- Wood (47) Wood (47) Non Wood (43) Non Wood (43)
norate, Self administered questionnaire, Number % Number % X2 P value
pulmonary function tests before and after Wood 21 44.7 3 7 16.3 0.000
inhaled bronchodilators, PEFR at start Mites 4 8.5 4 9.3 FE 1.000
and end of working shifts, serum IgE., Tobacco 17 36.2 14 32.6 FE 0.83
Sputum and blood eosinophilic count and Wheat 7 14.9 5 11.6 FE 0.76
skin prick testing were done to workers Cotton dust 4 8.5 14 32.6 8.2 0.004
involved in this study. House dust 19 40.4 12 27.9 FE 0.27
Result: The prevalence of occupational Hay 7 14.9 5 11.6 FE 0.76
Banana 9 19.1 4 9.3 FE 0.24
asthma in studied populations was 8.45%
Strawberry 4 8.5 6 14 FE 0.51
(90/1064). The major affected age group
Fish 5 10.6 9 20.9 FE 0.25
was from 35 to 54 (52.3%); males were
more prevalent (90.2%); Wood factories
were the most common in the present work
(45.7%). pational rhinitis have close relationships.
1563
Conclusion: WRA could be efciently diag- The aim of the present study was to inves-
Abstract withdrawn
nosed questionnaire based and using peak tigate the prevalence of occupational rhini-
ow meters, pre-employment and periodic tis in subjects with occupational asthma,
medical check-up help to pick up cases of 1564 according to the causal agents, and the
WRA early. Physicians recognizing new Occupational rhinitis in workers with temporal relationships between the two dis-
cases of OA at a work site should suggest occupational asthma eases.
introduction of environmental controls and Methods: New cases of occupational
Ameille, J1; Hamelin, K1; RNV3P, M2
an action plan to treat these patients. 1
Institut interuniversitaire de medecine du travail, Paris, asthma (20082009) were collected by a
France; 2ANSES, Maisons Alfort, France national surveillance programme, based on
voluntary reporting, involving a network
Background: Increasing scientic evidence of physicians working in occupational
shows that occupational asthma and occu- medicine departments in French university
590 Allergy 66, Suppl. 94 (2011): 482642  2011 John Wiley & Sons A/S
Poster Session 67 Occupational allergy
hospitals (Reseau National de Vigilance et
de Prevention des Pathologies Professionn-
elles, RNV3P). The participating physi-
cians were asked to report all new cases of
occupational asthma that they investigated,
using a reporting from including informa-
tion on age, sex, occupation at the time of
diagnosis, suspected causal agent, presence
or absence of rhinitis symptoms, and
respective dates of occurrence of rhinitis
and asthma.
Results: Four hundred and thirty ve cases
of occupational asthma were reported in
20082009. Mean age was 40.1 11.6
years. The greatest numbers of cases were
observed in bakers, hairdressers, cleaners
and health workers (15.4%, 13.1%, 13.7%,
10.7% of cases, respectively). The causal
agents most frequently incriminated were
our, quaternary ammoniums, persulfate
salts, and isocyanates (15.6%, 12.2%,
12.2% and 9.4% of cases, respectively).
Rhinitis was associated with asthma in 245
cases (56.3%). Symptoms of rhinitis
occurred before symptoms of asthma in
48% of cases, contemporaneously with
asthma in 35% of cases, at a later date in
8% of cases (undetermined: 9% of cases).
Symptoms or rhinitis were more prevalent
in cases of occupational asthma attributed
to high molecular weight agents, than in
cases attributed to low molecular weight
agents (73% versus 48%, P < 0.05).
Conclusion: These results conrm the fre-
quent association between occupational
asthma and occupational rhinitis and sup-
port the united airways disease concept.
1565
Prevalence of chlorhexidine specific IgE
sensitisation in healthcare workers
attending an occupational health depart-
ment
Alam, S1; Thayalan, S1; Nagendran, V2
1
Occupational Health Department, St.Georges Hospital,
London, United Kingdom; 2Department of Immunology
& Allergy, Epsom & St. Helier University Hospitals NHS
Trust, Carshalton, United Kingdom
Background: Chlorhexidine is an antiseptic
and disinfectant which is widely used in
hospitals as handwashing agents (Hibi-
scrub, Hydrex, Hibisol). In the UK, recent
campaigns to eradicate hospital acquired
infections has increased the use of these
agents, and we now have guidelines recom-
mending the use of 2% chlorhexidine glu-
conate in 70% isopropyl alcohol solution
for skin decontamination before insertion
of central venous cannulae, and for cleans-
ing when dressings for these devices are
changed. The allergenic potential of
chlorhexidine in health care workers
(HCWs) was highlighted for the rst time
in 2008. Our objective was to investigate
Allergy 66, Suppl. 94 (2011): 482642  2011 John Wiley & Sons A/S
the extent of this problem in HCWs
attending an occupational health depart-
ment in a large tertiary care hospital in
London, UK.
Method: A cross sectional prevalence study
was conducted to estimate IgE sensitisation
to chlorhexidine and to examine the risk
factors associated with chlorhexidine sensi-
tivity. HCWs attending the occupational
health department were invited to partici-
pate in the study. Chlorhexidine exposure,
frequency of handwashing, medical history
and occupational history were obtained
using a questionnaire, and chlorhexidine
specic IgE was measured in a blood sam-
ple (by Phadia ImmunoCAP method).
Result: A total of 225 healthcare workers
(63% females, 37% males; age range 18
62) participated in the study. 133 of these
HCWs reported previous occupational
exposure to chlorhexidine, and 4 out of
this group (3 females, 1 male) were chlorh-
exidine specic IgE positive, giving a prev-
alence rate of 3%. Three of the 4 positive
cases belonged to the group with a high
frequency of handwashing. There were no
chlorhexidine positive results in the 92
HCWs who were new entrants to health
care work, with no previous occupational
exposure to Chlorhexidine.
Conclusion: This study conrms the impor-
tance of chlorhexidine as a potential occu-
pational hazard for healthcare workers. As
with latex allergy, chlorhexidine allergy
carries a potential risk of anaphylaxis.
With the increasing use of chlorhexidine
containing handwashing agents in the
healthcare setting, it is important to test
Specic IgE to Chlorhexidine when a
HCW presents with allergic symptoms.
1566
Occupational asthma - 13 years experi-
ence in the Czech Republic
Balkova, B; Lebedova, J; Klusackova, P; Hurtova, L;
Fenclova, Z
Department of Occupational Medicine, General
University Hospital, Prague 2, Czech Republic
Background: To provide information about
occupational asthma (OA) over the last
13 years with a country-wide population of
approximately 10 million.
Method: Three hundred and ninety eight
patients presented with possible OA at the
Department of Occupational Medicine of
General University Hospital in Prague
between years 19962009 and undertook
examination. For conrmation of OA posi-
tive histamine/methacholine challenges and
positive specic bronchoprovocation tests
were essential. A decline in forced expiratory
volume in one second parameter (FEV1) of
20 or more percent from the baseline reading
was considered as positive test.
Result: Throughout Czech Republic OA
was reported in 678 cases (3.4% of all noti-
ed occupational diseases) during analyzed
period. The most frequent allergens were:
low-molecular-weight (LMW) - disinfec-
tant, isocyanates, high-molecular-weight
(HMW) wheat and rye ower. Within
group of patients examined at our depart-
ment, OA was fully diagnosed in 23.4%.
The average age of subjects was 39.4, the
mean exposition time was 111 months, and
the duration of difculties till the conrma-
tion of occupational disease was
20 months. Patients were examined on
average 5.7 months after leaving the work-
place. Within group of patients exposed to
HMW allergens the duration of exposition
and duration of health difculties were
longer than in those exposed to LMW.
The difference was statistically signicant
(P < 0.05).
Conclusion: Since 1996 the annual numbers
of new OA diagnoses have been relatively
constant (in average 50 per year). OA is
relatively rare occupational disease how-
ever it seems to be under-diagnosed. OA
should be considered in daily clinical prac-
tice by all physicians. In adults, patients in
whom an occupational association is sus-
pected should be referred for specic
assessments.
1567
Spontaneous pneumomediastinum: an
uncommon complication of a common
disease
Pala, G1; Pignatti, P2; Moscato, G2
1
Salvatore Maugeri Foundation - Scientific Institute of
Pavia, Allergy and Clinical Immunology Unit, Pavia,
Italy; 2Italy
Background: Spontaneous pneumomediast-
inum (SPM) is a benign condition, often
presenting in young adults exposed to a
sudden pressure change within the intra-
thoracic cavity. A history of asthma has
been reported as a factor in the develop-
ment of SPM in up to 50% of cases.
Approximately 10 to 15% of cases of adult
asthma are attributable to occupational
factors. Although the long term conse-
quences of work-related-asthma are well
known, acute complication, but not SPM,
has rarely been described.
Case description: We report on a 24-year-
old baker with a history of rhinoconjuncti-
vitis, cough, shortness of breath and
wheezing occurring when he was exposed
to wheat our at the workplace. He con-
trolled symptoms by daily use of short-act-
ing beta-agonists as relief medication, was
never submitted to medical surveillance at
the workplace, and never consulted a spe-
cialist. While working on Christmas Eves
night, when bread production is increased,
591

he started to complain of progressively
worsening shortness of breath and cough
inducing vomiting and, few hours later,
chest pain, and he was admitted to the
Emergency Department. Chest radiography
and subsequent CT scan revealed pneumo-
mediastinum.
Results: When he came to our observation,
nine months after the acute event, baseline
spirometry was normal. Fraction of
exhaled nitric oxide was elevated (52 ppb)
and methacholine challenge test showed
marked bronchial hyperresponsiveness
(PD20FEV1: 206 lg). Skin prick tests were
positive for wheat our extract (wheal size
15 8 mm) and specic serum IgE to
wheat our were 5.79 kU/l. The specic
inhalation challenge was not performed
due to the previous history of SPM, but
peak expiratory ow monitoring showed
signicant variations related to workplace
exposure. A diagnosis of occupational
asthma due to wheat our was made.
Conclusions: Our case highlights that in
workers exposed to occupational allergens
the lack of medical surveillance or special-
ist consultation, and subsequently the delay
in diagnosis and treatment of work related
asthma may provoke acute complications,
such as SPM, enhancing the importance of
prevention.
1568
Prevalence and determinants of asthma
and rhinitis in an Italian cleaning
company from Terni, Italy
Folletti, I1; Bussetti, A2; Murgia, N3; Profeta, N3;
Siracusa, A1
1
University of Perugia, Clinical and Experimental
Medicine, Terni, Italy; 2Narni, Italy; 3University of
Perugia, Clinical and Experimental Medicine, Perugia,
Italy
Background: Professional and domestic
cleaning has been associated with new
onset and work-exacerbated asthma and
rhinitis. Cleaners are exposed to multiple
cleaning agents. The aim of this study was
to assess the prevalence and determinants
of asthma and rhinitis in 234 subjects
working in an Italian cleaning company
based in Terni.
Method: In this cross-sectional study a
cohort of 234 cleaning company workers
(126 cleaners and 108 non-exposed con-
trols), completed the Italian version of the
ECRHS I and ECRHS II occupational
modules to assess exposure to cleaning
agents and prevalence of asthma and rhini-
tis. They also performed lung function
tests. Statistical differences in continuous
variables were tested by the two-tailed
paired t test. Differences in proportions
were tested by the V2 or Fishers exact test,
when appropriate. Differences were consid-
592
ered signicant at P < 0.05. The inuence
of cleaning tasks or products, schooling,
sex, age and smoking status (predictors) on
asthma or rhinitis (dependent variables)
was analyzed by logistic regression.
Result: The 234 subjects had a mean age
of 44 years; cleaners were 4 years older
than controls. 79% were female and 43%
were current smokers. Median schooling
was 17 years (range 1024). 54% of the
cohort worked as cleaners in public build-
ings, hospitals and schools, and 46% were
white-collar workers and drivers who were
not exposed to cleaning products (con-
trols). The prevalence of current asthma
was 6% in cleaners and 1% in controls.
Prevalence of rhinitis was 17% and 15%,
respectively. Using glass cleaning sprays at
work 1 day/week was associated with cur-
rent asthma (OR = 19.0, CI 2.1159.9).
Cleaners <39 years old had a greater risk
of asthma (OR = 5.0, CI 1.221.7) than
older co-workers, after adjusting for other
predictors.
Conclusion: We conrm that exposure to
cleaning products is associated with
asthma. Use of glass cleaning sprays is an
important determinant. The healthy
worker effect could account for the higher
risk of asthma that we found in younger
cleaners. Moreover, there is evidence that
the incidence of work-related symptoms is
higher in the rst 23 years after starting
exposure and tends to decrease thereafter.
1569
Determinants of baseline and serial frac-
tional exhaled nitric oxide levels in spice
mill workers with work-related respira-
tory allergy and asthma
Van der Walt, A1; Singh, T2; Baatjies, R1; Jeebhay, M1
1
School of Public Health and Family Medicine, Centre
for Occupational and Environmental Health Research,
University of Cape Town, Cape Town, South Africa;
2
National Institute for Occupational Health,
Immunology, Johannesburg, South Africa
Background: Measurement of fractional
exhaled nitric oxide (FeNO) is suggested as
an important non-invasive marker for air-
way inammation in asthmatics. However,
various occupational studies have demon-
strated inconsistent results. This study eval-
uated the determinants of elevated FeNO
(>50 ppb) for clinically relevant allergic
respiratory disease endpoints in spice mill
workers.
Method: A cross-sectional epidemiological
study of 150 workers used an interviewer-
administered ECRHS questionnaire
adapted for local conditions. IgE reactivity
to common inhalants (Phadiatop) and
occupational allergens (garlic, chili pepper
and wheat) (Phadia, ImmunoCAP) was
quantied. Spirometry and FeNO were
Allergy 66, Suppl. 94 (2011): 482642  2011 John Wiley & Sons A/S
Poster Session 67 Occupational allergy
determined using ATS/ERS criteria (2005).
A hand-held portable nitric oxide sampling
device (NIOX MINO, Aerocrine AB)
measured FeNO before and after the 8-
hour work shift and 24 h prior to the next
shift.
Result: The mean age of workers was
33 years, 71% were male, 46% current
smokers and 45% atopic. Spice dust
related ocular-nasal symptoms (43%) were
more common than asthma symptoms
(17%). Sensitisation to garlic (19%) was
higher than to wheat (9%) or chilli pepper
(6%). There were 8% of workers with ele-
vated FeNO (>50 ppb) suggestive of
asthma. The baseline pre-shift FeNO geo-
metric mean (GM 14.9 ppb) was very simi-
lar to the mean change across shift (GM
15.4 ppb) and across the 24-hour period
(GM 15.8 ppb). A lower proportion had a
FeNO increase more than 60% across shift
(3%) than across the 24-hour period from
baseline (5%). Atopy and smoking equally
explained most of the variability
(r2 = 0.09, P < 0.001) in baseline pre-shift
FeNO. Sensitisation to chilli pepper
(r = 0.32) was more strongly correlated
with FeNO than garlic (r = 0.20). Chilli
pepper persisted as a strong determinant
(b = 0.47) of FeNO similar to smoking
(b = )0.47), but less so than atopy
(b = 0.41) and recent green vegetable
intake (b = 0.28) in the multivariate linear
model. Probable asthma (FeNO >50 ppb)
was also more strongly associated with
chilli pepper sensitisation (OR 23.9, CI
5.2109.2) than wheat (OR 6.4, CI 1.6
25.0) or garlic (OR 5, CI 1.516.9). None
of the variables were associated with an
increase in FeNO (>60%) across shift or
over 24 h.
Conclusion: Sensitisation to occupational
allergens, chilli pepper and to a lesser
extent garlic, is a major determinant of ele-
vated levels of FeNO in spice mill workers.
Current smoking and atopy modulate the
FeNO response in opposite directions.
1570
Respiratory symptoms and lung function
among chemical workers
Stoleski, S1; Karadzinska-Bislimovska, J1; Minov, J1;
Mijakoski, D2; Risteska-Kuc, S2; Marsenic, M2; Trajceva,
L3
1
Department for Cardiorespiratory Functional
Diagnostics, WHO Collaborating Center, GA2LEN
Collaborating Center, Institute for Occupational Health
of R. Macedonia, Skopje, Macedonia, FYROM; 2WHO
Collaborating Center, GA2LEN Collaborating Center,
Allergy Center, Institute for Occupational Health of R.
Macedonia, Skopje, Macedonia, FYROM; 3WHO
Collaborating Center, GA2LEN Collaborating Center,
Institute for Occupational Health of R. Macedonia, ENT,
Skopje, Macedonia, FYROM
Aim of the study: The aim of this study is
to evaluate the prevalence of respiratory
Poster Session 67 Occupational allergy
symptoms in chemical workers and to
assess the effect of workplace exposure on
ventilatory capacity, lung functional
impairment, and bronchial hyperrespon-
siveness (BHR).
Methods: We performed a cross-sectional
study investigating a group of 45 female
chemical workers aged 2159 years. In
addition, 43 female ofce workers aged 22
to 58 years were examined as a control.
Evaluation of examined subjects included
completion of a questionnaire on respira-
tory symptoms in the last 12 months
(cough, phlegm, dyspnea, wheezing, and
chest tightness). Lung function was mea-
sured by recording maximum expiratory
ow-volume (MEFV) curves. Forced vital
capacity (FVC), forced expiratory volume
in the rst second (FEV1), forced expira-
tory ratio (FEV1/FVC%), and ow rates
at 75%, 50% and the last 25% of the vital
capacity (MEF75, MEF50, MEF25) were
measured on the MEFV curves. BHR was
evaluated by histamine challenge
(PC20 8 mg/ml) in both groups.
Results: The data analysis showed a signi-
cantly higher prevalence of overall respira-
tory symptoms in chemical workers than in
controls (34.5% versus 12.7%, P < 0.05),
being the highest for phlegm (19.3% versus
4.1%, P < 0.01), dyspnea (15.7% versus
5.8%, P < 0.05), and wheezing (14.3%
versus 6.1%, P < 0.05). All spirometric
parameters (FVC, FEV1, FEV1/FVC%,
MEF75, MEF50, and MEF25) were signif-
icantly lower in chemical workers. Preva-
lence of BHR was higher in chemical
workers but without statistical signicance
(19.6% versus 12.8%, P > 0.05). Signi-
cantly higher prevalence was registered for
borderline BHR in chemical workers
(9.8% versus 2.9%, P < 0.05). Duration
of exposure to chemicals was not related
with the decline in lung function tests,
whereas smokers had lung function
changes similar to those of non-smokers,
indicating that smoking habit did not
account for all respiratory effects.
Conclusion: Our data suggest signicant
adverse respiratory effects of occupational
exposure among chemical workers associ-
ated with the development of chronic respi-
ratory symptoms and impaired lung
function.
Allergy 66, Suppl. 94 (2011): 482642  2011 John Wiley & Sons A/S
1571
Domestic mite antigens in floor and air-
borne dust at workplaces in comparison
to living areas
Sander, I1; Zahradnik, E1; Kraus, G2; Mayer, S3;
Neumann, H4; Fleischer, C1; Bruning, T1; Raulf-
Heimsoth, M1
1
IPA - Institute for Prevention and Occupational
Medicine DGUV, Ruhr-Universitat Bochum, Bochum,
Germany; 2Berufsgenossenschaft Energie Textil Elektro
Medienerzeugnisse (BGETEM), Augsburg, Germany;
3
Berufsgenossenschaft Handel und Warendistribution
(BGHW), Mannheim, Germany; 4Unfallkasse Nordrhein-
Westfalen, Dusseldorf, Germany
Background: Allergens produced by
domestic mites (DM) are among the most
common triggers of asthma. While house
dust mite allergen concentration in oor or
mattress dust of living areas is well studied,
there is a lack of data concerning airborne
mite allergen exposure. It was the aim of
the study to compare occupational mite
antigen exposure with living areas. Thus,
besides oor dust personal inhalable dust
samples were analysed.
Method: At workplaces and in living areas
203 oor dust samples and 87 personal in-
halable dust samples were collected. For
sensitive quantication of DM antigens a
new enzyme immunoassay (EIA) based on
polyclonal antibodies (pAb) to Dermato-
phagoides farinae extract was developed.
Its reactivity to other astigmata was
checked with extracts from eight mite spe-
cies. All dust samples were tested with the
new EIA and, in addition, with the:Der f
1 EIA based on monoclonal antibodies
(Indoor Biotechnologies, UK). Samples
below the detection limit in the DM-EIA
were retested in an assay variant with a u-
orogenic substrate (DM-FEIA).
Result: The newly developed DM-EIA
does not react with cereals, seeds and
moulds, but shows cross-reactivity to the
extracts from other astigmata. The DM-
EIA has a lower detection limit to D. fari-
nae protein of 200 pg/ml, and the DM-
FEIA of 50 pg/ml. DM antigens were
detected by DM-EIA/FEIA in all oor
dusts and 75 (86%) airborne samples,
while Der f 1 was found in 123 (61%) oor
dust and 5 airborne samples only. Even so,
Der f 1 and DM concentrations were
highly correlated (Pearson r = 0.98, Pear-
son of log-values r = 0.86, P < 0.0001).
DM-antigen levels were signicantly higher
in personal airborne samples from textile
recycling (median 288 ng/m3, n = 7), bed
feather lling (37 ng/m3, n = 8), feed pro-
duction (36 ng/m3, n = 4), grain storage
(27 ng/m3, n = 5) and cattle stables (9 ng/
m3, n n = 17) in comparison to living
areas (2 ng/m3, n = 16). In oor dust sam-
ples only values from textile recycling were
signicantly higher than from living areas
while several other workplaces (e.g.
schools, ofces) showed lower values.
Conclusion: As EIAs specic to single
house dust mite allergens are not feasible
for measuring personal airborne exposure,
a new pAb based EIA directed to DM
antigens was developed. DM antigen levels
are well correlated to Der f 1 values, and
were measurable in 86% of personal inha-
lable dust samples. At some workplaces
signicantly higher DM antigen exposure
than in living areas occurred.
1572
Nasal symptoms in textile workers: rela-
tion to respiratory symptoms, atopy,
bronchial hyperresponsiveness and dura-
tion of exposure
Mijakoski, D1; Karadzinska-Bislimovska, J2; Stoleski, S2;
Minov, J2; Marsenic, M1; Atanasovska, A1
1
Institute for Occupational Health of RM, WHO CC,
GA2LEN CC, Allergy Center, Skopje, Macedonia,
FYROM; 2Institute for Occupational Health of RM, WHO
CC, GA2LEN CC, Department for cardiorespiratory
functional diagnostics, Skopje, Macedonia, FYROM
Background: Objective: To determine if
nasal symptoms were associated with respi-
ratory symptoms in textile workers, and to
assess their relation to atopy, bronchial
hyperresponsiveness (BHR) and duration
of exposure.
Method: We performed a cross-sectional
study including 52 textile workers, 15 males
and 37 females, mean age 32.5 4.5 years,
mean duration of exposure 15.5
4.8 years. In addition, 56 ofce workers (17
males and 39 females, mean age 30.4
5.3 years) were examined as a control. Eval-
uation of examined subjects included com-
pletion of a questionnaire on nasal and
respiratory symptoms in the last 12 months,
skin prick tests to common inhalant aller-
gens and workplace allergens (cotton, wool,
silk, synthetics), spirometry, and histamine
challenge test (PC20 8 mg/ml).
Result: Prevalence of overall nasal symp-
toms in the last 12 months was signi-
cantly higher in textile workers (36.5%
versus 14.3%, P < 0.05). The difference
was statistically signicant for sneezing
(36.5% versus 12.5%, P < 0.05) and rhin-
orrhea (38.5% versus 14.3%, P < 0.05).
Nasal symptoms were signicantly associ-
ated with respiratory symptoms and atopy
in both examined groups. Signicant asso-
ciation was registered between nasal symp-
toms and BHR only in textile workers
(P < 0.05). The prevalence of nasal symp-
toms in textile workers was signicantly
higher in the subjects with workplace expo-
sure longer than 15 years (P < 0.05).
Conclusion: Our data indicate high preva-
lence of nasal symptoms in textile workers
and their signicant relation to respiratory
593

symptoms, atopy, BHR and to duration of
occupational exposure.
1573
Does atopy predispose to asthma exacer-
bations at work? A study on predictors
of work exacerbated asthma
Murgia, N; Paolocci, G; Gambelunghe, A; Palumbo, G;
dellOmo, M; Muzi, G; Abbritti, G
Section of Occupational Medicine, Respiratory Diseases
and Toxicology, University of Perugia, Perugia, Italy
Background: Asthma represents a consider-
able source of morbidity and economic bur-
den. Workplace exposures can cause
occupational asthma or trigger a non-occu-
pational asthma, conguring a work exac-
erbated asthma (WEA). Recent data have
conrmed that WEA has a similar socio-
economical impact of occupational asthma.
Little is known about the inuence of atopy
in the development of WEA. The aim of
this study is to investigate whether atopy or
other factors are predictors of WEA in a
group of asthmatics in working age.
Method: 342 subjects in working age with a
diagnosis of asthma, based on GINA guide-
lines and a positive methacholine test (or a
signicant bronchodilator response), were
interviewed to assess the presence of WEA
and to evaluate factors associated with work
related asthma exacerbations. All the sub-
jects underwent a spirometry and skin prick
tests for common aeroallergens. Patients
with occupational asthma were excluded.
Result: 38.3% of the population reported
WEA (n = 131); 80% of the patients with
WEA and 80.2% of the other asthmatics
shown hypersensitivity to seasonal and/or
perennial allergens. Subjects with work
exacerbated asthma have a more severe
asthma in terms of rescue medications use
(OR 1.02, CI 95% 1.011.03) and uncon-
trolled respiratory symptoms in the last
12 months (OR 2.23; CI 95% 1.383.77).
Atopy and hypersensitivity to seasonal
and/or perennial allergens were not risk
factors for developing WEA. Occupational
exposure to vapours, dust, gas and fumes
(VGDF) was correlated with the occurrence
of WEA (OR 3.03; CI 95% 1.544.68).
Conclusion: In this study atopy did not
shown any association with work exacer-
bated asthma, while occupational exposure
to known asthmogens, as VGDF, seemed to
play an important role in workplace asthma
exacerbations. This nding, despite the
small number of the population and the
high prevalence of atopy, could suggests
that pre-employment selection based on
atopy is not effective in preventing WEA,
whereas more attention has to be paid to
reduce workplace exposure to asthmogens.
594
1574
Risk factors associated with work-related
asthma phenotypes in dental health care
workers
Singh, T1; Bello, B2; Jeebhay, M3
1
National Institute for Occupational Health, NHLS,
Immunology, Johannesburg, South Africa; 2National
Institute for Occupational Health, NHLS, Epidemiology,
Johannesburg, South Africa; 3Centre for Occupational
and Environmental Health Research, School of Public
Health and Family Medicine, University of Cape Town,
Cape Town, South Africa
Background: Exposure in the dental envi-
ronment has been shown to increase the risk
of respiratory disease in dental health care
workers (HCWs). This study investigated
the prevalence of work-related asthma phe-
notypes in dental HCWs and the risk fac-
tors associated with these phenotypes.
Method: A cross sectional study was con-
ducted of 454 dental HCWs including den-
tal students in ve academic dental
institutions in South Africa. A self admin-
istered modied ECRHS questionnaire
eliciting the health and employment history
was used. Sera was analysed for atopic sta-
tus (Phadiatop). Pre and post bronchodila-
tor spirometry was conducted using ATS/
ERS guidelines (2005). Multiple logistic
regression analysis was conducted to assess
the association between risk factors and
asthma phenotypes controlling for known
confounders (age, smoking, gender).
Result: The study demonstrated a 6.9%
prevalence of atopic asthma, 5.9% for
non-atopic asthma and 4.0% for work-
aggravated asthma (WAA). Almost half
(46%) of the study population was atopic
and this factor was a signicant predictor
of WAA (OR: 3.4; 95% CI: 1.110.8). A
history of ocular-nasal symptoms was sig-
nicantly associated with WAA (OR: 5.9;
95% CI: 1.917.8). Similarly, work-related
ocular-nasal symptoms were more strongly
associated with WAA (OR: 6.7; 95% CI:
2.419.1). Workers with non-atopic asthma
were more likely (OR: 3.0; 95% CI: 1.2
7.4) to have a longer employment duration
(>20 years).
Conclusion: Dental HCWs with ocular-
nasal symptoms and atopic individuals are
at increased risk of presenting with work-
aggravated asthma. Identication of vari-
ous asthma phenotypes can provide a use-
ful tool for ongoing surveillance and
clinical management of dental HCWs with
asthma symptoms.
Allergy 66, Suppl. 94 (2011): 482642  2011 John Wiley & Sons A/S
Poster Session 67 Occupational allergy
1575
Occupational allergic symptoms in a
chewing gum factory: working place con-
ditions are important
Ozseker, F1; Tepetam, F1; Gelincik, A2; Ozmen, I3;
Calisir, H3
1
HM Sureyyapasa Chest Diseases and Chest Surgery
Training Hospital, Allergy and Immunology, Istanbul,
Turkey; 2Istanbul University Istanbul Medical Faculty,
Allergy and Immunology, Istanbul, Turkey; 3HM
Sureyyapasa Chest Diseases and Chest Surgery
Training Hospital, Chest Diseases, Istanbul, Turkey
Background: Food workers are exposed to
different organic and inorganic dusts. Any
of these chemicals can result in allergic
reactions in sensitized individuals. We pres-
ent multiple cases of occupational allergy
in the same chewing gum factory.
Method: A majority of workers (17 of 22,
F/M = 2/15, mean age: 30, 64 6.12
years) in a special section of a chewing
gum factory had begun to complain from
respiratory and skin symptoms after they
have started to product a new kind of
gum. In all these patients, allergy skin
prick tests were performed with inhalant
allergen pannel (ALK, Albello, Denmark)
and all raw materials (15) being used in
gum production. Patch tests were per-
formed with raw materials in two workers.
In 58 healthy controls skin prick tests were
performed with 15 raw materials. In addi-
tion, the factory was visited to evaluate the
conditions in the production unit. Most of
the production stages were taken place in
uncovered containers and belts.
Result: The mean duration from the begin-
ning of the new production until the appear-
ances of symptoms was 24 11 days. The
skin prick tests with gum Arabic, menthol
and eucalyptus were positive (4/8) in four
patients, menthol and cool boost in one
patient (1/8), gum Arabic and menthol in
one patient (in same patient cereal pollens
allergy was present) (1/8), in one patient
with gum Arabic, peppermint and eucalyp-
tus (1/8), in the other patient only with gum
Arabic (1/8). Laryngeal edema in one
patient, and syncope, bradycardia with ar-
rythmia in another patient occurred ve
minutes after the skin prick tests were
applied. Of all healthy controls prick tests
were negative. Patch tests were found nega-
tive in two patients. After the workplace air-
conditioning system was reconstructed and
the belts were covered, all symptoms of
workers disappeared.
Conclusion: Although rare, Type I allergic
reactions have been described with most of
the above mentioned ingredients. An aller-
gic reaction due to these chemicals must be
taken into account in case of unexpected
symptoms in exposed subjects. However,
disappearance of the symptoms in all the
patients after the workplace conditions
were improved displays that diminishing
Poster Session 67 Occupational allergy
the exposure rate with the possible sensi-
tizer is of utmost importance.
1576
Allergic symptoms in a large group of
young students
Arcangeli, G; Montalti, M; Piccioli, A; Pristera, L; Mucci,
N; Cupelli, V
Department of Public Health - Section of Occupational
Medicine, University of Florence, Florence, Italy
Background: The aim of the study is to
evaluate the incidence of referred allergic
symptoms in a population of young people
living in Tuscany, Italy.
Method: We enrolled 1038 students (439
males and 599 females) of professional
schools (catering, dental technicians, dieti-
tians, obstetrics and laboratory technicians
schools): 188 (107 males and 81 females)
people out of them were younger than
16 years (Group I), 331 (165 males and
166 females) were 1618 years old (Group
II), 213 (69 males and 213 females) were
1821 years old (Group III) and 306 (98
males and 208 females) were older than
21 years (Group IV). Two questionnaires
were administered by resident physicians:
the rst to investigate respiratory issues
(with our items and other items from EC-
RHS I/II and ISAAC questionnaires), the
other one to investigate dermatological
issues (the Italian version of the NOSQ
questionnaire). Statistical analysis was
done using open source software.
Result: In the whole population the inci-
dence of positive cases was 24.6% (males
25.9%, females 23.5%) for oculorhinitis;
15.1% (males 17.1%, females 13.7%) for
asthma; 12.9% (males 12.5%, females
13.2%) for hand dermatitis and 13.0% for
urticaria (males 12.3%, females 13.5%).
The incidences in groups, from I to IV,
were respectively: 26.6%, 25.0%, 26.3%,
21.6% for oculorhinitis; 17.0%, 17.5%,
18.3%, 9.2% for asthma (P < 0.05);
13.3%, 12.4%, 16.0%, 11.1% for hand
dermatitis and 16.0%, 14.5%, 10.8%,
11.1% for urticaria (P < 0.05). In the rst
group the prevalence of hand eczema and
oculorhinitis was higher in males than in
females (P < 0.05).
Conclusion: The prevalence of oculorhinitis
seems to be stable during the growing age.
The incidence of hand dermatitis, urticaria
and asthma appears to decrease, particu-
larly after the age of 21. Among the youn-
gest population group males have referred
a signicantly higher incidence of hand
eczema and oculorhinitis than females.
Allergy 66, Suppl. 94 (2011): 482642  2011 John Wiley & Sons A/S
1577
Work-related respiratory disease and
asthma in spice mill workers is associ-
ated with inhalant chili pepper and garlic
exposures and allergic sensitisation
Jeebhay, M1; Van der Walt, A1; Singh, T2; Baatjies, R1;
Lopata, A3
1
School of Public Health and Family Medicine,
University of Cape Town, Centre for Occupational and
Environmental Health Research, Cape Town, South
Africa; 2National Institute for Occupational Health,
Immunology, Johannesburg, South Africa; 3Molecular
Immunology, Comparative Genomics Centre, Faculty of
Medicine, Health and Molecular Sciences, James Cook
University Townsville, Queensland, Australia
Background: Previous epidemiological stud-
ies have reported obstructive lung disease
due to inhalation of spices. The aim of this
study was to determine the prevalence of
occupational allergic respiratory disease and
associated risk factors in spice mill workers.
Method: Exposure metrics were developed
from 62 full shift personal samples analy-
sed for inhalable particulate mass, spice
dust allergen (garlic) and endotoxin using
ELISA inhibition and chromogenic LAL
assays. A cross-sectional study of 150
workers used ECRHS questionnaires,
Phadiatop, specic IgE to occupational
allergens (garlic, chilli pepper, wheat) (Pha-
dia, ImmunoCAP), spirometry and FeNO
as per ATS/ERS criteria (2005).
Result: The mean dust particulate (GM
2.06 mg/m3, LOD-47.64) and garlic allergen
(GM 0.24 lg/m3, 0.0243.29) levels were
higher in blending, while endotoxin (GM 60
EU/m3, 23390) elevated in milling areas.
Dust particulate correlated more strongly
with garlic allergen (r = 0.70) than endo-
toxin (r = 0.43). The mean age of workers
was 33 years, 71% were male, 46% current
smokers and 45% atopic. Spice dust related
ocular-nasal symptoms (43%) were more
common than asthma symptoms (17%).
Sensitisation to garlic (19%) was higher
than to wheat (9%) or chilli pepper (6%),
but highly correlated (r = 0.890.96).
COPD (FEV1/FVC <70%) prevalence was
6%, while 4% had airow reversibility
(FEV1 >12%) and 8% airway inamma-
tion (FeNO >50 ppb). Allergic rhinitis due
to garlic (9%) was higher than to wheat
(5%) or chilli pepper (2%). Probable
asthma (FeNO >50 ppb) due to garlic
(4%) was higher than to wheat or chili pep-
per (3%). In multivariate models, work-
related ocular-nasal symptoms due to spice
dust (OR = 2.3) were associated with
airborne garlic allergen (>0.235 versus
<0.066 lg/m3) and work-related asthma
symptoms with garlic allergen (OR = 3.2)
and endotoxin (OR = 5.2) (>59.06 versus
<44.86 EU/m3). Spice dust related asthma
symptoms were also associated with garlic
sensitisation (OR = 4.7). Probable asthma
was more strongly associated with chili pep-
per (OR = 23.9) sensitisation than wheat
(OR = 6.4) or garlic (OR = 5) as was
COPD in relation to chilli pepper (OR =
15.6) compared to wheat (OR = 7.5) or
garlic (OR = 4.9) sensitisation.
Conclusion: Workers exposed to inhalable
spice dust particulate (mean >2 mg/m3)
containing allergens as low as 20 ng/m3
have an increased risk of work-related
lower respiratory symptoms, probable
asthma and COPD. The strongest associa-
tion is with chilli pepper sensitisation.
1578
Cleaners asthma - does it really exist?
Lipinska, A1; Wittczak, T1; Wiszniewska, M1; Palczynski,
C1; Nieradka, A2; Walusiak-Skorupa, J1
1
Nofer Institute of Occupational Medicine, Department
of Occupational Diseases, Lodz, Poland; 2Poland
Background: Asthma is the most common
occupational allergic respiratory disease.
The results of several current researches
reveal also high risk of work-related
asthma among cleaners. The aim of study
was to compare the frequency and clinical
symptoms due to hypersensitivity to clean-
ing agents at the workplace.
Method: Study groups comprised: 103
cleaning workers randomly examined in
their workplace (group 1) and 20 patients
of our Department - cleaning workers with
suspicion of occupational asthma (group
2). Questionnaire, skin prick tests (SPT) to
common allergens and chemicals, total and
specic serum IgE (asIgE) and rest spirom-
etry were performed in all subjects. Specic
provocation test were performed in cases
of suspicion of occupational asthma.
Result: 67% subjects from group 1
reported symptoms of allergic disease,
mainly rhinitis (29.1%), dyspnoea (26.2%),
conjunctivitis (22.3%), skin symptoms
(21.3%) and chronic cough (14.6%). 58%
symptomatic subjects developed symptoms
during cleaning work. 19 (18.5%) subjects
had positive SPT to common allergens,
mostly to D. farinae (11.7%) and D. pter-
onyssinus (9.7%). Serum specic IgE to
disinfectants was not found. FEV1%FVC
less than 70% was found in 10 subjects
(9.7%). In group 2 all subjects reported
symptoms of allergic disease, mainly cough
(75%), dyspnoea (70%), rhinitis (50%) and
skin symptoms (50%), conjunctivitis
(40%). 8 (40%) subjects had positive SPT
to common allergens, mostly to D. pter-
onyssinus (25%) and D. farinae (20%).
Positive SPT to at least one occupational
allergen were found in 6 subjects (30%). In
1 person (5%) serum specic IgE to disin-
fectants mixture was present. Occupational
asthma was recognised only in one patient
in group 2.
595
 Poster Session 67 Occupational allergy

Conclusion: Most of respiratory symptoms effects of chemicals used during cleaning

reported by cleaners are not conrmed by activities seem to be the main problems in
objective diagnostic tests. Hypersensitivity that group.
to mites and increased risk of irritating

596 Allergy 66, Suppl. 94 (2011): 482642  2011 John Wiley & Sons A/S

1579
Allergic reaction to montelukast
Sanchez Salguero, C1; Chacon Parejo, A2; Molina
Gonzalez, A2; Cano Hoyos, M2
1
Puerto Real University Hospital, Paediatric Allergy
Department, Puerto Real, Spain; 2Spain
Introduction: One drug that has revolution-
ized the treatment of asthma in recent
years is Montelukast. Its mechanism of
action is based on blocking the CysLT 1
receptor preventing the action of cysteinyl
leukotrienes (LTC4, LTD4, LTE4) released
by mast cells and eosinophils and cause
bronchoconstriction, mucus secretion, vas-
cular permeability and accumulation of eo-
sinophils.
Case report: A man with 5 years of age
who had a diagnosis of asthma partially
controlled with inhaled budesonide at
doses of 800 mcgr/day, and also needed to
use inhaled salbutamol 1 per month for 2
3 days. After consulting the parents jointly
decided to start using Montelukast at
doses of 5 mg/day orally. In his personal
history had breastfeeding until the sixth
month of life with allergies to cows milk
the rst and second year. His mother had
been treated with Montelukast and utica-
sone + LABA inhalers during pregnancy
for asthma not controlled. When adminis-
tered the fth dose of Montelukast admit-
ted to the emergency at the hospital with a
picture of widespread distribution urticar-
ial lesions, swelling of lips, cough, and
malaise. Upon arrival is channeled via
peripheral venous adrenaline is given sc, iv
steroids and iv dexchlorpheniramine,
improving the patients general condition
gradually. No oxigenoterpia needed. In
additional studies performed urgent blood
count showed eosinophilia of 11%, normal
coagulation, biochemistry normal GOT
and GPT 67 45. Allergy Consultation sub-
sequently determined IgE 1340 IU/ml with
negative CAP to foods. I studied environ-
mental allergens by prick positive to mites.
Was performed PRICK-PRICK with a
preparation of Montelukast as a transport
using sterile vaseline. Montelukast
presented positive reaction with wheal of
0.8 0.5 mm (positive control of 0.5
0.5 mm). Then we proceeded to conduct
controlled exposure test and applying
the drug Montelukast 5 mg+Vaseline on
the lips developed edema of the lips with-
Allergy 66, Suppl. 94 (2011): 482642  2011 John Wiley & Sons A/S
Poster Session 68
Drug induced anaphylaxis
out respiratory distress and suspended the
test.
Conclusions: Montelukast in a drug used
in the treatment of asthma with inhaled
corticosteroids or individual therapy. It
also allows lower doses of inhaled corticos-
teroids to avoid side effects. But sometimes
presents adverse reactions, including ana-
phylaxis present in fewer than 1% of
patients treated. In these cases you have to
suspend the administration of Monteluk-
ast.
1580
Selective IgE-mediated hypersensitivity
to the methylprednisolone
Callero Viera, A1; Martnez Tadeo, J2; Perez Rodriguez,
E2; Hernandez Santana, G2; Rodriguez Plata, E2; Garcia
Robaina, J2
1
Nuestra Senora de Candelaria University Hospital,
Allergy Departament, Santa Cruz, Spain; 2Nuestra
Senora de Candelaria University Hospital, Allergy
Departament, Santa Cruz, Spain
Background: There are no many publica-
tions about IgE-mediated hypersensitivity
to corticoids. Those found are mainly type
IV reactions and topical cases. Herein we
present a case report of IgE-mediated
hypersensitivity to methylprednisolone
without cross-reactions to other corticoids
by topical, intramuscular or intravenous
administration.
Methods: The patient, a 25-year-old
woman, has been twice under intramuscu-
lar treatment with methylprednisolone.
5 min after the second dose, she presented
cough, dyspnoea sensation, thoracic tight-
ness, pharynx itch and labial and upper
and lower eyelids edematization. She was
treated with adrenaline and with aerosol
and oxygen therapies improving during the
following 12 h until the complete resolu-
tion 24 h later. Then she needed corticoids
treatment with hydrocortisone e.v. and
inhaled budesonide without showing any
symptom after its use. We plan a titration
of methylprednisolone and diluent as well
as a skin test to other commercialized e.v.
or im. corticosteroids. A simple-blind pla-
cebo-controlled oral provocation test
(OCPT) to deazacort is also programmed.
Results: Negative allergy skin prick test to
methylprednisolone. Negative results in the
allergy skin prick and intradermal tests 1/
10 to triamcinolone, betamethasone, dexa-
methasone and hydrocortisone. Positive
results in the intradermal tests to methyl-
prednisolone 1/10000; 1/1000; 1/100 and 1/
10. OCPT to deazacort: tolerance to a
1 mg/kg dose.
Conclusions: Due to the tolerance to e.v.
hydrocortisone, inhaled budesonide and
deazacort by systemic treatment; we think
this case suggests exclusively type I hyper-
sensitivity to methylprednisolone, without
showing cross reaction to other corticoids.
1581
Metamizol and sulpiride anaphylaxis
Alvarado, M1; Camara-Hijon, C2
1
Ciudad de Coria Hospital, Allergy Department, Coria,
Spain; 2Caceres Hospital, Immunology Department,
Caceres, Spain
Background: We study a women that was
to the hospital because she had renal colic,
doctors administer her one metamizol and
one sulpiride ampoule and she presented
immediately intense palms pruritus, wide-
spread itching, rash, hives and edema, also
she was having difculty swallowing and
breathing. She tolerates ibuprofen.
Methods: Cutaneous tests with metamizol,
oral provocation with sulpiride and baso-
phil activation test and cross-inhibition
with serafosa were realized.
Results: Prick: Negative, 1, 10 mg intrader-
morreaction: Positive with metamizol. Oral
provocation with 17 mg of sulpiride was
positive: The patient experimented intense
itching in her hands and hives in arms,
back and waist. Basophil activation test
was positive with both drugs, the sulpiride
reactivity was better and percentage of
activated basophils after allergen stimula-
tion higher than metamizol. Cross-inhibi-
tion with both drugs stick to resin was
negative.
Conclusion: Our patient was sensitized to
both drugs. She has allergy to metamizol
and sulpiride because she has double sensi-
tization or cross-reaction.
597

1582
Anaphylaxis to pyrazinamide: skin test
positivity and successful desensitisation
Bavbek, S; Yilmaz, I; Aydin, O; Kepil Ozdemir, S
Ankara University, School of Medicine, Department of
Chest Diseases, Division of Immunology and Allergy,
Ankara, Turkey
Background: Here we report a case of
PZA-induced anaphylaxis. To our best
knowledge, this is the rst reported case of
PZA-induced anaphylaxis proved by skin
prick test and oral provocation/desensitiza-
tion. A 44-year-old woman with a recent
history of hypersensitivity reaction to antit-
uberculous drugs was admitted to our
department. She reported that she had
developed diffuse itching particularly on
palmar and plantar region, ush, shortness
of breath, fainting, loss of consciousness
within 5 min after rst dose of rifampicin
(RIF), isoniazid (INH), PZA and ethambu-
tol (EMB). She had been immediately
admitted to the emergency department.
Her systolic blood pressure had been mea-
sured as 50 mmHg and had been treated
with oxygen and some medications of
which she could not recall. All drugs had
been stopped and patients had been
referred to our department.
Methods: Skin prick tests were performed
with PZA tablet, RIF capsule, INH tablet
and EMB tablet by prick to prick method
along with positive control and negative
control. Considering the negativity on skin
test, graded oral challenge tests were per-
formed with RIF capsule, INH tablet, and
EMB tablet. PZA was given by desensitiza-
tion protocol.
Results: Positive reaction was only seen
with PZA with a wheal of 3 3 mm sur-
rounded by erythema of 5 mm after
15 min. Skin prick tests with PZA were
negative in 10 healthy controls. No reac-
tion developed during oral provocation
tests and were accepted negative for tested
drugs (INH, RIF, ETH). The rst day of
desensitization was uneventful but the
patient developed a generalized rash and
pruritus (especially on palmar and plantar
region), severe headache with 1500 mg
cumulative dose of PZA on second day.
Next day total dose of 1500 mg PZA, was
given by divided dose of 750 mg with 1 h
intervals under premedication with cetiri-
zine. The patient was informed of drug
allergies and the use of these drugs and has
started to take quadruple antituberculosis
treatment asymptomatically.
Conclusions: Anaphylaxis induced by PZA
is a possible hypersensitivity reaction to
this commonly prescribed antituberculous
drug. This case illustrates both the benets
of skin test in the diagnosis of PZA
induced anaphylaxis and administration of
oral desensitization when dealing with a
598
serious and potentially fatal pathology
such as tuberculosis, guaranteeing that the
patient had the optimum antituberculous
therapy.
1583
Anaphylaxis: A 10 year review of report-
ing to the Portuguese pharmacovigilance
authority
Ribeiro-Vaz, I1; Marques, J1; Gomes, E2
1
Faculty of Medicine of University of Porto, Northern
Pharmacovigilance Centre, Porto, Portugal; 2Porto
Hospitalar Centre, ImmunoAlergology Department,
Porto, Portugal
Background: Anaphylaxis is a potentially
fatal systemic Adverse Drug Reaction
(ADR). It is classied as a type-B reaction
as it is an unpredictable and mostly dose-
independent event that occurs suddenly
after contact with the causative drug. Our
objective was to characterize a case series
of anaphylactic reactions reported to the
Portuguese Pharmacovigilance authority
during the past 10 years. Demographic
data of the patients and the implicated
drugs were analyzed as well as severity and
time trends.
Methods: Retrospective analysis of every
episode of anaphylaxis, dened according
the second symposium on the denition
and management of anaphylaxis criteria,
reported to the Portuguese Pharmacovigi-
lance System between 01.01.2000 and
01.11.2010. Report sources are health-care
professionals and marketing authorization
holders.
Results: We found 918 cases of anaphy-
laxis that met the proposed criteria. Age of
the patients at the episode varied from
7 days to 91 years (mean: 48 years, SD:
21 years) with 87 cases (9%) under
18 years. There is an overall female pre-
dominance (67% women) but male were
predominant in pediatric ages (56%).
There was a trend in increasing reporting
and 31% (284) of the cases were reported
during the last 2 years (20092010). 19%
of the episodes led to hospitalization and
we found 24 (3%) cases with a fatal out-
come (four antibiotics, four antineoplastic
drugs, three NSAIDs). Antibiotics were the
group responsible for most of the reported
reactions (17%) followed by NSAIDs/
Acetaminophen (13%). Antineoplastic/cito-
toxic drugs occupied the 3rd place and
were followed by immune-modulators, this
group with an important increase in the
last 3 years. Vaccines and radiographic
contrast media were also important con-
tributors with 60 and 40 reports respec-
tively.
Conclusion: In this series of drug related
anaphylaxis we found as other authors that
most of the reported episodes were associ-
Allergy 66, Suppl. 94 (2011): 482642  2011 John Wiley & Sons A/S
Poster Session 68 Drug induced anaphylaxis
ated with widely used drugs (sometimes
over the counter) as antibiotics and analge-
sics. Anaphylaxis can occur at any age.
The female gender was more represented
except for pediatric patients as in other
previous studies. The important proportion
of serious cases is probably due to selec-
tion bias.
1584
Anaphylaxis to histamine H2-receptor
antagonist
Moreno-Fernandez, A1; Figueroa, C2; Ruiz-Hornillos, F3;
Henriquez, A3; Beitia, J4; Mira, J1
1
Virgen de la Luz Hospital, Allergy, Cuenca, Spain;
2
Colmenar SAR, Emergency, Spain; 3Infanta Elena
Hospital, Valdemoro, Spain; 4Guadalajara Hospital,
Spain
Background: Histamine H2-receptor antag-
onists reduce acid gastric secretion. They
are used in de treatment of peptic ulcer
disease and gastroesophageal reux dis-
ease. Nizatidine is one of these drugs. It is
widely used; however hypersensitivity reac-
tions are rare.
Methods: We report on a 62-year-old man
who after administration of one tablet of
nizatidine and one tablet of diclofenac suf-
fered palm, sole and genital pruritus.
Afterwards, it extended itself together with
urticaria affecting the whole body, labial
angioedema along with, giddiness and
unconsciousness. The patient had to be
admitted for 24 h to be treated with corti-
costeroids. Informed consent was obtained
from the patient. Skin prick-tests with dic-
lofenac (2.5 mg/ml) and nizatidine (0.3 mg/
ml) and intradermoreaction with these
drugs at the concentration of 0.25 and
0.03 mg/ml respectively were realized Sim-
ple-blind controlled oral challenge were
realized with increasing doses of aspirin
and nizatidine.
Results: All cutaneous tests were negative.
Simple-blind controlled oral challenge with
aspirin up to therapeutic dose resulted neg-
ative. Because of the negativity of the cuta-
neous tests, we decided to perform a
controlled oral challenge with 75 mg of
nizatidine. After 15 min, the patient suf-
fered a very intense palm, sole and genital
pruritus along with conjunctival injection
and generalized erythema. All the symp-
toms went on increasing despite the treat-
ment with epinephrine and corticosteroids.
The patient had to be monitored in emer-
gency unit where it was detected hypoten-
sion.
Conclusions: We report on a case of ana-
phylaxis due to nizatidine shown after a
controlled oral challenge with a small dose
of this drug despite negative cutaneous
tests.
Poster Session 68 Drug induced anaphylaxis
1585
Management and evaluation of specific
anaphylactic and anaphylactoid reactions
in teenagers
Bajraktarevic, A1; Mahinic, A1; Maglajlija, S1; Begovic,
B2; Dzinovic, A3; Begic, Z4; Niksic, H5; Fako, S5; Djulepa
Djurdjevic, A6; Resic Secerbegovic, A7; Hadzihasanovic,
E8
1
Public Health Institution of Canton Sarajevo, Pediatrics
Department, Sarajevo, Bosnia and Herzegovina;
2
Clinical Medical Center Sarajevo, Department for
Clinical Pharmacology, Sarajevo, Bosnia and
Herzegovina; 3Pediatrics Clinic Sarajevo, Department
for Allergology and Pulmonology, Sarajevo, Bosnia and
Herzegovina; 4Pediatrics Clinic Sarajevo, Cardiology
Department, Sarajevo, Bosnia and Herzegovina;
5
Pharmacy Faculty Sarajevo, Clinical Pharmacology,
Sarajevo, Bosnia and Herzegovina; 6General Hospital
Sarajevo, Emergency Department, Sarajevo, Bosnia and
Herzegovina; 7Dermatologic Clinic Sarajevo,
Allergology Department, Sarajevo, Bosnia and
Herzegovina; 8First Medical Aid of Canton Sarajevo,
Pediatrics Department, Sarajevo, Bosnia and
Herzegovina
Background: Although two reactions, ana-
phylactic and anaphylactoid reactions, dif-
fer in their underlying pathophysiology the
clinical manifestations and treatment are
the same in both cases. The management of
anaphylactic and anaphylactoid reactions
must be prompt. The diagnostic approach
to the teen agers patient should parallel
those used in diagnosis of other adverse
reactions to allergens and anaphylaxis.
Objective of this study to determine the
current frequency of accidental anaphylac-
tic and anaphylactoid reactions and expo-
sures occurring in allergic children and
identify factors associated with exposure.
Methods: Statistics calculation of data for
teen agers children population during rst
decade of millennium we made by Sigmas-
tat-3 for about 100 cases. Medical charts
of teen agers children evaluated and diag-
nosed as having anaphylactic allergy reac-
tions in pediatrics primary, secondary,
tertiary care and allergy and immunology
clinic in Canton Sarajevo.
Results: The severity of reaction was be
within minutes of exposure to triggering
substance, or delayed for up to half an
hour. Classical symptoms included diffuse
erythema (20%), pruritus (90%), urticaria
(45%), angio-oedema (40%), broncho-
spasm (75%), laryngeal oedema (50%),
vomiting (33%), hypotension (80%) and
cardiac arrhythmias (45%) in our study
during last ten years 20002010 for ana-
phylactic reaction in teen agers population.
Discussion: A meta-analysis of these studies
concluded that a drugs and food allergen
avoidance diet of mothers during lactation
may substantially reduce the development
of allergy and anaphylactic reactions in
their child in teen agers childhood.
Conclusion: The key of successful treat-
ment of anaphylactic and anaphylactoid
reaction is adrenalin. The exact cause of
someone developing an anaphylactic
Allergy 66, Suppl. 94 (2011): 482642  2011 John Wiley & Sons A/S
allergy reaction is unknown. Being allergic Conclusions: We report a case of systemic
to allergen from food, drugs, chemical sub- reactions to macrogol 4000 and 6000
stances or something from environment is occurring in the same patient after the
fairly common especially in children. intake of different drugs. Even if the
Keywords: Anaphylaxis, Allergy, Children, monomeric ethylene oxide specic IgE
Management were negative, the positive skin prick test
to macrogol 4000 and to paracetamol
syrup (macrogol 6000) suggested a type 1
reaction. We suspect that allergic reactions
1586
to macrogols may be underreported.
Systemic reactions to polyethylene glycol
4000 and 6000: a case report
Bommarito, L; Mietta, S; Nebiolo, F; Rolla, G
A.O. Mauriziano Umberto I, Allergy and Clinical 1587
Immunology department, Turin, Italy
Anaphylactic reaction to ceftriaxone dur-
ing general anesthesia
Background: Polyethylene glycol (macro-
Djuric, V; Raskovic, S; Peric Popadic, A; Tomic Spiric, V;
gol), is a polymer of ethylene oxide and Sojic, J; Durutovic, D; Delic, R; Plavsic, A; Spasic, S;
water which is widely used in food, cos- Bogic, M
metics and drugs because of its stabilizing Clinical centre of Serbia, Clinic for allergology and
immunology, Belgrade, Serbia
property. Hypersensitivity reactions have
been reported after parenteral and oral
Background: Anaphylactic reactions during
administration of products containing mac-
general anaesthesia may be due to various
rogol (corticosteroid, electrolyte lavage and
drugs and other substances, including anti-
evacuant solutions, tablets).
biotics. Epidemiological studies from
Case report: A 46-year-old man without
France show that neuromuscular blocking
previous history of allergy referred to our
agents are the most important cause but
outpatient clinic for suspected drug reac-
antibiotics, particularly beta lactams, are
tions. He reported mild urticaria and mouth
now recognized as the third most frequent
itching in 2007 a few minutes after the
cause of anaphylactic reaction.
intake of nimesulide granules (containing
Methods: We present a case of a 36-year-
macrogol 4000); a few months later, follow-
old woman in whom orthopedic operation
ing paracetamol syrup one dose (containing
of her scoliosis was planned. Midazolam,
macrogol 6000), he rapidly developed gener-
propofol, ramifentanyl, atropine, and rocu-
alized urticaria, face and neck angioedema
ronium were administred. Several minutes
and dyspnoea, requiring medical assistance
after ceftriaxone was injected intrave-
in ER. Metamizol and ibuprofen (without
nously, arterial hypotension and skin
macrogol) were well tolerated. He also
erythema developed and operation was
reported erythema using some types of bath
cancelled. She was referred to allergologist.
soap and shaving cream.
Results: She had no history of drug
Method and Results: Standard (ALK-
allergy. She also had no allergic reaction
Abello), shaving cream series (cetylic alco-
when she was in general anaesthesia twice
hol, stearylic alcohol, cinnamic alcohol,
before. Skin tests with atropine, propofol,
camphor, menthol, sorbitan monolaurate,
midazolam, ramifetanyl, rocuronium were
triethanolamine) and polyethylene glycol
negative. Test with latex glove was nega-
patch tests were performed, resulting posi-
tive. Skin prick tests with common aeroal-
tive (++) thimerosal, balsam of Peru and
lergens were negative. Intradermal test
menthol. Prick test with paracetamol syrup
with ceftriaxone was positive.
gave a positive reaction with a wheal
Conclusion: We conclude that anaphylactic
increasing up to 30 mm diameter after
reaction during general anaesthesia was
30 min, when face and neck erythema
caused by ceftriaxone.
developed with mild throat closure sensa-
tion. One month later, skin prick test with
injectable paracetamol (starting from 1/
10000 to 1/1) were performed with negative 1588
results. The graduated oral challenge with Idiopathic anaphylaxis
paracetamol tablets was tolerated up to the Geraldo Dias, J1; Sousa, F2; Luz, S1; Prates, S3; Leiria
1 g cumulative dose. Specic ethylene oxide Pinto, P3
IgE (CAP-Phadia) was negative. Skin prick
1
Hospital Santa Maria, Centro Hospitalar Lisboa Norte,
Immunoallergology Department, Lisbon, Portugal;
test with macrogol 400 (100 mg/ml from 1/ 2
Hospital Central do Funchal, Immunoallergology
10000 to 1/1) was negative, while skin prick Department, Funchal, Portugal; 3Hospital Dona
test with macrogol 4000 (100 mg/ml) at 1/ Estefania, Centro Hospitalar Lisboa Central,
Immunoallergology Department, Lisbon, Portugal
10000 dilution was positive (wheal diameter
6 mm). Skin prick test with paracetamol
Background: Anaphylaxis is a severe aller-
syrup, macrogol 400 and 4000 tested nega-
gic reaction that is rapid in onset and may
tive in 10 non atopic controls.
cause death. It is essential to identify its
599

etiology, because its treatment involves
triggers avoidance. Idiopathic anaphylaxis
is a diagnosis of exclusion. It is necessary
to rule out possible association with food,
drugs, radiocontrast media, insect venoms,
latex or exercise.
Case report: Eight-year-old boy with per-
sonal history of asthma and persistent non-
allergic rhinitis taking montelukast,
levocetirizine and nasal budesonide daily.
Skin prick tests (SPT) with aeroallergens:
negative; lung function testing: normal. At
6-year-old, experienced his rst episode of
anaphylaxis (generalized urticaria, dyspnea
and vomiting), without any triggering fac-
tor identied. In the next 10 months, had
ve episodes of anaphylaxis, all of them
with unknown etiology and treated with
epinephrine at the hospital. His mother
denied any possible association with food
or drug intake. After an exhaustive ques-
tionnaire, she said that episodes were pre-
ceded by headache and usually medicated
with paracetamol or ibuprofen. She denied
any episodes of anaphylaxis associated
with ingestion of these drugs, stating that
there was no reaction with these drugs in
other situations. Laboratory: mild eosino-
philia; serum tryptase-3.6 ng/ml (normal).
We performed an oral provocation test
with paracetamol and 10 min after the
third dose (cumulative dose: 290 mg), he
developed trunk and limbs urticaria, fol-
lowed by cough and wheezing. It was
administered i.m. epinephrine, i.v. antihis-
tamine and corticosteroid. After 45 min,
the patient developed face angioedema,
urticaria got worse and i.m. epinephrine
was administered again, with symptoms
resolution in 3 h. He did not take paracet-
amol again and had no further episodes of
anaphylaxis. After the provocation test his
mother informed us that she had recently
administered him ibuprofen twice at home,
with no reaction.
Discussion: This case illustrates anamnesis
relevance in determining the etiology of an
episode of anaphylaxis. Drug etiology of
anaphylactic reactions was conrmed;
despite drug ingestion has been denied sev-
eral times. The patient should always carry
self-injectable epinephrine, because there
are a lot of over-the-counter drugs with
paracetamol in its composition, which
could be responsible for a new accidental
ingestion that may be fatal.
600
1589
Anaphylaxis with polyethylene glycol
conjugated asparaginase in two children
with acute lymphoblastic leukemia
Sahiner, U1; Yavuz, S1; Gokce, M2; Buyuktiryaki, B1;
Yilmaz, E1; Altan, I2; Aytac, S2; Tuncer, M2; Sackesen,
C1
1
Hacettepe University Faculty of Medicine, Pediatric
Allergy and Asthma Department, Ankara, Turkey;
2 for anaphylaxis.
Hacettepe University Faculty of Medicine, Pediatric
Hematology Department, Ankara, Turkey
Background: The chemotherapy agent L-
asparaginase is an important part of acute
lymphoblastic leukemia (ALL) treatment
despite the high rates of adverse drug reac-
tions. Polyethylene glycol conjugated as-
paraginase (PEG-ASPA) was developed to
minimize the adverse drug effects including
hypersensitivity reactions. In the literature
a very few cases with PEG-ASPA induced
anaphylaxis were reported. Premedication
and desensitization protocols may prevent
the development of anaphylaxis in almost
all cases. Herein we present two cases of
anaphylaxis with PEG-ASPA where one
developed anaphylaxis despite of the appli-
cation of both premedication and desensiti-
zation.
Cases: First patient was a girl aged 8-
years-old and diagnosed as CALLA (+)
B-cell ALL with CNS relapse. As she expe-
rienced anaphylaxis with native Escherichia
coli asparaginase we preferred to use PEG-
ASPA, according to the ALL REZ BFM
Protocole 2002. Premedication with meth-
ylprednisolone, hydroxyzine and ranitidine
was done followed by administration of a
2 h PEG-ASPA infusion protocol that was
1 IU administered intravenously and was
doubled every 10 min until the desired
total dose was reached. Because of the
pharmacologic properties of the drug we
could not prolong the desensitization pro-
tocol time beyond 2 h. Forty minutes after
the initial dose of infusion, she had pruri-
tus, generalized urticaria, dyspnea and
hypotension. Treatment of epinephrine and
bolus uids and corticosteroids rapidly
reversed the condition of the patient. The
second patient was a two and a half-year-
old boy diagnosed as CALLA (+) B cell
ALL. Since previously, he had grade two
allergic reactions to native l-asparaginase,
the therapy switched to PEG-ASPA.
According to the Modied St Jude T15
Protocole, he was administered PEG-
ASPA intramuscularly. Three hours after
the injection of PEG-ASPA he developed
generalized urticaria, dyspnea, vomiting
and hypotension (70/50 mmHg) he was
given antihistamines and corticosteroids
and followed for 24 h.
Conclusions: The primary case is the rst
case, who developed anaphylaxis with
PEG-ASPA despite of the prophylaxis with
Allergy 66, Suppl. 94 (2011): 482642  2011 John Wiley & Sons A/S
Poster Session 68 Drug induced anaphylaxis
premedication and desensitization proto-
cols. Pharmacologic properties of the drug
are a major limitation to develop more
effective desensitization protocols. The
anaphylactic reactions with PEG-ASPA
observed in these cases warn us to observe
more carefully the patients with PEG-
ASPA treatment in a clinical setting ready
1590
Anaphylaxis to non-steroidal anti-inflam-
matory drugs
Gonzalez-Martin, I; Botello-Borrego, M; Cabeza-
Rodriguez, N; Segura-Sanchez, C; Guardia-Martinez, P
Virgen Macarena University Hospital, Allergy
Department, Seville, Spain
Background: Propionic-acid derivative non-
steroidal anti-inammatory drugs (NSAID)
are extensively used as analgesic, antipy-
retic and anti-inammatory treatments.
Consequently, increased of adverse reac-
tions have been observed. In the present
report, we show the case of a woman who
developed anaphylaxis after Ibuprofen was
taken.
Methods: Case report:
A 50-year-old woman presented with
severe anaphylaxis 3 min after ibuprofen
was taken for a headache. Symptoms were:
rash, pharyngeal node, generalized exan-
thema, tongue angioedema, dyspnea and
unconsciousness. The patient was fully
recovered after 30 min of intravenous
treatment of Diphenhydramine, Epineph-
rine and Metilprednisolone. Subsequently,
she had a second anaphylactic reaction as
a consequence of an accidental ingestion of
Ibuprofen.
Results: The following tests were per-
formed: -Prick test with a standard battery
of air-allergens: Negative. -Prick test with
Ibuprofen and Ketoprofen : Negative -
Intradermal test with Ketoprofen: Negative
-Drug provocation testing with: ASA,
Clonixin, Piroxicam, Diclofenac: Negative.
Conclusion: Selective reaction to a NSAID,
such as ibuprofen, has increased in the last
years as a consequence of their extensive
use. There are two types of anaphylactic
reactions with NSAID: 1.Asthma and urti-
caria. These reactions are related to both
the mechanism of action of NSAID and
the cross-reactivity to NSAIDs. NSAID
mechanism of action is the inhibition of
prostaglandin production by blocking the
cyclooxygenase enzymes known as COX-1
and COX-2, thus shunting arachidonic-
acid metabolism toward the 5-lipoxigenase
pathway, resulting in increased production
and release of cysteinyl leukotrienes.
2.Allergic type reactions, which are most
likely mediated by Ig E. This kind of reac-
tion can be life - threatening and they are
Poster Session 68 Drug induced anaphylaxis
limited to a single drug or a chemically
closely related one. In this case report our
patient developed the second type of ana-
phylactic reaction. She was diagnosed hav-
ing allergy to Ibuprofen and others
derivates of propionic acid, in spite of hav-
ing a negative prick test, and because she
was not allergic to other groups of
NSAID. In order to diagnose this allergy
selective a single NSAID, despite having a
negative prick test, we should do a provo-
cation test with other groups of NSAID.
1591
Apparently idiophatic anaphylaxis
1 1
Ortega-Camarero, M ; Pinero Saavedra, M ; Avila
Castellano, M2; Gonzalez-Quevedo Tejerina, T1;
Cimbollek, S3
1
Hospital Universitario Virgen del Roco, Allergy,
Sevilla, Spain; 2Hospital Virgen del Roco, Allergy,
Sevilla, Spain; 3Virgen del Roco, Allergy, Sevilla,
Spain.
Background: Idiopathic Anaphylaxis (IA)
is a well-described syndrome of anaphy-
laxis without any recognised external trig-
ger. These patients show same symptoms
as patients with other types of anaphylactic
reaction. Attacks can occur with variable
frequency, and the symptoms usually are
urticaria or angioedema with or without
bronchospasm and/or hypotension. IA is a
diagnosis of exclusion.
Methods: We present a case of a 53-year-
old man who has had several episodes of
anaphylaxis in the last 2 years without any
recognized trigger. Patient referred that he
was for long being treated of hypercholes-
terolemia with a statin and he denied any
other disease and did not explain his symp-
toms associated with any intake of food or
drugs. Otherwise he had only contact with
a drug while preparing and giving to his
daughter a cefuroxime in solution. We
made skin prick test and intradermoreac-
tion to betalactams with histamine and
negative control. Specic IgE to betalac-
tams and other laboratory tests as activa-
tion basophils test (TAB) were made to
different concentration of the drugs. We
also tested tryptase. Tryptase can be very
useful in differentiating anaphylaxis from
many other conditions that can masquer-
ade as anaphylaxis.
Results: The cutaneous test (prick and
intradermal) with betalactams were positive
after 15 min to 2 mg/ml concentration of
cefuroxime prick (papule = 4 cm). Specic
serum IgE was negative to betalactams,
and BAT was positive to cefuroxime and
negative to the rest of betalactams. After
being diagnosed, the patient avoided expo-
sition and he hadnt needed any emergency
assistance and he didnt present any slight
sign or symptom of anaphylaxis.
Allergy 66, Suppl. 94 (2011): 482642  2011 John Wiley & Sons A/S
Conclusions: The inadverted exposition to of her allergy to povidone-iodine this
a drug is a frequent reason of anaphylaxis patient was treated antiseptically with
that it has to be had in account in the hydrogen peroxide (H2O2).
study of every IA. Only after a thorough
history and physical examination should
the diagnosis been entertained. Skin prick
1593
testing, RAST and other lab tests as ba-
Anaphylaxis to etoricoxib: A case report
sophils activation test (BAT), are usually
required to rule out culprit allergens and Popescu, A1; Greblescu, R2
1
Pop de Basesti Medical Centre, Bucharest, Romania,
the numerous diseases that may masquer- Romania; 2Medas Clinic, Bucharest, Romania
ade as IA. A proper history is probably
the most important tool in the work-up, Background: Etoricoxib is a nonsteroidal
because it will guide subsequent testing. anti-inammatory drug (NSAID) that
inhibits the inducible form of cyclooxygen-
ase (COX), COX-2 with high selectivity.
1592 COX-2 inhibitors are widely used and are
Anaphylaxis to antiseptics. A case report generally well tolerated, with low incidence
of adverse reactions. Although immediate-
Gonzalez-Martin, I; Jurado-Palma, F; Cabeza-Rodriguez,
N; Botello-Borrego, M; Ramirez-Jimenez, A; Guardia-
type hypersensitivity reactions induced by
Martinez, P etoricoxib and other COX-2 inhibitors are
Virgen Macarena University Hospital, Allergy rare, anaphylactic reactions have been
Department, Seville, Spain
reported.
Case report: A 47-year-old woman, with
Background: A widely used topical antisep-
no personal or family history of atopic dis-
tic is composed by povidone-iodine (poly-
ease, was referred to our clinic 3 months
vinyl pyrrolidone- iodine). In the last two
after an episode of generalised urticaria,
centuries Iodine has been used as a com-
angioedema of the lips, tongue and uvula,
mon treatment for injuries, however, only
dyspnea with wheezing, hypotension, and
a few cases of anaphylaxis reaction, as a
syncope that occurred 7 h after oral
consequence of povidone-iodine treatment,
administration of piroxicam (20 mg) and
have been reported. In most of the cases a
10 min after oral administration of etoric-
contact dermatitis is described after the
oxib (60 mg). The patient had been taking
antiseptic application.
piroxicam and etoricoxib intermittently
Methods: Case report:
over the past 6 months. She had been pre-
A 49-year-old woman with a cut on her
scribed a wide variety of NSAIDs includ-
hand was treated with povidone-iodine and
ing etoricoxib and piroxicam but also
a tourniquet to stop bleeding. After 15 min
ibuprofen, ketoprofen, nimesulide, meloxi-
of treatment she developed oedema in one
cam, celecoxib over the past 7 years for
nger, which subsequently spread to the
low back pain secondary to L4-L5 spinal
entire hand followed by hoarseness and
disc herniation. She denied any adverse
dysphonia. The signs disappeared after
drug reaction prior to this episode. A few
corticoid and antihistamines intravenous
weeks after the anaphylactic episode the
treatment in 24 h.
patient self-administered piroxicam at
Results: The following tests were per-
home and experienced no adverse reaction.
formed: Routine blood test: normal Com-
Skin prick tests (SPT) to aeroallergens
plement screening analyses: normal The
were negative. Laboratory ndings were
levels of the Thyroid hormone: normal.
unremarkable, with the exception of an ele-
Skin prick test with latex: negative. Patch
vated titre of antibodies to thyroperoxi-
test with Povidone Iodine: negative Patch
dase. Basophil activation test with
test (TRUE test): positive Nickel Sulphate.
etoricoxib has not been performed yet. The
Skin Prick test with Povidone Iodine
patient refused to consent to oral challenge
0.01% (0, 1 mg/ml): positive.
to etoricoxib.
Conclusion: Although anaphylaxis due to
Discussion: We present a patient who expe-
povidone-iodine can appear after surgery
rienced anaphylaxis to etoricoxib after hav-
its incidence in the population has not
ing previously tolerated the medication, as
been well studied. To date, only few a
well as another selective COX-2 inhibitor
cases describing this process have been
(celecoxib). This reaction was considered
reported in the literature. Usually, povi-
probable according to the Naranjo proba-
done-iodine administration causes skin
bility scale. We excluded hypersensitivity
reactions like contact dermatitis which are
to piroxicam because the patient self-
considered non-systemic reactions because
administered it at home with no adverse
it is considered a topical antiseptic. But,
reaction. Even though cases of hypersensi-
when Povidone-Iodine contacts the blood
tivity to etoricoxib are rare, clinicians must
system (surgeries, labour, incisions) these
consider the possibility of such reactions.
can cause a systemic reaction like the ana-
phylaxis reported in this case. As a result
601

1594
Anaphylaxis during caesarean section
due to carboprost
Makatsori, M; Scadding, G; Durham, S
Royal Brompton Hospital, Allergy Department, London,
United Kingdom
Background: Carboprost tromethamine is a
synthetic analogue of prostaglandin F2-
alpha that is licensed for the treatment of
postpartum haemorrhage (PPH). It is
administered intra-muscularly (i.m). The
most frequently reported adverse effects
are nausea, vomiting, ushing, pyrexia and
myalgia. A few cases of bronchospasm,
even in those without asthma have also
been reported.
Methods: A 39-year-old non-asthmatic
woman required an emergency caesarean
section and delivered a healthy child. She
had received oxytocin, cefuroxime, ondan-
setron, diamorphine, gelofusin, and lido-
caine. She then required an i.m. injection
of carboprost 250 mcg. Almost immedi-
ately after receiving this injection, she
experienced a sudden tightening in the
throat and difculty breathing. She was
noted to have developed lip and facial
swelling, an associated facial rash and a
raised erythematous swelling at the injec-
tion site. She promptly received treatment
with chlorpheniramine and hydrocortisone
and the symptoms then progressively set-
tled. Serum tryptase levels taken at 1, 5
and 24 h time intervals were negative.
Results: We performed skin prick testing
to the following substances: oxytocin, ce-
furoxime, latex, gelofusin, lidocaine,
chlorhexidine, ondansetron and carboprost.
These were all negative. Intradermal testing
(IDT) to cefuroxime, lidocaine, chlorhexi-
dine, ondansetron and oxytocin were nega-
602
tive. IDT with 1:100 concentration of
carboprost (2 mcg/ml) produced mild ery-
thema. IDT with the 1:10 and 1:1 concen-
tration produced strongly positive
reactions that were dose-dependent. Undi-
luted and 1 in 10 dilutions of carboprost
produced only very minor erythema when
tested on controls, but no typical allergic
wheal, as seen in our patient.
Conclusions: We present a case of anaphy-
laxis to carboprost, conrmed on intra-der-
mal testing. Our patient had never
knowingly received carboprost previously;
therefore the sensitizing event remains
unclear. Nevertheless, obstetricians and
allergists should be aware of the potential
for carboprost to cause anaphylaxis.
1595
Drug hypersensitivity anaphylaxis associ-
ated with primary biliary cirrhosis
Ribeiro, F; Carrapatoso, I; Faria, E; Segorbe Lus, A
Coimbra University Hospital Centre,
Immunoallergology Department, Coimbra, Portugal
Background: Drug hypersensitivity may be
associated with other pathologies, namely
auto-immune diseases.
Clinical case: We present a case of a
female patient, 47-year-old, that reported
three episodes of facial angioedema, gener-
alized urticaria, dyspnea, wheezing and
dysphagia, immediately after ingesting mul-
tiple unrelated foods (grilled meat, smoked
salmon, vegetable salad, prawns, wine,
scrambled eggs, turnip greens and sh). In
all of these episodes she also had taken
oral ibuprofen. After the anaphylactic epi-
sodes she begun an eviction diet of sh
and seafood but continued to ingest all
Allergy 66, Suppl. 94 (2011): 482642  2011 John Wiley & Sons A/S
Poster Session 68 Drug induced anaphylaxis
other foods. She also mentioned a previous
episode of angioedema and urticaria
related to the intake of a NSAID 15 years
ago. Concomitantly, she presented chronic
fatigue, arthralgia, back pain and multino-
dular goiter.
In order to overall access the patient
condition we performed cell blood count,
biochemical tests, serum immunoglobulins
and complement. To exclude a food
allergy, we carried out prick tests to sh
and seafood extracts. As the patient com-
plained of several unspecic symptoms,
compromising daily quality of life, auto-
antibodies were also performed. Finally, to
nd an alternative anti-inammatory drug,
patient underwent an oral challenge with
celecoxib. Immunoglubulins M and E and
c-GT were slight elevated (2.69 g/l,
283 UI/ml and 42 UI/l, respectively). Cell
blood count, complement and remaining
serum parameters were normal. Prick tests
to all allergens tested were negative.
ANCA auto-antibodies were positive,
namely anti-mithocondrials, thus leading
to the diagnostic suspicion of primary bili-
ary cirrhosis, later conrmed by liver
biopsy. Oral challenge with celecoxib was
negative.
Conclusion: When we have a patient pre-
senting symptoms suggesting a food or
drug hypersensitivity, we still have to
observe the patient as a whole in order not
to miss an important diagnosis such as an
auto-immune disorder. The association
between hepatobiliar dysfunction and the
use of NSAID drugs may have favored the
manifestation of hypersensitivity phenom-
ena to these drugs.

Desensitisation: an effective way of managing drug allergy
1596
Application of e-mail in successful
desensitisation to allopurinol
Bawakid, K1; Qutob, M2; Tayeb, M2; Koshak, E3
1
Ministery of Health, Allergy- Primary Health Care,
Jeddah, Saudi Arabia; 2King Abdulaziz University,
Family Medicine, Jeddah, Saudi Arabia; 3King
Abdulaziz University, Internal Medicine, Jeddah, Saudi
Arabia
Background: Oral desensitization in Allo-
purinol hypersensitivity can be helpful if
no alternative, but, it is associated with dif-
culties in monitoring the progress and to
initiate urgent response if needed. Our
objective to explore the efcacy of using
electronic mail (E-mail) to monitor oral
desensitization in a case with Allopurinol
hypersensitivity.
Methods: A 65-year-old Saudi gentleman
presented to the allergy clinic at King Ab-
dul-Aziz University Hospital as case of
Type I hypersensitivity reaction to Allopu-
rinol. Phase I desensitization was con-
ducted at the clinic using oral Allopurinol
suspension of 20 mg/ml starting from
0.01 mg up to 1 mg. Phase II desensitiza-
tion was conducted at home with increas-
ing doses from 1 mg up to 100 mg/day. A
daily patient follow up utilizing E-mail as
a communication tool was arranged and
mobile-phone calls were reserved for seri-
ous reaction.
Results: Both desensitization phases I and
II were successful over a period of 4 h and
5 weeks, respectively. He had minor allergy
symptoms (itching and rash) which
resolved completely with oral antihista-
mines and maintaining the same Allopuri-
nol dose until clear. Almost daily E-mail
messages were very helpful in the patient
monitoring and providing proper advices.
Conclusions: To our knowledge, this is the
rst case report of using E-mail as a tool
for secure follow up of successful oral
desensitization to Allopurinol. E-mail is
considered a modern and effective method
of communication between the patient and
the doctor, as it provides nearly a 24 h
medical support as long as no urgent
response is needed.
Allergy 66, Suppl. 94 (2011): 482642  2011 John Wiley & Sons A/S
Poster Session 69
available so the diagnose of the allergic
1597
reaction was done with the clinic manifes-
Desensitisation to cloroquine in a patient
tations of the patient, it was conrmed the
of systemic lupus erythematosus
st time that we tried the desensitization as
Daz Angulo, S; Alonso Castellano, M the patient presented again a delayed reac-
Centro Medico de Asturias, Allergy Department,
Oviedo, Spain
tion after 5 days of the total dose.
Table 1. For abstract 1597
Background: Cloroquine phosphate, a 4-
aminoquinoline compound, is an antima- Day Dose (mg)
larial and amebicidal drug. Hydroxychlo- 1 0.1
roquine sulphate is a synthetic antimalarial 2 0.2
drug that is widely used in rheumatologic 3 0.4
diseases, particularly important in the 4 0.8
treatment of systemic lupus erythematosus 5-11 1
12 2
(SLE). We preset a case of a patient with
13 4
SLE who presented a cutaneous pruritic
14 8
exanthematic reaction in her neck, back, 15-21 10
legs, arms and abdomen after 10 days of 22 20
treatment with hydroxycloroquine which 23 40
lasted 3 months although she stopped tak- 24-30 62.5 (1/4 of the tablet)
ing it and she was treated with corticoster- 31-34 187.2 (3/4 of the tablet)
oids and antihistamines. 35 250 (1 tablet)
Objectives: To make desensitization to
cloroquine in a patient with SLE who had
Table 2. For abstract 1597
presented a very long exanthematic reac-
tion and who needed the treatment. Day Dose (mg)
Methods: Skin prick test and patch test with 1 0.1
hydroxycloroquine were negative. We 2 0.2
decided to make a desensitization to cloroq- 3 0.4
uine as it was not possible to prepare hy- 4 0.8
droxycloroquine in solution and, according 5-11 1
to the North American Rheumatic Skin dis- 12 2
13 4
ease Study Group Organizing Committee,
14 8
cloroquine phosphate can be administered
15-21 10
to patients who have experienced prior hy- 22 20
droxychloroquine sulphate-associated exan- 23 40
thems with a low risk of the re-expression of 24-28 62.5 (1/4 of the tablet)
the exanthema or appearance of other clini- 29-39 125 (1/2 of the tablet)
cal forms. First desensitization: (Table 1) 40-50 187.5 (3/4 of the tablet)
After 4 days taking 250 mg the patient 51-60 250 (1 tablet)
presented again a general exanthematic reac-
tion and her general practitioner stopped
her medication. One month after we saw
her, we decided to make her a longer desen-
sitization. Second desensitization (Table 2) 1598
Results: The desensitization was done suc- Successful desensitisation with temozol-
cesfuffy and the patient didnt have any omide
reactions. Ruiz-Garcia, M; De las Heras, M; Hernandez, E; Sastre, J
Conclusion: The desensitization to cloroqu- Fundacion Jimenez Diaz, Allergy Department, Madrid,
Spain
ine in a patient of SLE who had presented
a delayed cutaneous reaction was done suc-
Background: Temozolomide is an oral al-
cessfully in 51 days. There are no refer-
kylating agent used for the treatment of
ences of skin test with cloroquine and
aggressive brain tumors such as grade IV
hydroxycloroquine, the threshold of the
astrocytoma, glioblastoma multiforme, and
specicity of patch test has to be deter-
melanoma.
mined and the specic serum IgE is not
603

Methods: We report a 57-year-old woman
with a history of gemistocytic astrocytoma.
She was started on temozolomide (Temo-
dal) 120 mg/day for 5 days each month
over six cycles. The rst cycle was tolerated
well. She presented 10 min after taking
temozolomide on the third day of the sec-
ond cycle pruritus, generalized erythema,
and angiodema resolved with systemic cor-
tecosteroids and antihistamines. Basal
serum tryptase and total IgE were 3.56 lg/
l and 89 UI/l, respectively. We performed
oral provocation challenge with 120 mg of
temozolomide presenting 60 min after the
same symptoms requiring intramuscular
adrenaline 0.3 mg, systemic corticosteroids
and antihistamines. No alternative treat-
ment for this patient existed, leading us to
perform a desensitization protocol with
temozolomide.
Results: On the rst-day of each cycle (ve
in total), the patient was admitted to our
hospital and was given cetirizine 10 mg, lo-
razepam 0.5 mg and ranitidine 150 mg 30
as premedication. Our dosage scheme was:
0.025 mg, 0.05 mg, 0.1 mg, 0.5 mg, 1 mg,
2 mg, 5 mg, 10 mg, 20 mg, 30 mg, 60 mg
and 110 mg at intervals of 30 min. The
maximum dose was set at 240 mg accord-
ing to her oncologist. Sixty minutes after
the last dose, she developed erythema and
pruritus in the palms, resolved in 20 min
with methylprednisolone 40 mg and dex-
chlorpheniramine 6 mg. The second day
she was given 120 mg and 120 mg at an
interval of 30 min; having tolerated this
well, she was discharged. At home, the
patient demonstrated tolerance to 240 mg/
day for the rest of the cycle.
Conclusions: To the best of our knowledge,
ours is the rst case of successful desensiti-
zation with temozolomide to be presented.
The patient was able to complete her che-
motherapy treatment successfully.
1599
Selective sensitisation to the highly
selective cyclooxygenase-2 inhibitor etor-
icoxib
Gonzalez Perez, R; Poza-Guedes, P; Matheu, V;
Sanchez-Machin, I; Iglesias, J
HUNS La Candelaria, Allergy Department, Santa Cruz
de Tenerife, Spain
Background: Specic sensitization to newly
developed selective cyclooxygenase-2
(COX-2) inhibitor drugs are infrequently
reported in the medical literature. A 63-
year-old lady was referred to our Institu-
tion presenting with a single episode of
skin adverse reaction described as acute
urticaria after the intake of etoricoxib.
Methods: Informed consent was obtained
and clinical evaluation conrmed that she
has been safely taken etoricoxib 90 mg for
604
Poster Session 69 Desensitisation: an effective way of managing drug allergy
more than 2 years. The onset after the
drug intake was 10 to 12 h, lasting 48 h
and showing full skin recovery after treat-
ment wi Guadalajara Hospital th i.v. ste-
roids and antihistamines. No systemic
symptoms were described. Allergy diagno-
sis work-up followed the ENDA Guide-
lines with a detailed clinical history, skin
tests, and single blind placebo controlled
provocations.
Results: A negative epi-cutaneous test
(patch test) with etoricoxib at 48 and 96 h
readings was followed by a skin prick test
(1/10) with etoricoxib showing a negative
immediate reading as well. A Single blind
placebo controlled challenge with increas-
ing doses of etoricoxib was positive at
60 mg cumulative dose. Skin urticaria was
well localised on her neck, chest and back,
the onset was 14 h and no anaphylactic/
systemic reactions were observed. Skin
biopsy conrmed the drug allergy diagno-
sis. Interestingly 3 months later a Single
blind placebo controlled challenge with cel-
ecoxib 200 mg was negative.
Conclusion: A non-immediate type cutane-
ous -urticaria- selective allergic reaction to
etoricoxib was conrmed. The lack of
cross-reactivity to another highly selective
COX-2 inhibitor celecoxib, in present clini-
cal case makes specic drug challenges of
utmost importance to achieve an accurate
diagnosis and provide -if possible- a safer
therapeutic alternative.
1600
Coexistence of allergy to human insulin
and protamine with successful specific
immunotherapy: a case report with
review of literature
Fazlollahi, M1; Hasani-Ranjbar, S2; Pourpak, Z1;
shokouhi, R1; Larijani, B2
1
Tehran University of Medical Sciences, Immunology,
Asthma and Allergy Research Institute, Tehran, Islamic
Republic of Iran; 2Tehran University of Medical
Sciences, Endocrinology & Metabolism Research
Center, Tehran, Islamic Republic of Iran
Insulin has an important role in the treat-
ment of diabetic patients. Further, it can
result in undesirable side effects. One of
the problems that are associated with insu-
lin therapy is allergic reactions. Insulin
allergy is a very rare complication of dia-
betes and management of this rare condi-
tion is very difcult. Here we report the
case of insulin and protamine sensitization
that was successfully desensitized using
rapid and long acting analogs. In addition
all case reports in literature were reviewed
and discussed. The patient was a 55-year-
old woman with poorly controlled type-2
diabetes and insulin allergy. She revealed
hypersensitivity reactions including urti-
caria and respiratory symptoms, immedi-
Allergy 66, Suppl. 94 (2011): 482642  2011 John Wiley & Sons A/S
ately after injection. The skin prick tests
(SPT) and intradermal tests (ID) revealed
sensitization to Protamine sulfate and all
insulins. So, specic immunotherapy with
glargine and aspart was done. Finally,
after specic immunotherapy we were able
to treat the patient with short- and long-
acting analogues successfully.
1601
Injection site reaction to etanercept: role
of skin test in the diagnosis of such
reaction and successful desensitisation
Bavbek, S1; Aydin, O1; Ataman, S2; Cahill, K3;
Castells, M4
1
Ankara University, School of Medicine, Department of
Pulmonary Disease, Division of Immunology and
Allergy, Ankara, Turkey; 2Ankara University, School of
Medicine, Department of PM&R, Division of
Rheumatology, Ankara, Turkey; 3Harvard Medical
School, Adverse Drug Reaction and Desensitization
Program, Allergy and Immunology Training Program
Brigham and Womens Hospital, Boston,
Massachusetts, United States; 4Harvard Medical
School, Adverse Drug Reaction and Desensitization
Program, Allergy and Immunology Training Program,
Brigham and Womens Hospital, Boston,
Massachusetts, United States
Treatment with etanercept is recommended
for patients with ankylosing spondylitis.
Injection-site reactions are the most com-
mon adverse reaction to etanercept; how-
ever the mechanism underlying these
reactions remains unclear. A 28-year-old
man with a history of reaction to etaner-
cept was admitted. He had been diagnosed
with ankylosing spondylitis and was
started on Humira (40 mg/0.8 ml, ada-
limumab) subcutaneous injections. He
developed local swelling, diffuse itching
and shortness of breath with Humira, and
the drug was discontinued. Then the treat-
ment was switched to Enbrel (25 mg eta-
nercept) 25 mg subcutaneously twice
weekly. He tolerated injections well for
3 months before he developed whole body
erythema with pruritus along with local
pruritus, redness and swelling 10 cm in
diameter at the site of injection within 10
15 min after the 22nd injection. Etanercept
was stopped and the patient was referred.
Prick testing with full strength of etaner-
cept (25 mg/1ml) was negative but intra-
dermal testing with 1/100 dilution of
etanercept resulted in a positive result with
a 7 7 mm wheal while histamine control
was 6 6 mm with negative result on late
reading of the intradermal test. Two
healthy controls were negative on intrader-
mal testing to this dilution. Because several
reports indicate that patients with injection
site reaction can be controlled with antihis-
tamines and do not need to stop the medi-
cation we decided to challenge the patient.
Therefore, we injected subcutaneously 1/10
Poster Session 69 Desensitisation: an effective way of managing drug allergy
of the total dose (2.5 mg) in the upper
thigh and the patient reacted with urticaria
over the whole trunk without local reac-
tion. The reaction was controlled with anti-
histamine within 1 h. Considering
positivity on skin testing and provocation
testing with etanercept, the drug was given
through a desensitization protocol. The
patient was pre-medicated each day with
aspirin 325 mg po, montelukast 10 mg po,
and famotidine 40 mg po 1 h prior to
desensitization and maintained on daily
Zyrtec. At present, the patient is on twice-
weekly, etanercept injection with Benadryl
25 mg po premedication and daily Zyrtec.
In our patient, positivity with etanercept
on intradermal testing and the patient
immediate reaction to the 1/10 diluted dose
of etanercept supported an immediate/pos-
sible IgE-mediated reaction. Therefore, we
desensitized the patient to etanercept. This
is, to our best knowledge, the rst reported
case of subcutaneous desensitization with
etanercept.
1602
Successful approach of breakthrough
reaction during desensitisation procedure
of a fixed drug eruption to allopurinol
Avila Castellano, R; Cimbollek, S; Ortega Camarero, M;
Pinero, M; Prados, M
Hospital Universitario Virgen del Rocio, U.G.C.
Intercentros Valme/Rocio de Alergologa, Sevilla, Spain
Background: Desensitization is a unique
approach for the management of xed
drug eruption (FDE). Only few cases have
been reported in the literature. Although
desensitization procedures breakthrough
reaction may occur there is no common
approach concerning management. We
describe a case of FDE to allopurinol suc-
cessfully desensitized despite breakthrough
reaction.
Methods: A 66-year-old man with history
of hyperuricemia and gouty arthritis previ-
ously diagnosed in our Unit of FDE to
allopurinol with negative skin prick test
and patch test was referred to our clinic
for allopurinol desensitization. Risks and
benets were discussed with patient and
informed consent was obtained. An outpa-
tient desensitization protocol was designed
according to previous regimen described.
50 mg of allopurinol was dissolved in
500 ml of distilled water with 14% sodium
bicarbonate. Starting dose of 10 mcg. and
doubling dose every 24 h until reaching
100 mg.
Results: At 25 mg patient started with
cutaneous lesions. Treatment was started
with Loratadine and protocol was modied
by reducing dose to previous step. After
3 days lesions started to clear. Doses were
Allergy 66, Suppl. 94 (2011): 482642  2011 John Wiley & Sons A/S
again doubled every 3 days until reaching
therapeutic dose of 100 mg. Skin lesions
disappeared completely progressively.
Conclusions: We report the rst break-
through reaction during desensitization
procedure of a FDE to our knowledge suc-
cessfully managed. Our case suggests that
decelerating the dose escalation and dupli-
cating doses at a slower rate may reduce
the incidence of reactions and improve tol-
erability.
1603
Efavirenz desensitisation in patients with
hypersensitivity to antiretroviral therapy
Testi, S1; Severino, M1; DErme, A2; Gola, M2; Iorno,
M3; Capretti, S3; Ermini, G3; Macchia, D3; Campi, P3
1
Azienda Sanitaria Firenze, Allergy Department,
Florence, Italy; 2Azienda Sanitaria di Firenze,
Dermatological Allergy, Florence, Italy; 3Azienda
Sanitaria Firenze, Allergy and Clinical Immunology
Unit, Florence, Italy
Background: The antiretroviral therapy has
improved the survival rate and quality of
life in patients infected with HIV; this ther-
apy is associated with adverse drug reac-
tions which include hypersensitivity
reaction, nephropathy, liver damage,
abnormal distribution of body fat and met-
abolic alterations. Hypersensitivity reac-
tions occur in 38% of HIV infected
treated patients and present an erythema-
tous, macupapular rash with or without
fever and sometimes severe systemic symp-
toms; rarely in some cases the reaction can
be fatal. These adverse reactions appear
during the rst 6 weeks of therapy: this
period may be shorter when the drug is
reintroduced. In patients without severe
systemic symptoms, an attempt in combi-
nation with antihistamines, should be made
to continue treatment. If this fails, desensi-
tization can be performed.
Methods: We describe two patients who
developed skin rash associated within the
rst 2 weeks after starting antiretroviral
therapy; the rst patient, male, 33 years
old, developed urticaria after atazanavir,
ritonavir, efavirenz; the second patient
(female 48 years old) experienced general-
ized pruritic rash after efavirenz, lamivudin
and zidovudin. We performed desensitiza-
tion for efavirenz (table 1) in 16-day proto-
col. During the rst 3 days the doses were
administred every 30 min in the morning;
then the dose were administred in the even-
ing, because its central nervous system dis-
turbance.
Results: The patients successfully com-
pleted efavirenz desensitization without
pretreatment with antihistamines or corti-
costeroids and they continue to have an
undectable viral load several months fol-
lowing desensitization.
Conclusion: In patients with a history of
hypersensitivity to multiple antiretroviral
drugs we can continue this therapy with
Efavirenz after appropriate desensitization.
1604
Development of a desensitisation proto-
col for a patient with Parkinsons disease
and with type 1 hypersensitivity to apo-
morphine
Foncubierta Fernandez, A1; Gutierrez Fernandez, D2;
Espinosa Rosso, R3; Anguita Carazo, J4; Fernandez
Melendez, S5; Leon Jimenez, A2
1
Joaquin Pece Health Center, Family Medicine, San
Fernando, Spain; 2Puerta del Mar University Hospital,
Neumology and Allergy, Cadiz, Spain; 3Puerta del Mar
University Hospital, Neurology, Cadiz, Spain;
4
Complejo Hospitalario de Jaen, Allergy, Jaen, Spain;
5
Carlos Haya University Hospital, Allergy, Malaga,
Spain
Background: Adverse skin reactions to
apomorphine have been described in less
than 1% of patients. We present the case
of a patient with Parkinsons Disease who
experienced episodes of generalised pruri-
tus and raised itchy wheals on underarms,
groin, chest and buttocks approximately
half an hour after the administration of
apomorphine. The symptoms resolved
within 20 min with antihistamine treat-
ment, leaving no residual lesions. As apo-
morphine was necessary as a rescue
therapy for the patient, we initiated a
desensitization regimen. To date, no other
reports of desensitization protocols to apo-
morphine have been described in the litera-
ture.
Methods: A Desensitization Protocol was
developed for the patient (Table 1). A
seven step regimen was established, with
increasing concentrations of apomorphine
at intervals of 15 min, up to the 10 mg/ml
concentration of the commercial prepara-
tion. The desensitization protocol was
implemented to the nal 3 mg dosage of
apomorphine previously demonstrated to
be the maximum required by the patient
for the improvement of his Parkinsonian
symptomatology. Premedication with
10 mg cetirizine was employed prior to
implementation of the protocol.
Results:
1. Total IgE<100 and specic
IgE<0.35.
2. Intradermal test with 10 mg/ml apo-
morphine: positive at 1:10 dilution.
3. Provocation test with 10 mg/ml apo-
morphine: positive after 20 min.
4. Provocation tests with sulphites of up
to 200 mg concentration: negative.
5. A single 3 mg subcutaneous injection
of apomorphine produced an
improvement in motor symptoms.
6. Desensitization Regime: A dose of
0.3 mg apomorphine produced slight
605

Table 1. For abstract 1604.
Step
(every Concentration Volume Total Cumulative Blood
15 min) (mg/ml) (ml) (mg) Total (mg) Pressure Rate Remarks
1 0.03 0.1 0.003
2 0.03 0.5 0.015
3 0.3 0.1 0.03
4 0.3 0.5 0.15
5 3 0.1 0.3
6 3 0.5 1.5
7 10 0.3 3 4.998
itching, which was treated with 5 mg
intravenous dexchlorpheniramine,
and did not require withdrawal of the
apomorphine; at step six the yawning
and dyskinesia experienced in the left
arm were attributable to side effects
of the apomorphine and not to an
allergic reaction. Having completed
the desensitization regimen, the
patient continued to receive continu-
ous daytime infusion for 12 h a day
at the rate of 1 mg/h and subse-
quently at 2.5 mg/h, with complete
tolerance to apomorphine.
Conclusions: We describe the rst desensiti-
zation protocol successfully applied to a
patient with type 1 hypersensitivity to apo-
morphine.
1605
Vitamin B 12 allergy and successful
desensitisation with cyanocobalamin:
a case report
Demirel, F; Kartal, O; Gulec, M; Yesillik, S; Caliskaner,
Z; Sener, O
Gulhane Military Medical School, Immunology and
Allergic Diseases, Ankara, Turkey
Background: Vitamin B12 deciency is
caused by either the inability of vitamin
B12 absorption, or lack of vitamin B12 in
the diet. The major clinical presentations
of vitamin B12 deciency are megaloblastic
anemia and gastrointestinal and neurologi-
cal symptoms. In the treatment of vitamin
B12 deciency cyanocobalamin and hy-
droxycobalamin are used which are syn-
thetically derived preparations of vitamin
B12. Allergy to vitamin B12 is rare, but
may be serious. We present a patient who
allergic to hydroxycobalamin and cyanoco-
balamin desensitized with cyanocobalamin
successfully.
Case: A 39-year-old woman with a history
of megaloblastic anemia due to deciency
of vitamin B12, applied with an intensely
pruritic generalized urticarial rash approxi-
mately 1 h after intramuscular injection of
1 mg cyanocobalamin. After 60 days from
606
Poster Session 69 Desensitisation: an effective way of managing drug allergy
1606
Successful rapid desensitisation to rabbit
Heart anti-thymocyte globulin in a girl with
aplastic anemia
0.003 100/62 91 Bobadilla-Gonzalez, P1; Zambonino-Carreiras, M1;
0.018 97/65 85 Perez-Rangel, I1; Garcia-Menaya, J1; Sanchez-Vega, S1;
0.048 98/65 89 Slight itching. Administration Cordobes-Duran, C2; Campano-Val, J3; Lopez-
Santamaria-Castro, C3; Corbacho-Hernandez, A3;
of 5 mg intravenous
Fuentes-Gutierrez, I3; Bajo-Gomez, R3
dexchlorpheniramine 1
Infanta Cristina University Hospital, Allergology,
0.198 112/71 80 Badajoz, Spain; 2University Hospital, Allergology,
0.498 110/70 78 Merida, Spain; 3Infanta Cristina University Hospital,
1.998 110/69 78 Yawning and dyskinesia in the Hematology, Badajoz, Spain
left arm
109/70 76 Background: Aplastic anemia is character-
ized by peripheral pancytopenia and bone
marrow hypoplasia. Election between
transplantation or immunosuppressive
the allergic reaction, skin prick tests were
therapy depends on availability of histo-
performed with cyanocobalamin and also
compatible family donor, patients age and
with hydroxycobalamin because of possi-
severity of aplasia. We present a case of
bility of cross reaction. Skin prick tests
aplastic anemia successfully desensitized to
were positive for both. Desensitization with
rabbit anti-thymocyte golublin (ATG).
the cyanocobalamin was planned as her
Methods: A fteen-year-old female was
B12 level was low and she required
diagnosed with aplastic anemia. Bone mar-
replacement therapy. Protocol of the
row transplantation was rejected as the his-
desensitization was performed according to
tocompatible brother suffered from mixed
literature(1) (Table 1). There were no
connective tissue disease. Treatment with
adverse reactions during the desensitization
rabbit ATG (for 5 consecutive days, at a
and a dramatic development was observed
rate of 25 mg/h by infusion pump of 6 h),
afterwards complete dose of vitamin B12
prednisone and cyclosporine was started.
therapy.
Premedication with paracetamol, methyl-
Conclusion: Vitamin B12 deciency is a
prednisolone and dexchlorpheniramine was
common disease and most of the patients
done. After 90 min, she presented pharyn-
require monthly intramuscular cyanocobal-
geal itching and dyspnea; the infusion was
amin or hydroxycobalamin injections for a
stopped, hydrocortisone was administered
long period. Although allergic reactions
and she improved within 30 min. Five
due to cyanocobalamin and hydroxycobal-
hours after the infusion was restarted dur-
amin injections are rare, there are a num-
ing 12 h, but after 90 min, she complained
ber of case reports describing this
of sore throat, breathlessness, chest tight-
situation. It is presumed that low molecu-
ness, wheezing and generalized hives, so
lar weight of B12 acts as a hapten for aller-
ATG was suspended.
gic reactions. In addition, the positive
Results: There was no history of allergies
results of skin test suggest an IgE mediated
or exposure to rabbit. Prick test with rab-
mechanism. Although there are a number
bit epithelium was negative. Specic IgE to
of case reports about allergic reactions to
epithelium and meat from rabbit were also
cyanocobalamin and hydroxycobalamin,
negative. Total IgE and tryptase were nor-
there is only one reporting desensitization
mal. Prick test with undiluted rabbit AGT
protocol and we conrm that case report.
(5 mg/ml) was negative, but intradermal
Desensitization protocol with cyanoco-
tests (1/105-1/100) were positive at 20 min.
balamin (1). 1.Caballero MR, Lukawska J,
Skin tests were not performed in controls
Lee TH, Dugue P. Allergy to vitamin B12:
for ethical reasons. Rapid intravenous
two cases of successful desensitization with
desensitization protocol described with
cyanocobalamin. Allergy. 2007 Nov;62(11):
other drugs was carried out with successful
1341-2.
result. The patient presented mild urticaria
the rst 3 days of treatment. Serum sick-
ness did not occur.
Conclusions: ATG is a polyclonal human
anti-thymocyte globulin used as immuno-
Table 1. For abstract 1605.
Day Dilution Amount(ml)
1 1/10000, 1/1000, 1/100, 1/10,1/1 0.1-0.3-0.6, 0.1-0.3-0.6, 0.1-0.3-0.6, 0.1-0.3-0.6, 0.1
7 1/1 0.01-0.03-0.1
21 1/1 0.1
49 1/1 0.1
Allergy 66, Suppl. 94 (2011): 482642  2011 John Wiley & Sons A/S
Poster Session 69 Desensitisation: an effective way of managing drug allergy

suppressive in organ and bone marrow
transplantation, and in aplastic anemia.
ATG can induce serum sickness and cyto-
kine release syndrome, but anaphylaxis is
infrequent. May be cytokine release could
justify the symptoms. Despite of positivity
of intradermal tests in immediate reading,
we can not state that it is due to an IgE
mechanism because controls are needed.
Few cases of desensitization to equine ATG
have been reported. To our knowledge, this
is the rst report of successful desensitiza-
tion to rabbit ATG. More studies are neces-
sary to clarify the involved hypersensitivity
desensitization protocol may be helpful for
patients with delayed hypersensitivity to
sorafanib.
1608
Tolerance induction using an ultra-rush
desensitisation protocol with glargine
insulin in a patient with an insulin allergy
and hypersensitivity to protamine
Costa, A; Morais Silva, P; Caiado, J; Barbosa, M
Santa Maria Hospital, Immunoallergology Department,
Lisbon, Portugal
changing insulin preparations fails to pre-
vent an allergic reaction, a rapid desensiti-
zation protocol may be a suitable
treatment option.
1609
Rapid rituximab desensitisation in a
patient with rheumatoid arthritis
Abadoglu, O1; Epozturk, K1; Atayik, E1; Kaptanoglu, E2
1
Cumhuriyet University, Faculty of Medicine,
Department of Immunology and Allergic Diseases,
Sivas, Turkey; 2Cumhuriyet University, Faculty of
Medicine, Department of Rheumatology, Sivas, Turkey

mechanism and the changes that occur dur- Background: Although rare, insulin allergy
ing the desensitization process. may occur in patients treated with subcuta- Background: Rheumatoid arthritis (RA) is
neous (SC) insulin preparations. In some an inammatory disease characterized by
cases, allergy may also develop to the addi- disturbances in T cell and B cell functions.
tives present in the preparation, namely Rituximab is a chimeric monoclonal anti-
1607 body (mAb) against CD20 that induces a
Desensititazion to sorafenib
protamine sulphate. When insulin is neces-
sary to the control of diabetes, desensitiza- profound depletion of B cells in the periph-
Olaverria Pujols, M; Iglesias Cadarso, A; Lopez San tion may be a treatment option. eral blood of RA patients. Desensitization
Martin, M; Reano Martos, M; Vasquez Bautista, A;
Case Report: A 45-year-old diabetic to rituximab has been described in case
Dionisio Elera, J
Hospital Puerta de Hierro, Allergy Service, Madrid, woman was referenced to our Immunoal- reports or small series of patients with he-
Spain lergology Department with complaints of matologic malignancies and some connec-
recalcitrant urticaria. She had started SC tive tissue diseases.
Background: Sorafenib is a small molecular human insulin 7 years earlier, reporting Methods: We report the case of a 32-year-
inhibitor of several tyrosine protein kinasas pruritic plaques at the injection site minutes old woman followed by our rheumatology
(VEGFR and PDGFR) and Raf, with after application and which persisted for department for RA for 7 years. In 2007,
antitumor activity in advanced hepatocellu- several days. Nine months before the refer- she had been given adalimumab in another
lar cancer and renal cell carcinoma. Sorafe- ral, her daily dose was increased and she medical center. After the second dose
nib is associated with a variety of adverse developed generalized urticaria. Concur- (15 days later from the rst dose), she had
cutaneous reactions like rash, hand-foot rently her blood sugar levels were increas- developed labial angioedema and tongue
skin reaction, and pruritus. Just a few ingly harder to control. Daily treatment swelling within 2 h of injection. Next
cases of hypersensitivity to sorafenib has with anti-histamines and changing insulin month, the medication was replaced by
been conrmed by positive skin tests. We preparations didnt control the symptoms. etanercept twice per week. After the fourth
report a successful oral desensitization in a Skin prick tests performed with several injection of etanercept the patient had pre-
patient with advanced hepatocellular carci- insulin preparations and protamine were all sented with tingling in her lips, syncope,
noma who developed a delayed hypersensi- positive. The same tests were negative when dizziness and headache and then she had
tivity to sorafenib. performed to a control group of ve dia- ceased etanercept therapy. Rituximab ther-
Methods: A male 60-year-old with hepato- betic insulin-treated patients with no his- apy was planned and skin tests for ritux-
cellular carcinoma presented pruritus and tory of reactions. The patient was treated imab were performed with a drop (10 mg/
maculo-papular rash in chest, arms and with SC insulin glargine using an ultra-rush ml) for prick test, and with 0.03 ml of
legs 2 weeks after taking Sorafenib. Eosin- protocol, under antihistamine therapy, 1:100 and 1:10 dilutions for intradermal
ophilia, renal or hepatic alterations were beginning with 0.1 U and a rapid augmen- test. All were negative. Rituximab was
not detected in peripherical blood analysis. tation in dose up to 6 U. Maintenance with given and 90 min into the infusion (175 mg
Cutaneous rash partially disappeared 6 U twice daily was well tolerated. of the expected 1000 mg dose), the patient
within 72 h after stop Sorafenib. A delayed Conclusions: When insulin allergy is estab- complained of pruritic papular urticarial
hypersensitivity was conrmed by positive lished and treating with antihistamine or eruptions that later extended to whole
epicutaneous skin test with DMSO to 5% body, dizziness, tachycardia and blackout.
10% and cutaneous biopsy consistent with
drug reaction. We applied a protocol of Table 1. For abstract 1609.
oral desensititazion to sorafenib, beginning Time Volume per Administered Cumulative
wih a dosage of 25 mg daily and increasing Step no. Solution no. Rate (ml/h) (min) step (ml) dose (mg) dose (mg)
every 3 days for getting the optimal dosage
1 1 2 15 0.50 0.02 0.02
of 800 mg in 21 days.
2 1 5 15 1.25 0.05 0.07
Results: No episodes of cutaneous reaction
3 1 10 15 2.50 0.10 0.17
occurred during or immediately after 4 1 20 15 5.00 0.20 0.37
desensitization. 3 weeks after completing 5 2 5 15 1.25 0.50 0.87
the desensititazion procotol we repeated 6 2 10 15 2.50 1.00 1.87
the epicutaneous skin tests with DMSO to 7 2 20 15 5.00 2.00 3.87
5% and 10% with negative result. 8 2 40 15 10.00 4.00 7.87
Conclusion: Epicutaneos skin test with 9 3 10 15 2.50 10.00 17.87
DMSO shows to be useful to conrm a 10 3 20 15 5.00 20.00 37.87
suspect of delayed hypersensitivity to So- 11 3 40 15 10.00 40.00 77.87
12 3 75 185 230.50 922.30 1000.00
rafenib also we demostrate than an oral
Allergy 66, Suppl. 94 (2011): 482642  2011 John Wiley & Sons A/S 607

The infusion was stopped; the patient was
treated with antihistamines and intrave-
nous steroids, and was observed for several
hours. After 1 month, given the success of
rituximab and the lack of response to other
drugs, the patient agreed to be re-treated
with rituximab using a desensitization pro-
tocol.
Results: The patient had received premedi-
cation with methylprednisone and phenir-
amine 30 min before the desensitization.
Three solutions in normal saline were pre-
pared and delivered in 12 consecutive steps
as shown in table 1, according to previous
literature.
2 weeks later, the subsequent rituximab
infusion was administered by means of
desensitization. The desensitization proce-
dure was successful and the patient subse-
quently tolerated.
Conclusion: Rapid desensitization is a
promising method for the delivery of ritux-
imab after immediate hypersensitivity reac-
tions to mAbs and should be considered
when there are no acceptable therapeutic
alternatives in patients with rheumatoid
arthritis.
1610
Oxaliplatin-induced fever as hypersensi-
tivity, and its management
Madrigal-Burgaleta, R1; Berges-Gimeno, M1; Guillen-
Ponce, C2; Sanz, M3; Angel-Pereira, D1; Rodrguez-
Vasquez, X1; Sanchez-Moreno, G1; Pena-Peloche, M1;
Olano-Rocha, M1; Alvarez-Cuesta, E1
1
Hospital Universitario Ramon y Cajal, Servicio de
Alergologa, Madrid, Spain; 2Hospital Universitario
Ramon y Cajal, Servicio de Oncologa Medica, Madrid,
Spain; 3Clnica Universitaria de Navarra, Departamento
de Alergologa e Inmunologa Clnica, Pamplona, Spain
Background: Oxaliplatin is known to be
involved in hypersensitivity reactions,
which are normally compatible with type I
hypersensitivity. Fever is not classically
considered to be a feature of immediate
hypersensitivity; however, cases of oxalipla-
tin-induced fever have been reported. The
pathophysiology of these fevers remains
unknown, but there has been speculation
on a possible immunological mechanism.
Fever reactions with oxaliplatin infusions
may entail important discomfort, and can
lead to therapy discontinuation before the
disease becomes refractory to treatment,
jeopardizing vital prognosis in many
patients.
Methods: We report the case of a patient
presenting with fever along with shivering,
general discomfort, and headache as the
manifestation of adverse reaction to an ox-
aliplatin-based chemotherapy metastatic
regime for his colorectal cancer. This prob-
lem was not observed during the rst infu-
sions, but appeared thirty minutes after
starting the tenth treatment. The set of
608
Poster Session 69 Desensitisation: an effective way of managing drug allergy
symptoms recurred in the next three
treatments despite intensied antipyretic,
corticosteroid, and antihistamine premedi-
cation. Infectious origin was ruled out in
every occasion. Treatment had to be sus-
pended, and the patient was referred to
our Desensitization Unit. Skin testing with
oxaliplatin (prick test with 5 mg/ml; intra-
dermal test with 0.5 mg/ml and 5 mg/ml),
and Basophil Activation Test (BAT) were
undergone in this patient (samples for
BAT were sent to Clnica Universitaria de
Navarra for their analysis). Medication
was administered by a rapid desensitization
to oxaliplatin, using the published stan-
dardized protocols of the Brigham and
Womens Hospital.
Results: Temporal tolerance was achieved
by rapid desensitization, and the patient
was able to receive his treatment, with
good clinical response to it. Skin testing
with oxaliplatin was negative. Basophil
Activation Test (BAT) showed a positive
response.
Conclusions: Tolerance to treatment can be
induced in patients presenting with fever as
the only manifestation of hypersensitivity
to oxaliplatin by the use of desensitization
protocols. Skin testing was unable to nd a
clear IgE involvement. BAT results may be
related to a possible participation of ba-
sophils in fevers induced by oxaliplatin in
this patient. The immunological mecha-
nism of these fevers is yet to be dened.
The implementation of BAT and desensiti-
zation protocols in the management of
these patients could be of inestimable clini-
cal use.
1611
Successful desensitisation with unfrac-
tioned heparin in a patient with immedi-
ate type hypersensitivity to low
molecular weight heparins but tolerant
to fondaparinux
Baccioglu Kavut, A1; Koca, E2
1
Ministry of Health, Erzurum Region Training and
Research Hospital, Department of Allergy and
Pulmonology, Erzurum, Turkey; 2Ministry of Health,
Erzurum Region Training and Research Hospital,
Department of Hematology, Erzurum, Turkey
Background: Heparin related anaphylaxis
like responses are rare, and the approach
to heparin allergic patient still remains
debatable. We aimed to diagnose heparin
allergy and to nd a way to treat a heparin
allergic patient with anticoagulants.
Methods: We performed skin and chal-
lenge tests with low-molecular-weight
(LMWH) and unfractioned heparin (UFH)
and a desensitization procedure with UFH.
Results: The patient with acute deep-vein-
thrombosis had urticaria, angioedema, and
bronchospasm just after the rst dose of
Allergy 66, Suppl. 94 (2011): 482642  2011 John Wiley & Sons A/S
enoxaparin. Then LMWH was stopped
and his symptoms were treated in the
emergency. Afterwards he referred to
Allergy Department and was desensitizated
with UFH and discharged with oral antico-
agulant-warfarin. In the following months,
he had some days with ineffective interna-
tional-normalised-ratio (INR) which might
result to recurrent vein thrombosis. So skin
tests were planned in order to nd alterna-
tive anticoagulant to enoxaparin for days
with ineffective INR. Enoxaparin and hep-
arin-sodium hypersensitivity was conrmed
by positive intradermal and challenge tests
respectively. Skin prick tests were negative
with other LMWHs including dalteparin,
nadroparin and tinzaparin while challange
by subcutaneously presented a positive
response with all. However the patient
didnt react to fondaparinux in skin and
challenge tests. Interestingly, the immediate
response to heparins was starting with
palmoplantar pruritis followed by bron-
chospazm, urticaria, angioedemea and
hypotension.
Conclusion: This a case of enoxaparin-
induced anaphylaxis like symptoms con-
rmed by skin tests whom was also found
to be sensitive to other LMWHs and UFH
which might be due to a cross-reactivity.
Palmoplantar pruritus after application of
heparins may be an early sign of immedi-
ate type hypersensitivity. Although he was
tolerated warfarin, fondaparinux and
desensitization with UFH was found to be
alternative therapies for emergent anti-
coagulation.
1612
Rapid oral desensitisation to isoniazid
Epozturk, K; Atayik, E; Abadoglu, O
Cumhuriyet University, Faculty of Medicine,
Department of Immunology and Allergic Diseases,
Sivas, Turkey
Background: Isoniazid is an essential drug
in the treatment and prevention of tuber-
culosis. Its use may lead to hypersensitivity
reactions. Some rapid desensitization pro-
tocols were previously reported.
Methods: We report the case of a 56-year-
old woman with no history of atopic dis-
ease. She has been followed for rheumatoid
arthritis for 6 years and was started to
receive adalimumab recently. Due to this
immunomodulatory treatment, she was
given 300 mg of isoniazid daily. Twelve
hours after the rst dose, she developed
urticarial skin lesions.
Results: Six weeks after the initial reaction,
skin tests with isoniazid were negative. In
the light of clinical manifestations, a desen-
sitization protocol was applied using
diluted forms of isoniazid pills.
Poster Session 69 Desensitisation: an effective way of managing drug allergy
The procedure proved to be successful
and the patient had no further reaction.
Table. For abstract 1612.
Time (min) Isoniazid (mg)
0 6.25
30 12.5
60 25
120 50
240 100
360 150
480 150
Next day 300
Conclusions: Rapid desensitization, a
method classically applied for IgE-medi-
ated hypersensitivity, can also be used for
the cases where the presence of drug-spe-
cic IgE was not demonstrated. This case
also showed that diluted oral pills can be
used for desensitization when other forms
were unavailable.
1613
Tolerance induction to clopidogrel
Gonzalez Ruiz, A; Gracia Bara, M; Hierro Santurino, B;
Moreno Rodilla, E; Ponce Guevara, V; Laffond Iges, E;
Munoz Bellido, F; Davila Gonzalez, I
Salamanca university Hospital, allergy department,
Salamanca, Spain
Background: The antiplatelet drug clopido-
grel is an oral thiopyridina derivate used in
dual antiplatelet therapy with aspirin for
the treatment of coronary diseases and
stent placement. Although clopidogrel is
well tolerated by most patients, hypersensi-
tivity reactions have been documented.
Methods: We describe a nonatopic, 76-
year-old female with a medical history of
diabetes, chronic renal failure and hyper-
tension who suffered a heart attack in
Allergy 66, Suppl. 94 (2011): 482642  2011 John Wiley & Sons A/S
October 2010. After a coronary catheteri-
zation a stent was implanted in the right
coronary artery and treatment with clopi-
dogrel 75 mg plus aspirin 100 mg/day was
initiated. Thirteen days later she developed
Reaction
generalized rash and pruritus. The cardiol-
- ogist discontinued clopidogrel, the rash dis-
- appeared in 8 days. After 6 days the
- patient suffered a second heart attack.
-
Another catheterization was performed
-
and the cardiologist contacted the allergy
-
- department by the need for dual antiplat-
- elet therapy. An alternative thienopyridin,
that may be a substituted for clopidogrel is
ticlopidine; however side effects have been
reported and published a recurrence risk of
27%. Other alternatives are prasugrel or ti-
cagrelor, but cardiologists preferred to use
treatment with clopidogrel.
Results: We choose a rapid oral desensiti-
zation/tolerance induction in the intensive
care unit. Doses were administered follow-
ing a protocol of 3.5 h, reaching an acu-
mulative dose 142.2 mg without any
reaction. Treatment with clopidogrel was
continued until now without further reac-
tions.
Conclusion: We present a case of tolerance
induction to clopidogrel. In these patients,
due to the high risk of new coronary dis-
ease, it is important to conduct early toler-
ance induction.
1614
Desensitisation to thalidomide
Avila Castellano, R; Cimbollek, S; Gonzalez-Quevedo, T;
Pinero, M; Ortega Camarero, M; Quiralte, J
Hospital Universitario Virgen del Rocio, U.G.C.
Intercentros Valme/Rocio de Alergologia, Sevilla, Spain
Background: Thalidomide due to its anti-
angiogenic mechanism of action is a thera-
peutic option for patients with frequently
gastrointestinal blood loss due to angiodys-
plasias who no longer tolerate conventional
and invasive procedures due to their physi-
cal conditions. We report a case of thalido-
mide hypersensitive in a patient with
cirrhosis and diffuse colonic angiodysplasia
pending hepatic transplant successfully
desensitized.
Methods: A 65-year-old man diagnosed of
enolic cirrhosis and diffuse colonic angi-
odysplasia was treated with oral thalido-
mide 200 mg daily during 4 months with
adequate tolerance. The patient referred
generalized pruritus and edema in legs
when doses was increased to 300 mg. The
treatment was discontinued and some
months later thalidomide 100 mg o.d was
reintroduced. Patient developed immediate
generalized urticaria the second day. Tha-
lidomide was required for optimal angi-
odysplasia treatment until transplant.
Risks and benets were discussed with the
patients and informed consent was
obtained. A desensitization protocol was
designed, administering increasing doses of
different dilutions of thalidomide every
30 min. First day: 1 mcg, 2 mcg, 3 mcg,
4 mcg. Second day: 0.1 mg, 0.2 mg,
0.3 mg, 0.4 mg. Third day: 1 mg, 2 mg,
4 mg, 8 mg, 16 mg, 32 mg, 40 mg. Fourth
day: 100 mg. Premedication with oral
hidroxicine 2 mg/ 8 h and prednisone
15 mg was administered.
Results: Desensitization protocol was
uneventful. Prick and patch test were per-
formed with negative results.
Conclusions: Desensitization appears to be
a safe and valid therapeutic option in
patients with hypersensitivity reaction to
thalidomide if no better treatment option
is available.
609

Understanding atopic dermatitis
1615
Serum levels of sCD30 correlate with age
and with disease extent and severity in
patients with atopic dermatitis
Rosinska-Wieckowicz, A; Czarnecka-Operacz, M; Silny,
W
Poznan University of Medical Sciences, Department of
Dermatology, Poznan, Poland
Background: Soluble immune receptors
(SIRs) have been considered as biomarkers
in patients with atopic dermatitis (AD).
The aim of the study was to investigate the
usefulness of evaluation of serum levels of
the soluble form of CD30 (sCD30) to mon-
itor disease extent an severity in patients
with AD.
Methods: Hundred and two patients with
AD and two control groups: 20-allergic
rhinitis (AR), 20-healthy individuals (HI).
Following procedures were performed:
clinical assessment of AD group (SCO-
RAD, W-AZS index), skin prick tests
(SPT) with food and airborne allergens
(HI, AR and in 64/102 AD patients
>3 years), furthermore evaluation of
serum levels of total IgE (tIgE), antigen -
specic asIgE (asIgE) ImmunoCAP), and
sCD30 (ELISA) (HI, AR, AD).
Results: The mean age of the investigated
population was 9.7 9.1 year (75.5%
of patients <18 years, and 45.5% > 18
years). The mean SCORAD was 42.9
24.5 while the mean W-AZS was 39.4
29.8. Based on patients history and test
results allergic type of AD (aAD) was
diagnosed in 62% and non-allergic type of
AD (naAD) in 32% of the AD group.
Serum levels of sCD30 were signicantly
higher in patients with AD in comparison
with patients with AR (P = 0) and HI
(P = 0). Signicant negative correlation
was observed between disease extent and
severity and serum levels of sCD30 (SCO-
RAD: P = 0.009, Spearman ratio - rs:
)0.257, W-AZS: P = 0.002, RS: )0.307)
in the whole investigated AD population.
Furthermore, signicant negative correla-
tion was also observed between serum lev-
els of sCD30 and recorded W-AZS values
in aAD group (P = 0.043, RS: )0.239).
Moreover, we observed a negative correla-
tion between the age of AD patients and
serum levels of sCD30, both in the whole
investigated group (P = 0, RS: )0.782) as
well as in the aAD group (P = 0, RS:
610
Poster Session 70
)0.757) and naAD group (P = 0, RS:
)0.708).
Conclusion: Serum levels of sCD30 in
patients with AD were signicantly age-
dependent, and correlated negatively with
disease severity. As it was proved that
sCD30 correlates inversely with age even in
healthy individuals, it may not be consid-
ered an applicable parameter to monitor
disease extent and severity in patients with
AD.
1616
Serum levels of interleukin 13 (IL-13) and
soluble interleukin 2 receptor (sIL-2Ra)
were not applicable parameters to moni-
tor disease extent and severity in
patients with atopic dermatitis
Rosinska-Wieckowicz, A; Czarnecka-Operacz, M; Silny,
W
Poznan University of Medical Sciences, Department of
Dermatology, Poznan, Poland
Background: The aim of the study was to
investigate the usefulness of evaluation of
serum levels of IL-13 and sRaIL-2 to mon-
itor disease extent an severity in patients
with atopic dermatitis (AD).
Methods: Hundred and two patients with
AD and two control groups: 20-allergic
rhinitis (AR), 20-healthy individuals (HI).
Following procedures were performed:
clinical assessment of disease extent and
severity in the AD group (SCORAD, W-
AZS index), skin prick tests (SPT) with
food and airborne allergens (HI, AR and
in 64/102 AD patients>3 years), further-
more evaluation of serum levels of total
IgE (tIgE), antigen - specic asIgE (asIgE)
ImmunoCAP), and IL-13 and sIL-2Ra
(ELISA)(HI, AR, AD).
Results: Based on patients history and test
results allergic type of AD (aAD) was
diagnosed in 62% and non-allergic type of
AD (na-AD) in 32% of the AD group.
Serum levels of IL-13 were signicantly
higher in patients with AD in comparison
with patients with AR (P = 0.002) and HI
(P = 0.0019). Serum levels of sIL-2Ra
were also signicantly higher in patients
with AD in comparison with patients with
AR (P = 0) and HI (P = 0). Signicant
positive correlation was found between
serum levels of tIgE and W-AZS and SCO-
RAD values (P = 0, R Spearman - RS:
Allergy 66, Suppl. 94 (2011): 482642  2011 John Wiley & Sons A/S
0.658; P = 0, RS )0.649). However no
correlation was found between disease
extent and severity (W-AZS) and serum
levels of IL-13 in the whole investigated
group of patients with AD (P = 0.394,
RS: )0.085) as well as in the aAD group
(P = 0.141, RS: )0.175) and naAD group
(P = 0.459, RS: )0.140). Furthermore no
correlation was found between disease
extent and severity (SCORAD) and serum
levels of IL-13 in the whole investigated
group of patients with atopic dermatitis
(P = 0.695, RS: )0.039) as well as in the
aAD group (P = 0.114, RS: )0.340) and
naAD (P = 0.904, RS: )0.023). Signicant
negative correlation was observed between
disease extent and severity and serum levels
of sIL-2Ra (W-AZS: P = 0.004, RS:
)0.199, SCORAD: P = 0.049, RS:
)0.196), while in aAD and nAD groups
analogous correlations were not found.
Conclusions: Evaluation of serum levels of
IL-13 and sIL-2Ra were not useful param-
eters to monitor disease extent and severity
in patients with atopic dermatitis.
1617
Elevated serum thymic stromal lympho-
poietin in infants and toddlers with
atopy
Tepper, R; Zhou, B; Yao, W; Kaplan, M
Indiana University, Pediatrics, Indianapolis, United
States
Background: The development of atopic
dermatitis and asthma is associated with
the presence of thymic stromal lympho-
poietin (TSLP). Recent animal studies sug-
gest that TSLP may be the underlying
factor driving the progression from atopic
dermatitis to asthma; however, data on
TSLP is very limited in humans, particu-
larly early in life.
Methods: Infants with eczema were
recruited prior to any wheezy respiratory
illnesses and followed-up 1 year later. Sub-
jects were characterized as atopic based
upon serum IgE. We selected a subset of
serum samples obtained upon entry of
study (rst year) and at follow-up (second
year) and measured TSLP by Elisa.
Results: Although the disease severity of
dermatitis was not signicantly different
between atopic (n = 12) and non-atopic
(n = 16) infants at rst year when they
Poster Session 70 Understanding atopic dermatitis
were recruited into the study, the atopic
dermatitis infants had signicantly higher
serum TSLP concentration. When they
reached 2 years old, the atopic subjects
had more severe dermatitis and exhibited
even higher serum TSLP concentration
compared with non-atopic subjects.
Conclusion: Our ndings suggest that
TSLP is elevated in atopic subjects early in
life.
1618
Interleukin-9 in patients with cutaneous
diseases
De Amici, M1; Marseglia, G2; Berardi, L3; Legoratto, S3;
Giunta, V1; Mantegna, G1; Licari, A4; Vignini, M3
1
Foundation IRCCS Policlinic San Matteo, Hospital
Pediatric Clinic, Pavia, Italy; 2Foundation IRCCS
Policlinic San Matteo, University Pediatric Department,
Pavia, Italy; 3Foundation IRCCS Policlinic San Matteo,
Dermatology Clinic, Pavia, Italy; 4Foundation IRCCS
Policlinic San Matteo, Neonatal Intensive Care Unit,
Pavia, Italy
Background: T helper (Th) cells seem to be
the main source of IL-9 which is an impor-
tant cytokine in allergic diseases such as
asthma and atopic dermatitis. The aim of
this study was to evaluate serum IL-9 lev-
els in patients with cutaneous hypersensi-
tivity diseases, including atopic dermatitis,
contact dermatitis and urticaria. A group
of healthy subjects was included as a con-
trol.
Methods: The study included 47 patients
(25 females, 22 males, median age 38 years)
with cutaneous hypersensitivity diseases,
including atopic dermatitis, contact derma-
titis and urticaria, and 24 healthy subjects
(12 females, 12 males, median age
44 years). Cutaneous disorders may be
classied (acute/chronic) on the basis of
symptom duration. Thirty-one patients
(66%) showed chronic symptoms; sixteen
patients (34%) acute symptoms. The
human interleukin 9 (Human IL-9 ELISA
Ready-SET-Go!, eBioscience, Rome, Italy)
reagent kit contains the necessary reagents,
buffers and diluents for performing quanti-
tative enzyme linked immunosorbent
assays (ELISA). Statistical analysis was
performed using the statistical software
package Medcalc 9 (Frank Schoonjans,
BE). Descriptive statistics were rst per-
formed and quantitative parameters were
reported as medians (md) and 25th-75th
percentiles (IQR) due to the skewed distri-
bution. The non parametric Wilcoxon test
was performed to evaluate differences
between groups. A P-value 0.05 was con-
sidered statistically signicant.
Results: The comparison between patients
with cutaneous diseases and healthy sub-
jects showed a signicant (P = 0.0093) dif-
ference in median serum IL-9 levels
(patients: md: 13.32 pg/ml; IQR: 8.79
Allergy 66, Suppl. 94 (2011): 482642  2011 John Wiley & Sons A/S
17.44 pg/ml. Healthy subjects: md: 6.31 pg/
ml; IQR: 0.0922.01 pg/ml). The compari-
son between patients with acute symptoms
and chronic symptoms showed a signicant
(P = 0.0389) difference in median serum
IL-9 levels (acute symptoms: md: 16.79 pg/
ml; IQR: 10.9819.55 pg/ml. Chronic
symptoms: md: 11.11 pg/ml; IQR: 8.14
15.15 pg/ml).
Conclusions: The present preliminary nd-
ings support the supposition that serum
IL-9 levels are increased in patients with
cutaneous diseases. However, further stud-
ies, conducted on larger populations,
should address the role of symptom dura-
tion in modulating serum IL-9 levels in
patients.
1619
Expression of thymic stromal lympho-
poietin in canine atopic dermatitis
Klukowska-Rotzler, J1; Chervet, L2; Muller, E2; Roosje,
P3; Marti, E4; Janda, J4
1
Vetsuisse Faculty, University of Bern, Institute for
Genetics, DCR-VPH, Bern, Switzerland; 2Vetsuisse
Faculty, University of Bern, Institute for Animal
Pathology, Bern, Switzerland; 3Vetsuisse Faculty,
University of Bern, Dermatology Unit, Bern,
Switzerland; 4Vetsuisse Faculty, University of Bern,
Clinical Immunology, DCR-VPH, Bern, Switzerland
Background: Canine atopic dermatitis
(cAD) is a highly pruritic allergic dermati-
tis caused by reactions to environmental
allergens, especially house dust mites. T
helper 2 (Th2) differentiation and allergen-
specic IgE antibodies have been impli-
cated in the pathogenesis of cAD, but the
mechanisms involved in the induction of
Th2 responses are not known. In mice and
humans, thymic stromal lymphopoietin
(TSLP) has been shown to induce Th2 dif-
ferentiation and allergic dermatitis. The
aim of the present study was to identify
the canine TSLP homolog and characterize
expression of canine TSLP in the skin dogs
affected by cAD.
Methods: To clone the canine TSLP
cDNA, sequences homologous with human
TSLP were identied in the canine genome
using BLASTN. We cloned and sequenced
a partial coding sequence and established
quantitative RT-PCR to assess the expres-
sion of canine TSLP in cultured canine
keratinocytes and in skin biopsies from le-
sional and non-lesional skin of 12 cAD-
affected dogs and six healthy control dogs.
Results: Nucleotide sequence of canine
TSLP is organized in four exons and shows
homology with other species (human 70%
identity, equine 73% identity). Deduced
amino acid sequence shows 60.8% and
59.9% identity with human and equine
TSLP proteins. Quantitative RT-PCR
showed that the expression of TSLP was
higher in both lesional and non-lesional
skin of cAD-affected dogs compared to
healthy control dogs (P < 0.05), whereas
no signicant difference was found between
lesional and non-lesional samples. Since
keratinocytes have been proposed as
potential sources of TSLP, we sought to
determine whether and under which condi-
tions canine keratinocytes express TSLP.
Stimulation of primary canine keratino-
cytes for 18 h with allergen (D. farinae
extract) and with toll-like receptor ligands
LPS (TLR4) and polyI:C (TLR3) resulted
in increased expression of TSLP, whereas
stimulation with Pam3Cys (TLR2) or imiq-
uimod (TLR7) did not increase TSLP
expression.
Conclusions: In analogy to humans and
mice, our nding of increased TSLP
expression in the skin of dogs with atopic
dermatitis suggests that TSLP may also
play an important role in the pathogenesis
of cAD. Keratinocytes were identied as
potential sources of TSLP, as they
responded to stimulation with allergen
extract by increased TSLP expression. Fur-
ther studies should elucidate the function
of TSLP in cAD and explore its potential
as a therapy target.
1620
Extracellular vesicles derived from Staph-
ylococcus aureus induce atopic dermati-
tis-like skin inflammation in mice
Hong, S1; Kim, M1; Lee, E1; Yang, J2; Kim, Y1; Jeon, S1;
Kim, Y1
1
POSTECH, Republic of Korea; 2Republic of Korea
Background: Recently, we found that
Staphylococcus aureus produces extracellu-
lar vesicles (EV) that contain pathogenic
proteins. Although S. aureus infection has
been linked with atopic dermatitis (AD),
the identities of the causative agents are
controversial. We evaluated whether S.
aureus-derived EV are causally related to
the pathogenesis of AD.
Methods: EV were isolated by the ultracen-
trifugation of S. aureus culture media. The
EV were applied three times per week to
tape-stripped mouse skin. Inammation
and immune dysfunction were evaluated
48 h after the nal application. EV-specic
IgE levels were measured by ELISA in AD
patients and healthy subjects.
Results: The in vitro application of
S. aureus EV increased the production of
pro-inammatory mediators, including
TNF-alpha, IL-6, eotaxin, and MIP-
1alpha, by mouse dermal broblasts. The
in vivo application of S. aureus EV after
tape stripping caused epidermal thickening
with inltration of the dermis by mast cells
and eosinophils in mouse skin. These
changes were associated with the enhanced
611

cutaneous production of IL-4, IL-5, IFN-
gamma, and IL-17. Interestingly, the serum
levels of S. aureus EV-specic IgE were sig-
nicantly increased in AD patients relative
to healthy subjects.
Conclusions: These results indicate that S.
aureus EV induce AD-like inammation in
the skin and that S. aureus-derived EV are
a novel diagnostic and therapeutic target
for the control of AD.
1621
Histocompatibility antigens and the
relative risk of atopic dermatitis
Drannik, G1; Kurchenko, A1; DuBuske, L2
1 tinct succession of atopic diseases and are
National Medical University, Kiev, Ukraine;
2
Immunology Research Institute of New England,
Gardner, MA, United States
Background: Atopic dermatitis (AD) is a
systemic allergic disease that has a 1020%
incidence in economically developed coun-
tries. 10% to 40% of adult AD patients
have IgE independent disease with no evi-
dence of allergen sensitization, contrary to
the IgE-dependent form, which is charac-
terized by higher IgE levels and sensitiza-
tion to allergens. This study assesses the
impact of HLA histocompatible antigens
on the relative risk for both forms of AD.
Methods: The distribution of HLA-A, B,
and DR antigens in 160 adult patients suf-
fering from IgE-dependent (80 persons
Group I) and IgE-independent (80 persons
Group 2) forms of atopic dermatitis
(AD) was analyzed. The HLA antigens
were assessed using a standard microlym-
phocytotoxic test and special panels of
anti-HLA serums (20 antigens of locus A,
31 B and 9 - DR). The control group
consisted of 250 healthy donors from Kyiv.
Results: Patients from Group 1 had a rel-
ative risk for AD to develop that was sta-
tistically greater in carriers with HLA-A24
expression (RR = 2.70 with s = 0.12).
The investigation of class II loci showed
that attributive risk was available for
development of IgE-dependent AD for
individuals having antigens B7
(RR = 2.47, s = 0.13), B22 (RR = 2.31,
s = 0.10), B27 (RR = 2.55, s = 0.12),
and DR2 (RR = 2.50, s = 0.11). In
patients with the IgE-independent AD, no
signicant associations were seen with
HLA expression.
Conclusion: Increased relative risk for the
development of IgE-dependent AD was
seen associated with carriers of HLA-anti-
gens including A24, B7, B22, B27, and
DR2. The absence of the signicant HLA
associations in the patients of IgE indepen-
dent AD suggests an absence of connection
with specic HLA-phenotypes.
612
Poster Session 70 Understanding atopic dermatitis
1622
In a murine model of allergic skin inflam-
mation, an ongoing allergic skin inflam-
mation does not facilitate priming
towards new unrelated antigens
Mahapatra, S1; Robinson, E2; Herrick, C2; Dittrich, A3
1
Hannover Medical School, Department of Pediatric
Pneumology, Allergology & Neonatology, Hannover,
Germany; 2Yale University School of Medicine,
Department of Dermatology, New Haven, United
States; 3Hannover Medical School, Department of
Pediatric Pneumology, Neonatology & Allergology,
Hannover, Germany
Background: Atopic dermatitis (AD) is
often the rst clinical manifestation of the
atopic march. However, the reason why
individuals with AD often develop a dis-
susceptible to polysensitization towards dif-
ferent innocuous antigens is still unclear.
We have previously shown that a Th2-
polarized pulmonary inammation can
facilitate priming to new, unrelated anti-
gens via the airways, a process which we
call collateral priming. We were hence
interested whether Th2-polarized allergic
skin inammation can also facilitate prim-
ing towards new antigens, similar to what
we observed in the lung.
Methods: To address the above hypothesis,
we established two different murine models
of sensitization via the skin and choose
Ovalbumin (OVA) & Keyhole limpet
hemocyanin (KLH) as two unrelated anti-
gens. In one model BALB/c mice were
initially sensitized towards OVA epicutane-
ously. The second model was based on
OVA sensitization via an OVA/alum i.p.
injection. For the skin models, mice are
challenged epicutaneously with OVA
together with sub-immunological doses of
KLH. After 2 weeks they are re-challenged
epicutaneously with KLH and after 3 days
we analyzed specic antibody responses in
the serum, proliferation and cytokine pro-
le of the draining lymph nodes and skin
inammation of these mice to assess
whether priming towards the secondary
antigen KLH has occurred. These models
were developed in analogy to a lung model
of collateral priming where facilitated
priming occurs to a secondary antigen
when it is given in sub-immunological dose
along with a robust dose of a potent
antigen.
Results: In contrast to the lung model, we
did not observe signicant levels of KLH
specic IgG1 and IgE in the mice which
had received OVA and sub-immunological
dose of KLH epicutaneously during rst
challenge followed by the secondary chal-
lenge of KLH. Histological examination of
the skin of these mice also didnt show sig-
nicant inammation compared to nega-
tive control. Lymph node proliferation &
Allergy 66, Suppl. 94 (2011): 482642  2011 John Wiley & Sons A/S
cytokine secretion also support that no
priming was seen to KLH in these mice.
Conclusions: We conclude that, in contrast
to an ongoing pulmonary inammation, an
ongoing allergic skin inammation doesnt
facilitate priming to a secondary, unrelated
antigen suggesting that the immunological
mechanisms governing skin inammation
differ profoundly from those applying to
lung inammation.
1623
Cytokine gene expression in the skin and
peripheral blood in atopic dermatitis
patients and healthy donors
Filimonova, T1; Elisyutina, O2; Fedenko, E2; Boldyreva,
M2
1
Institut Immunology, Skin Allergy Department,
Moscow, Russian Federation; 2Russian Federation
Background: Investigation of immune
response in the skin and peripheral blood
of atopic dermatitis (AD) patients and
healthy donors.
Materials and methods: Fourty adult sub-
jects, aged from 17 to 45 years, were
included in the study and separated into 2
groups. Group 1 made 20 patients with
lasting recurrent AD, aged from 16 to
45 years, of whom four patients had severe
AD, 14 patients moderate AD, two
patients mild AD. Group 2 made 20
adult healthy donors without signs of
atopy, aged from 19 to 32 years. Materials
for the investigation were skin biopsy and
peripheral blood samples of AD patients
and healthy donors. Nytokines gene
expression were dened with polymerase
chain reaction in real time (DNA-technol-
ogy, Russia). The level of gene expression
of these cytokines were measured in stan-
dard units.
Results: There was statistically signicant
increase level of gene expression IL2r
(P < 0.001), IL5 (P < 0.003), IL6 (P <
0.002), IL8 (P < 0.000003), IL12B (P <
0.003), IL10 (P < 0.00006), IL23
(P < 0.042), IL29 (P < 0.006), gamma-
interferon (P < 0.026), TGFB (P <
0.0006) in the skin biopsy samples of AD
patients compared with healthy donors.
There was statistically signicant increase
level of gene expression IL5 (P < 0.049),
IL12B (P < 0.011), gamma-interferon
(P < 0.004) in peripheral blood of AD
patients compared with healthy donors.
Both in the skin and in the peripheral
blood there was statistically signicant
increase level of gene expression IL12B,
IL5, gamma-interferon in AD patients
compared with healthy donors.
Conclusion: The study has shown the activ-
ity IL-2r, IL-5, IL-8, IL-12B, IL-23, IL-29,
gamma-interferon, TGFB, which are mark-
ers of chronic inammation and the
Poster Session 70 Understanding atopic dermatitis
predominance of Th1-immune response in
patients with lasting recurrent AD.
1624
Analysis of filaggrin loss-of-function
mutations in atopic dermatitis patients
and populations of the Volga-Ural region
of Russia
Gimalova, G1; Karunas, A1; Fedorova, Y1; Gumennaya,
E2; Imangulova, M1; Biktasheva, A3; Levashova, S3;
Etkina, E3; Khusnutdinova, E1
1
Institute of Biochemistry and Genetics, Ufa Science
Centre of RAS, Genomics Department, Ufa, Russian
Federation; 2Dermatovenerologic dispensary of
Bashkortostan Republic, Ufa, Russian Federation;
3
Bashkir State Medical University, Ufa, Russian
Federation
Atopic dermatitis is a chronic inamma-
tory skin disorder that typically presents in
early childhood and may continue or recur
later. AD is a complex disease involving
gene-gene and gene-environment interac-
tions. Recent studies have suggested that a
primary epidermal barrier defect may play
a crucial role in the development of atopic
dermatitis. The laggrin protein is a key
component of stratum corneum involved in
skin barrier function. Filaggrin loss-of-
function mutations lead to complete loss of
laggrin expression, which causes exces-
sively dry skin and impaired skin barrier
function. Mutations in the prolaggrin
gene (FLG) have been identied as a
strong predisposing factor for atopic dis-
eases. In this study, we have screened two
FLG loss-of-function mutations (2282del4
and R501X) in AD patients, control indi-
viduals and populations of the Volga-Ural
region of Russia. The AD group consisted
of 359 AD patients with different ethnic
origins (131 Russians, 76 Tatars, 29 Bash-
kirs). The control group included 266 non-
atopic individuals (85 Russians, 57 Tatars
and 50 Bashkirs). Examined populations
consisted of 102 Russians, 106 Tatars and
282 Bashkirs. Genome DNA was excluded
by phenol-chloroform extraction from
peripheral blood. Mutations detection was
performed by PCR-RFLP and sequencing.
We have determined a strong association
of FLG mutation 2282del4 with AD in
Russians, Tatars and Bashkirs. The allelic
frequency of the 2282del4 mutation was
5.99% in combined AD group, 6.49% in
patients of Russian ethnic origin, 8.55% in
Tatars and 10.34% in Bashkirs. In control
group, the frequency of 2282del4 was sig-
nicantly lower: 1.88% in combined group,
1.18% in Russians, 0.88% in Tatars and
2.44% in Bashkirs, accordingly. We have
evaluated the frequency of heterozygous
carriers of the 2282del4 mutation in three
populations: Russians, Tatars and Bash-
kirs. This mutation was detected with fol-
lowing frequencies - 1.96% in Russians,
Allergy 66, Suppl. 94 (2011): 482642  2011 John Wiley & Sons A/S
1.42% in Tatars and 3.55% in Bashkirs.
The R501X mutation was found in AD
patients with low frequency (0.59%) and
was absent in control individuals. The data
of this study revealed that the FLG muta-
tion 2282del4 is an important risk factor of
atopic dermatitis in the Volga-Ural region
of Russia.
1625
No systemic defect in dendritic cells from
individuals with early-onset psoriasis
Kimber, I1; Shaw, F1; Begum, R1; Griffiths, C2; Dearman,
R1
1
University of Manchester, Faculty of Life Sciences,
Manchester, United Kingdom; 2University of
Manchester, Faculty of Medicine and Health Sciences,
Manchester, United Kingdom
Background: We have demonstrated previ-
ously that the migration of epidermal Lan-
gerhans cells (LC) in response to the
cytokines interleukin-1 beta (IL-1b) and
tumour necrosis factor-alpha (TNF-a) is
impaired in uninvolved skin of individuals
with early-onset (age of onset <40y) psori-
asis. We have now investigated whether
this defect represents an underlying sys-
temic defect in dendritic cell (DC) function
in these individuals using blood derived
monocytes differentiated into LC-like cells
(mLC).
Methods: CD14+ cells were isolated from
the blood of both healthy volunteers and
individuals with psoriasis and cultured for
5 days in the presence of a cytokine cock-
tail to promote their differentiation into
mLC. Cells were stimulated for 24 h with
TNF-a, IL-1b (both 100 ng/ml) or with
medium alone. Membrane marker expres-
sion was quantied by ow cytometry and
ability to migrate to CCL19 was assessed
using a Transwell migration assay. The
cytokine and chemokine content of super-
natants was analysed by cytokine bead
array.
Results: Similar numbers of CD14+ cells
were isolated from both groups and in
each case cells acquired a LC-like pheno-
type with high expression of CD1a and
MHC Class II with culture. Stimulation
with TNF-a and IL-1b increased expres-
sion of activation markers (CD86 and
CD54) to the same extent in both groups.
Mature mLCs (those cultured with IL-
1bn or TNF-a) migrated to CCL19, more
vigorously when activated with the former,
and there was no difference in migratory
capacity between healthy volunteers and
individuals with early onset psoriasis.
Equivalent levels of basal and cytokine-
stimulated secreted cytokines and chemo-
kines were recorded also, with, for exam-
ple, up-regulation of IP-10, MIP-1 and
TNF-a recorded following activation with
IL-1b.
Conclusions: These data demonstrate that
there is not a generalised functional defect
in the ability of all DCs to respond to acti-
vation in individuals with early- onset pso-
riasis. This suggests that the lack of
migration observed in vivo is due to either
an epidermal LC-specic effect, or to the
cutaneous microenvironment.
1626
Pemphigus vulgaris autoantibodies
recognise neither a7 not a9 subunits of
nicotinic acetylcholine receptor
Tatsiy, O1; Melnikova, V2; Kryukova, E3;
Svirshchevskaya, E3
1
Shemyakin-Ovchinnikov Institute of Bioorganic
Chemistry, RAS, Moscow, Russian Federation; 2Koltzov
institute of Developmental Biology, RAS, Moscow
Russian Federation; 3Shemyakin-Ovchinnikov Institute
of Bioorganic Chemistry, RAS, Moscow Russian
Federation
Background: Nicotinic acetylcholine recep-
tors (nACR) belong to the superfamily of
ion channels which are opened by specic
ligands and which regulate membrane cur-
rent in muscle and neuronal cells. There
are two types of nACR: muscle and neuro-
nal ones, the variety of which is a result of
a2-a10 and b2-b4-subunit combinations.
There are data that autoantibodies to
nACR and especially to a9-subunit of
nACR play a role in pemphigus vulgaris
(PV). PV is a severe autoimmune dermato-
sis which is induced by autoantibodies to
keratinocyte antigens. The binding of
autoantibodies to keratinocytes induces
epidermal acantholysis. The major autoan-
tigen in PV is desmoglein 3, however
autoantibodies to nACR can also be of
importance. The aim of this work was the
identication of autoantibodies to a9 and
a7 nACR in PV serum.
Methods: To identify specic subunits of
nACR a selective inhibitor of a7, a9
nACR - a-bungarotoxin (a-BT), labeled by
AlexaFluor555 was used. As a positive
control of a-BT binding to a9 nACR we
used cryosections of rat inner ear to visual-
ize in organ of Corti type II hair cells
which are known to express a9 nACR. For
a7 nACR binding we used human cells
THP-1 which were shown to express a7
subunit of nACR. PV IgG were puried
from sera pooled from 25 PV patients.
Both cryosections and live THP-1 cells
were stained with a-BT, PV or normal
human IgG, xed and analyzed using con-
focal microscopy.
Results: We demonstrated binding of a-BT
with rat cochlea which was blocked by a-
cobratoxin but not by a-neurotoxin II
showing a selective binding of a-BT. The
same inhibition was found when we stained
613

THP-1 cells with a-BT. Thus, a-BT binds
specically to a7 and a9 nACR. To study
the existence of autoantibodies to a7 and
a9 nACR in PV serum cryosections or
THP-1 cells were co-incubated with PV or
normal human IgG and a-BT. We found
no co-localization of staining and no dif-
ference between PV and normal human
IgG staining neither in type II hair cells in
organ of Corti nor in THP-1 cells.
Conclusion: Thus, we concluded that
neither a7 nor a9 nACR are targets for
autoantibodies in PV.
1627
Serum levels of angiogenin, IL-4, IL-6,
IL-10, IL-17 and TNF-alpha in patients
with mastocytosis
Guler Ozden, M1; Summer, B2; Chen, Y3; Rueff, F4
1 controls (no statistical signicant differ-
Ondokuz Mayys University, Dermatology, Samsun,
Turkey; 2AllergieZentrum, Ludwig-Maximilian
University, Department of Dermatology and
Allergology, Munich, Germany; 3National Defense
Medical Center, Taichung, Taiwan; 4Ludwig-Maximilian
University, AllergieZentrum, Department of
Dermatology and Allergology, Munich, Germany
Background: Angiogenin is a potent angio-
genic protein and seems to play a role in a
variety of angiogenesis-dependent patho-
logical conditions. Recently, it has been
shown that angiogenesis is also increased
in bone marrow of patients with systemic
mastocytosis (SM). Human mast cells
secrete angiogenin and cytokines (including
interleukin (IL)-3, IL-4 and IL-10) in
response to a variety of stimuli. These
mediators are thought to play a role in
mastocytosis and its clinical complications
(osteoporosis).
Aim: We examined the association of SM
with angiogenin and a variety of cytokines
in the serum.
Material and Methods: The serum samples
of 20 patients with established SM (40%
male, 3680 years old) were analyzed. Ten
patients were suffering from SM; two
patients had a cutaneous mastocytosis
(CM), and eight patients presented with
CM, which was combined with one or two
minor diagnostic criteria for SM. Patients
were classied according to WHO criteria.
614
Poster Session 70 Understanding atopic dermatitis
In total, eight (40%) patients with masto-
cytosis were suffering from osteoporosis
and ten (50%) patients had a history of
anaphylaxis. Sera of ten patients with
chronic urticaria, and of ten healthy volun-
teers served as controls. The serum samples
had been frozen at )70C until measure-
ment. Cytokines (IL-4, 6, 10, 17, 1-beta
and tumor necrosis factor alpha) were ana-
lyzed by Multiplex Cytometric Bead Array
(BD Biosciences). Serum tryptase was mea-
sured by ImmunoCAP (Phadia).
Results: Serum tryptase concentration in
patients with SM was increased above nor-
mal in patients with SM (mean value
33.4 30.0 lg/l). There were no detect-
able levels of IL-4, 6, 10, 17, 1-beta and
TNF-alpha in the serum of patients with
SM and controls. Angiogenin was present
in all plasma samples of all patients and
ence). There was no correlation between
angiogenin levels and serum tryptase con-
centration, presence of clinical symptoms,
or extent of mastocytosis.
Conclusions: Immunologic activity of
serum interleukins does not correlate with
secondary complications of mastocytosis.
Presumably, the activity of local cells (bone
marrow) is more important for mediating
secondary pathological changes.
1628
Angelicae dahuricae radix inhibits dust
mite extract-induced atopic
dermatitis-like skin lesions in NC/Nga
mice
Hoyoung, L; Hyekyung, H; Jun Kyung, L; Mee-Young, L;
Chang-Seob, S; Hyeunkyoo, S
Korea Institute of Oriental Medicine, Herbal Medicine
EBM Research Center, Daejeon, Republic of Korea
Background: Angelicae Dahuricae Radix
(AR) is the dried root of Angelica dahurica
(FISCH.) BENTH. et HOOKER F (Umbellife-
rae). AR has been used for the treatment
of colds, headaches, toothaches, coryza,
and psoriasis in Korean traditional medi-
cine. We examined whether Angelicae
Dahuricae Radix (AR) suppresses the
development of atopic dermatitis (AD)-like
Allergy 66, Suppl. 94 (2011): 482642  2011 John Wiley & Sons A/S
skin lesions induced by Dermatophagoides
farinae in NC/Nga mice.
Methods: To investigate the effect of AR
in AD induction, we measured the AD
severity score, measured plasma levels of
IgE and histamine, and performed histo-
logical analysis in AD-induced NC/Nga
mice. Ten milligrams of AR extract was
applied daily for 4 weeks to the AD-like
skin lesions. We also conrmed the anti-
inammatory effects of AR by measuring
thymus and activation-regulated chemoki-
ne (TARC/CCL17) protein production and
mRNA levels of TARC and macrophage-
derived chemokine (MDC/CCL22) in
TNF-alpha/IFN-gamma-treated HaCaT
cells. The effects of AR on NO production
were examined in LPS-induced RAW 264.7
cells.
Results: Both the AR extract and 0.1% ta-
crolimus (positive control immunosuppres-
sant) suppressed the development of AD-
like skin lesions and reduced dermatitis
scores of the back and ear skin. In addi-
tion, AR extracts also caused an inhibition
of histological changes induced by repeated
application of D. farinae and a reduction
of total IgE and histamine levels in plasma
(P < 0.05). Furthermore, NO production
in LPS-treated RAW 264.7 cells was
diminished in a dose-dependent manner by
AR extract, and hTARC protein produc-
tion and TARC and MDC mRNA levels
in TNF-alpha/IFN-gamma-treated HaCaT
cells were also diminished by AR. The
inhibitory effect of AR on NO production
and TARC and MDCproduction may be
associated with the suppression of AD-like
skin lesions in D. farinae-induced NC/Nga
mice.
Conclusion: Topical AR was shown to be
effective in treating AD-like skin lesions in
NC/Nga mice and inhibited AD-related
gene (TARC and MDC) expression in
TNF-alpha/IFN-gamma-treated HaCaT
cells. Topical application of AR may there-
fore be a novel approach to the treatment
of AD.

Managing atopic dermatitis
1629
Taking probiotic yogurt by pregnant
mothers and their offspring for the first 6
months of life was associated with
reduced skin hypersensitivity to
histamine in the offspring
Shinohara, M1; Saito, H2; Matsumoto, K2
1 wheal size and the frequency of taking
Shimosizu National Hospital, Department of Allergy
and Pediatrics, Yotsukaido, Japan; 2National Research
Institute for Child Health and Development, Department
of Allergy and Immunology, Tokyo, Japan
Background: The intestinal microbiota is
known to play an important role in the
development of immune cells, including
dendritic cells and regulatory T cells. Previ-
ous intervention studies demonstrated that
oral administration of probiotics to the
pregnant mothers at risk during late preg-
nancy and mothers and/or offspring during
lactation period reduced the prevalence of
atopic dermatitis (AD) in the offspring,
whereas the incidence of atopy or the
development of other allergic diseases was
not altered. Results from these studies sug-
gest that administration of probiotics to
the pregnant mothers or the newborn
infants may prevent the development of
AD through mechanisms other than the
suppression of allergic sensitization. To
further clarify the mechanisms how probi-
otics affect the onset of AD, the associa-
tion between the intake of yogurt by
pregnant mothers and by the offspring,
and the skin reactivity to histamine in the
offspring was determined.
Methods: A total of 197 pairs of mothers
and their offspring (age 6.018.4 months)
were enrolled in this retrospective cohort
study. Infants with physician-diagnosed
AD and aged less than 6 months were
excluded. Mothers lled out a self-written
questionnaire about family history of aller-
gic diseases, and frequencies of taking
milk, fermented milk and yogurt during
late pregnancy by pregnant mothers and
the rst 6 months after the delivery by
mothers and/or the offspring. Wheal size
15 min after a skin prick test with 1 mg/ml
histamine and bifurcated needles was mea-
sured.
Results: The wheal size was signicantly
smaller in the offspring who or whose
mothers took yogurt everyday for the rst
6 months of life than in those who or
whose mothers did not (median, 2.60 mm
Allergy 66, Suppl. 94 (2011): 482642  2011 John Wiley & Sons A/S
Poster Session 71
versus 3.96 mm; P = 0.012). No signi-
cant differences were found between those
of offspring who or whose mothers took
milk or fermented milk and who or whose
mothers did not (median, 4.00 mm versus
4.00 mm; P = 0.329). A signicant nega-
tive correlation was found between the
yogurt in the offspring whose mothers had
taken yogurt few times during late preg-
nancy (P = 0.036).
Conclusions: Our results imply that taking
yogurt by the pregnant women and/or the
offspring may reduce the skin hypersensi-
tivity to histamine in the offspring and
thereby reduce the prevalence of AD.
1630
Nutrition economic impact of a specific
mixture of prebiotic oligosaccharides in
primary prevention of atopic dermatitis
Lenoir-Wijnkoop, I1; Nuijten, M2; Van Aalderen, W3;
Klaassen, D4; Sprikkelman, A3; Boehm, G5
1
Danone Research, Scientific Affairs, Palaiseau, France;
2
Ars Accessus Medica, Jisp, Netherlands; 3Emma
Childrens Hospital, Academic Medical Center, Pediatric
Respiratory Medicine and Allergy, Amsterdam,
Netherlands; 4Danone Research, Centre for Specialised
Nutrition, Wageningen, Netherlands; 5Danone
Research, Centre for Specialised Nutrition,
Friedrichsdorf, Germany
Background: In previous studies we have
shown clinical evidence that an infant for-
mula containing a specic mixture of pre-
biotic oligosaccharides (8 g/L scGOS/
lcFOS, ratio 9:1) is effective in the primary
prevention of atopic dermatitis, both at six
months and after 2 years. Now we aimed
to evaluate the cost-effectiveness of the use
of this specic infant formula for the pri-
mary prevention of atopic dermatitis in
The Netherlands.
Methods: We developed a model to esti-
mate the health economic impact of prebi-
otic preventive disease management of
atopic dermatitis. Data sources used
include published literature, clinical trials,
ofcial price/tariff lists and national popu-
lation statistics. The model takes into
account a cohort of children at risk for
allergy, in line with the population as men-
tioned in the clinical studies above. The
cost-effectiveness was calculated using a
Markov model reecting treatment pat-
terns and outcomes in the management of
atopic dermatitis. The analysis is con-
ducted from the perspective of the health
insurance in The Netherlands in 2009.
Results: The results show that the use of
an infant formula with 8 g/l scGOS/lcFOS
(9:1) leads to an increase in Quality
Adjusted Life Years (QALY) of 0.108,
when compared with the use of the same
infant formula without this prebiotic mix-
ture. Also, it results in an Incremental
Cost-Effectiveness Ratio (ICER) of 472.
The sensitivity analyses show that the
ICER remains in all cases far below the
Dutch threshold of 20,000/QALY.
Conclusion: This assessment shows that, in
addition to the previously reported health
benets and the improved quality of life
during childhood, the use of an infant for-
mula containing 8 g/L scGOS/lcFOS (9:1)
also results in favourable nutrition eco-
nomic outcomes in children at risk for
allergy. Furthermore the study demon-
strates that the use of this specic prebiotic
oligosaccharide mixture is a highly cost-
effective way of preventing atopic dermati-
tis in The Netherlands. On the long term
this may lead to substantial cost savings
for the health care budget.
1631
Skin sensitivity to acetylcholine in atopic
dermatitis and pollinosis patients
Grebenchenko, E1; Fedenko, E2; Gushin, I2
1
Institute of Immunology, Skin allergy department,
Moscow, Russian Federation; 2Russian Federation
Background: The aim of the study was to
estimate skin sensitivity to acetylcholine
(ACH) both in atopic dermatitis (AD) and
pollinosis (PLN) patients and the inuence
of sensitization on it.
Methods: Sixty AD patients with HDM
and pollen sensitization, ten PLN patients
and ten healthy persons were included in
the study. Microcirculation in two symmet-
ric sites of the intact skin area of forearms
was estimated with laser doppler owmetry
in standard conditions. Then 20 microliter
0.5 M of ACH solution was injected intra-
cutaneously into a back surface of the left
forearm (for control the same volume of
0.9% NaCl solution was injected into a
symmetric point of the right forearm) and
during 20 min the skin reaction was mea-
sured with microcirculation index (1 sm
proximately ACH injection) and local sen-
615

sation was estimated by means of visual
analog scale.
Results: Atopic dermatitis patients felt an
itch, PLN patients and healthy persons -
tingling and burning in the ACH injection
site. The duration of this reaction corre-
sponded as follows: AD patients > PLN
patients > healthy persons. Changes of
microcirculation index on ACH injection
in AD patients during relapses were more
apparent, then in remission, and in PLN
patients in remission were more apparent
then in healthy persons.
Conclusionss: Reaction on ACH in AD
patients (itch sensation) is distinct from
that in PLN patients and healthy persons
(tingling and burning sensation). This dif-
ference is not connected with a sensitiza-
tion and can explain the cholinergic nature
of itch - basic AD symptom.
1632
Influence of topical corticosteroids and
zinc pyrithione on the yeasts malassezia
spp growth in patients with atopic der-
matitis
Golysheva, E
Mechnikovs Research Institute for Vaccines and Sera,
Moscow, Russian Federation, Allergy Department,
Moscow, Russian Federation
It is well known the role of yeast of Mala-
ssezia spp as a triggering factor in atopic
dermatitis (AD). Topical steroids (TS) -
are the rst choice in AD management.
Aim: To evaluate the effect Mometasone
furoate (Mf) and Zinc pyrithione (Zp) on
the yeasts growth in vitro and skin yeasts
density colonization on the skin of the
patients with AD. Patients. There were 30
patients (pts) with AD observed. The treat-
ment was provided during 14 days twice
daily. The skin lesion on the left arm was
treated by unguent of Mf, on the right arm
- by unguent of Zp. The samples for myco-
logical investigation were collected from
the skin from both arms before the treat-
ment and in 14 day after treatment.
Methods: The samples for the mycological
investigation were assayed from the
affected skin sites in the arms. Specic
identication of the yeast of Malassezia
spp were provided according to morpho-
logical, physiological, and taxonomic and
PCR signs. The culturing of Malassezia
species was provided in three mediums:
selective, selective mixed with Mf, selective
mixed with Zp.
Results: There were identied M. sympodi-
alis most frequently (in 61%). There was
no difference between yeasts diversity in
different arms before treatment. The spe-
cies Malassezia did not inuence on the
severity of AD. The density of yeast on the
616
skin before treatment varied 104108
CFU/sm2 (Me: 106 CFU/sm2). After treat-
ment by Mf the density varied from 106 to
109 CFU/sm2 (Me: 10.8 CFU/sm2). After
treatment by Zp the density varied from
102 to 105 CFU/sm2 (Me: 10.4 CFU/sm2).
(P < 0.05). There was stimulating effect
on Malassezia growth in medium with Mf
in comparison with supressive effect in
medium with Zp in all plates.
Conclusions: TS activates the growth of
Malassezia spp. on the skin surface and
in vitro experiments. This effect may under-
lie in withdrawal syndrome after long-term
treatment with TS in patients with AD sen-
sitized to Malassezia. Contrary abscopal
effect of Zp on yeasts growth is useful in
treatment of patients with clinical relevant
yeasts sensitization.
1633
Fungal allergy and clinical severity in 523
patients with atopic dermatitis
Nahm, D; Hwang, E; Kim, J; Kim, J; Jin, H; Ye, Y; Park,
H gic constitution in children of an early age
Ajou University Hospital, Department of Allergy and
Rheumatology, Suwon, Republic of Korea
Background: Allergic responses to house
dust mite (HDM) and Malassezia have
been regarded as important aggravating
factors in adult patients with atopic derma-
titis (AD). However, a clinical signicance
of allergic response to fungus in AD is not
still established.
Methods: To evaluate a clinical signicance
of fungal allergy in AD serum levels of
specic IgE antibodies to Malassezia, Can-
dida, Trichophyton, Dermatophagoides pter-
onyssinus, and Dermatophagoides farinae
were measured by CAP-FEIA and clinical
severity of AD was assessed by the severity
scoring of atopic dermatitis system (SCO-
RAD) in 523 patients with AD. A level of
allergen-specic IgE antibody same or
above 3.5 kU/l was regarded as positive
result.
Results: In patients with AD, the positive
rate of serum allergen-specic IgE antibody
test was 65.0% for D. farinae, 63.5% for
D. pteronyssinus, 50.9% for Malassezia,
38.2% for Candida, and 23.1% for Tricho-
phyton. Level of specic IgE antibody to
fungal allergen was higher than level of
specic IgE antibody to HDM in 120
patients with AD (22.9%). SCORAD val-
ues signicantly correlated with levels of
specic IgE antibodies to Candida
(R = 0.365), Trichophyton (R = 0.353),
Malassezia (R = 0.304), D. pteronyssinus
(R = 0.244), and D. farinae (R = 0.231)
(P < 0.001). Multivariate binary logistic
regression analysis showed that a presence
of Candida allergy (positive specic IgE
antibody test) was the most signicant
Allergy 66, Suppl. 94 (2011): 482642  2011 John Wiley & Sons A/S
Poster Session 71 Managing atopic dermatitis
indicator distinguishing patients with
severe AD (SCORAD>50) from patients
with mild to moderate AD (adjusted odds
ratio, 2.2; 95% CI, 1.34-3.71; P = 0.002).
Conclusions: Fungal allergy (especially
Candida allergy) should be evaluated in
patients with severe AD. The clinical sig-
nicance of Candida allergy in atopic der-
matitis needs to be further studied.
1634
Features of specific examination and
treatment of patients with atopic derma-
titis associated with bronchial asthma
Mavlyanova, S1; Razikova, I2
1
Center of Dermatology & Venereology, Skin Diseases
Departmant, Tashkent, Uzbekistan; 2Tashkent Medical
Academy, Asthma & Allergy Department, Tashkent,
Uzbekistan
Background: How it is known atopic der-
matitis is an integral part of atopic disease
combining different forms of chronically
lasting disease with specic allergic reac-
tions. Atopic dermatitis arising with aller-
is with the course of time transforming and
may acquire such different forms as rhinitis
and asthma. Searching for new methods of
treatment of atopic dermatitis remains
actual because of atopic dermatitis is aller-
gic march for development of such terrible
disease as bronchial asthma.
Methods: Scientic investigations of very
many researchers show that an early spe-
cic diagnosis and treatment with allergens
leads to reduction of microorganisms sen-
sibilization spectrum, stops transition of
atopy into asthma and, naturally, to per-
fection of quality of life of patient. Specic
examination (collection of allergic anamne-
sis) was performed in specic group of
patients with diagnosis atopic dermatitis +
atopic bronchial asthma (36 patients) aged
from 5 to 20. They were distributed as fol-
lows: children up to 7 years (10), from 7 to
10 (9) and over 10 years (17 subjects).All
the patients depending on therapy obtained
were divided into 2 groups. The 1st group
of patients (17 subjects) received conven-
tional therapy (antihistamines, diet, adsor-
bents and external steroid preparations).
The 2nd group (19 subjects) received spe-
cic vaccination with allergens (SVA) tak-
ing into account sensibilization, i.e. these
patients received combined specic and
non-specic therapy.
Results: After therapy SCORAD was
decreasing at an average 68% in patients
obtaining combined treatment (SVA +
conventional), whereas only 32% in the 1st
group. The nearest results of treatment
exhibited clinical recovery in the 1st group
in six, sufcient improvement in seven
and improvement in three patients. In the
Poster Session 71 Managing atopic dermatitis
2nd group restoration was observed in 16,
demonstrable perfection - in three patients.
No adverse effects during and after com-
bined treatment were registered.
Conclusions: Therefore, our experience of
treatment and observation showed that
combined treatment of patients with AD
resulted in improvement of common status
due to striking reduction of skin manifesta-
tions and events of rhinitis and asthma.
1635
Improvement of atopic dermatitis sever-
ity after reducing indoor air pollutants in
nine kindergartens in Seoul, Korea
Park, C; Cho, S; Chung, B; Ahn, I; Kim, H; Lee, C
Kangnam Sacred Heart Hospital, Dermatology, Seoul,
Republic of Korea
Background: Genetic and environmental
factors contribute to the occurrence of ato-
pic dermatitis (AD). Recent epidemiologic
studies have shown that environmental
contaminants such as air pollution and
tobacco smoke play an important role in
the pathophysiology of AD.
Methods: The study population consisted
of 425 children in nine kindergartens,
Guro-gu, Seoul, Korea. The authors sur-
veyed the prevalence of AD and evaluated
disease severity by the EASI score and
investigators global assessment (IGA).
Indoor air pollution was measured and the
program to improve indoor air quality was
conducted in nine kindergartens. Seven
months later, the prevalence and disease
severity were evaluated.
Results: The initial prevalence of AD was
8% and the mean EASI score was 2.37.
The levels of particulate material ten
(PM10) and carbon dioxide (CO2) were
higher in some kindergartens than normal
values. After the program for improving
indoor air quality, the mean PM10 level
was signicantly decreased from 182.7 to
73.4 lg/m3. However, the changes in CO2
levels were variable. After the program, the
prevalence of AD was decreased to 7.6%
and the mean EASI score was decreased to
1.19. During the rst survey, there were 20
IGA 1 (almost clear) AD patients, eight of
IGA 2 (mild) AD patients and six IGA 3
(moderate) AD patients, but during the
second survey, the patient distribution
changed to nine IGA 0, (clear) AD
patients, 15 IGA 1 AD patients, 5 IGA
two AD patients and 2 IGA three AD
patients. The mean number of hospital vis-
its decreased from 1.3 per month during
the rst survey to 0.7 per month during
the second survey, which was statistically
signicant.
Conclusions: Indoor air pollution could be
related to AD. Reduction of PM10 by
Allergy 66, Suppl. 94 (2011): 482642  2011 John Wiley & Sons A/S
improving indoor air quality should be looks more promising, namely in case of
considered in kindergartens and schools in allergy to house dust mites.
order to prevent and relieve AD in chil- Conclusion: Our results are only prelimin-
dren. ary; to obtain more powerful data, it is
necessary to continue in the study and
enlarge the study group. However, observ-
ing SCORAD changes depending on aller-
1636
gen exposure could be a good tool for
The diagnosis of hypersensitivity in chil-
dren with atopic dermatitis and assess-
allergen clinical relevance assessment, espe-
ment of its relevance cially in house dust mites. Conversely, the
changes in need for anti-inammatory
Liska, M1; Gutova, V1; Panznerova, P2; Panzner, P1
1 treatment do not seem to be suitable for
University Hospital Pilsen, Department of Immunology
and Allergology, Pilsen, Czech Republic; 2University this purpose.
Hospital Pilsen, Department of Dermatovenereology,
Pilsen, Czech Republic
Background: Atopy patch test (APT), skin 1637
prick test (SPT) and specic IgE measure- Decline in Staphylococcus aureus
ment are used for examination of hyper- susceptibility to antibiotics in children
with atopic dermatitis
sensitivity in atopic disorders such as
atopic dermatitis (AD). However, it is Treneva, M1; Pampura, A1; Zaporozhtseva, Z2
1
Moscow Scientific Reasearch Institute of Pediatrics
often difcult to assess the clinical rele-
and Child Surgery, Allergy and Clinical Immunology
vance of the results. We started a study of Department, Moscow, Russian Federation; 2Moscow
AD children to investigate the clinical rele- Scientific Research Institute of Pediatrics and Child
vance of hypersensitivity tests. Surgery, Laboratory Department, Moscow, Russian
Federation
Methods: A group of 11 children with AD
was tested for allergy to four common ae-
Background: Staphylococcus aureus (S. aur-
roallergens (grass and birch pollen,
eus) colonization is revealed in 8790%
Der.pteronyssinus and Der.farinae) using
patients with atopic dermatitis (AD).
APT, SPT and specic IgE measurement.
S. aureus antibiotic susceptibility proles
A history of AD worsening after allergen
are different and could not be generalized
exposure was obtained using standardized
to majority of clinical settings or regions.
questionnaire. Additionally, clinical score
Investigations in dynamics of S. aureus sus-
of AD (SCORAD) and the number of days
ceptibility to antibiotics and time limitation
with need for topical anti-inammatory
of this process is of value in children with
treatment (AITD) were recorded during
AD.
1 year. The period covered both pollen sea-
Methods: For 20072009 years 646 skin
son and 6 weeks of mite avoidance regi-
cultures of children with AD were exam-
men. The effectiveness of the regimen was
ined retrospectively. In 248 cultures
conrmed by Der p 1 and Der f 1 concen-
S. aureus susceptibility to antibiotic was
trations assessment. We aim to nd out the
performed. Repeated within 0.530 months
specicity and sensitivity of APT, SPT and
30 cultures of S. aureus with it susceptibil-
specic IgE based on history, changes of
ity to antibiotic were obtained in 22 chil-
SCORAD and AITD; however, so far it is
dren. So 52 resistance patterns (22 initial
not calculated due to a low number of
and 30 repeated within 0.530 months time
enrolled subjects.
interval) to 19 antibiotics were analyzed.
Results: Four patients completed the whole
S. aureus antibiotic resistance versus
observational period and seven patients
months passed since primary test was stud-
completed 6 weeks of mite avoidance regi-
ied with correlations (Spearman, Kendall
men. APT was positive in nine patients
tau, gamma).
and the most common positive allergens
Results: Negative correlation of S. aureus
were house dust mites (n = 7). At least
antibiotic susceptible strains prevalence
one test of immediate-type hypersensitivity
versus months passed since initial pattern
(SPT, specic IgE) was positive in ve
is revealed (c = )0.24, P < 0.037; s =
patients, the most frequently to grass pol-
)0.199,P < 0.037). Initial prevalence of
len (n = 4). Mite concentrations decreased
antibiotic susceptible strains was prevailed
signicantly after avoidance regimen imple-
over resistant ones. 12 months later this
mentation; stronger impact on SCORAD
difference became not reliable. Increase of
values was apparent in patients positive to
S. aureus resistance against ampicillin and
house dust mites both in APT and SPT/
susceptibility to ciprooxacin were reliable
specic IgE. The assessment of clinical rel-
(P < 0.043 and P < 0.02).
evance on the basis of AITD seems to be
Conclusions: A child with AD is needed to
improper since the differences of AITD
be set and clear up with anti-inammatory
between patients were not signicant. The
and antibacterial therapy in a 12 month
assessment based on SCORAD changes
period past hospital check-up. The
617

decrease of S. aureus antibiotic susceptible
strains within this period could probably
reduce the effect of the following antibacte-
rial therapy.
1638
Staphylococcus aureus susceptibility to
antibiotics in children with atopic
dermatitis comparison between 200204
and 200709 years
Treneva, M1; Pampura, A1; Fedenko, E2; Voronina, V3;
Elyasova, M4
1
Moscow Scientific Research Institute of Pediatrics and
Child Surgery, Allergy and Clinical Immunology
Department, Moscow, Russian Federation; 2Scientific
State Institute of Immunology, Moscow, Russian
Federation; 3Moscow Scientific Research Institute of
Pediatrics and Child Surgery, Dermatology Unit,
Moscow, Russian Federation; 4Moscow Scientific
Research Institute of Pediatrics and Child Surgery,
Laboratory Department, Moscow, Russian Federation
Background: Staphylococus aureus (S. aur-
eus) colonization is revealed in 8084% of
patients with atopic dermatitis (AD). There
are no research results on S. aureus suscep-
tibility to antibiotics with a several years
cover period. A research of S. aureus sus-
ceptibility to antibiotics within the period
from 200204 years to 200709 years is of
value in children with AD.
Methods: S. aureus susceptibility to antibi-
otics was evaluated in 62 children with AD
in 200204 years and in 199 children in
200709 years. Correlation methods by
Spearman, Kendall tau and gamma-corre-
lation were used to compare S. aureus sus-
ceptibility 200204 versus 200709 years.
Results: Positive correlations of S. aureus
antibiotic susceptibility 200204 versus
200709 was revealed for penicillin (0.75
0.91 P < 1 1017), ampicillin (0.610.76
P < 1 1017), erythromycin (0.530.91
P < 1 1017), linkomycin (0.270.57 P <
0.0003) and laevomycethin (0.710.98 P <
1 1017). There was no correlation for oxa-
cillin and cefuroxime. Negative correlation
was revealed for ceftriaxone ()0.11 and
)0.28 P < 0.01) and ciprooxacin ()0.43
and -,76 P < 1 1017).
Conclusions: Staphylococcus.aureus antibi-
otic susceptibility in 20072009 versus
200204 is reduced to penicillin, ampicillin,
erythromycin, linkomycin and laevomyce-
thin in AD children. Susceptibility to oxa-
cillin and cefuroxime is not changed.
S. aureus susceptibility to ceftriaxone and
ciprooxacin increased in 20072009 versus
200204. Ceftriaxone should be recom-
mended as antibiotic of choice in children
with infected AD. Oxacillin and ce-
furoxime should be recommended as an
alternative choice.
618
1639
Anti-IgE (omalizumab) effective as atopic
dermatitis treatment: A case series
Rodrguez Rodrguez, M; Antoln Amerigo, D;
Barbarroja, J; Mohedano Vicente, E; Perz Bustamante,
M M2; Gonzalez Gutierrez, M1; Vazquez Cortes, S3
Prncipe de Asturias University Hospital, Allergy (ESI-
A), Alcala de Henares. Madrid, Spain
Background: Atopic dermatitis (AD) is a
chronic cutaneous multifactorial inamma-
tory disease characterized by pruritic skin
lesions frequently associated with food or
respiratory allergy which usually elicits
unbearable morbidity. Due to similarities
in the immune patterns of asthma and AD,
anti-IgE (omalizumab) may be effective in
the latter. We ought to evaluate Oma-
lizumab efcacy on Severe Atopic Derma-
titis patients who suffer from allergic
disease by means of quality of life ques-
tionnaires and physician overall assessment
Methods: We present a series of four
patients suffering from recalcitrant and
extremely severe Atopic Dermatitis associ-
ated with atopic diathesis related to peren-
nial allergens which do not respond to
diverse treatment options. Age range was
2542 years. Total IgE ranged from 472 to
12900 KU/l. All four patients were receiv-
ing either systemic or topical corticoster-
oids associated with antihistamines and
were previously treated with UVB radia-
tion or oral Cyclosporine showing absence
of improvement. Anti-IgE (Omalizumab)
300 mg was administered every other week
or once a month. Dermatology Life Qual-
ity Index (DLQI) and Quality of Life
Index for Atopic Dermatitis (QoLIAD)
were assessed at rst visit and 14 weeks
after treatment induction, in order to bet-
ter appraise and direct therapy response.
Results: Physician overall assessment
improved in 100% of patients. Dermatol-
ogy Life Quality Index (DLQI) scores were
two-fold lower 14 weeks after treatment
induction in three of them. The Quality of
Life Index for Atopic Dermatitis (QoL-
IAD) depicted substantial reduction when
measured before and after omalizumab
introduction (one to ve point-decrease).
All four patients displayed objective ame-
lioration of Atopic Dermatitis signs even
after corticosteroids interruption and occa-
sionally reported antihistamines demand.
Conclusions: We present a series of recalci-
trant Atopic Dermatitis associated with
atopic diathesis due to perennial allergens
in which Omalizumab became essential so
as to manage them satisfactorily. Anti-IgE
treatment may be a safer option to conven-
tional unresponsive therapies. Prospective
follow-up in these patients should be per-
formed in order to withdraw nal out-
comes.
Allergy 66, Suppl. 94 (2011): 482642  2011 John Wiley & Sons A/S
Poster Session 71 Managing atopic dermatitis
1640
Recalcitrant atopic dermatitis treated
with omalizumab
Moreira Jorge, A1; Robledo Echarren, T1; Vazquez
Cortes, S2; Gonzalez Gutierrez, M1; Martnez Cocera,
1
Hospital Clinico San Carlos, Allergy, Madrid, Spain;
2
Spain; 3Hospital del Henares, Allergy, Madrid, Spain
Background: Atopic dermatitis (AD) is a
chronic cutaneous inammatory disease.
Various categories of therapeutic medica-
tions are used for treating AD. Oma-
lizumab (Xolair, East Hanover, NJ;
Genentech, South San Francisco, CA) is a
monoclonal anti-IgE antibody that binds
to IgE molecules at the high-afnity recep-
tor (FcRI) binding site. Therefore, oma-
lizumab can be used as a potential new
systemic treatment agent for recalcitrant
AD with elevated IgE levels.
Methods: We report a 38-year-old man
who presented with chronic severe atopic
dermatitis. Failed treatments for this
patient included topical corticosteroids,
topical pimecrolimus, oral prednisone, oral
antihistamines, oral corticosteroids and
cyclosporine. Only intravenous corticoster-
oids provided signicant relief. The patient
had an immunoglobulin E (IgeE) level
higher than 25.000 IU/ml., and a severe
chronic dermatitis with an SCORAD index
of 47.29. We started treating him with
Omalizumab 450 mg subcutaneously every
2 weeks.
Results: Since the rst month of treatment,
he notably improved his symptoms, spe-
cially pruritus and insomnia. After 1 year
of treatment, the SCORAD index
decreased to 0. He is currently receiving
for 3 years, 450 mg every 4 weeks, and he
is still asymptomatic, with no skin lesions
and he didnt suffer of adverse effects.
Conclusions: We report the rst long term
(42 months) successful treatment of severe
AD with Omalizumab in Spain. We sug-
gest that omalizumab may have a role in
the treatment of isolated atopic dermatitis
in the adult population.
1641
The efficacy of wet wrap treatment in
children with atopic dermatitis
Song, T
Ilsan Paik Hospital, Inje University College of Medicine,
Division of Allergy and Respiratory
Medicine,Department of Pediatrics, Ilsan, Republic of
Korea
Background: Wet wrap treatment is a well-
known treatment for severe atopic dermati-
tis. But wet wraping with usual bandage
was troublesome and time-consuming pro-
cess of application. In this study, we used
a new garment for wet wrap treatment in
Poster Session 71 Managing atopic dermatitis
korean children with atopic dermatitis to
evaluate the efcacy and handiness.
Methods: We evaluated 22 children with
moderate-to-severe atopic dermatitis. They
were instructed to bath and to applicate
topical steroid and emollient. Then, wet
garment applied overnight. After 1
2 weeks, the SCORAD index, handiness
and side effects were evaluated.
Results: After 12 weeks, signicant
decrease of 37.6 points in mean SCORAD,
26.2 points in mean objective SCORAD,
and 11.4 points in mean subjective SCO-
RAD were observed (P = 0.0001). Four
patients refused wet wrap treatment due to
expensive price of new garment and had
treated with same methods except wet
wrap treatment. Children without wet
wrap treatment also had shown signicant
decrease of 16.5 points in mean SCORAD,
12.8 points in mean objective SCORAD,
and 3.7 points in mean subjective SCO-
RAD after 12 weeks. But decreasing
points of SCORAD was signicantly
greater in children with wet wrap treat-
ment. There were no signicant side effects
except chilling with one patient. All
patients wore new garment easily in 2 min.
Conclusions: Wet wrap treatment with new
garment was effective for decreasing both
objective and subjective symptoms in Kor-
ean children with moderate to severe ato-
pic dermatitis.
1642
Effectiveness of interferon-alpha in the
treatment of atopic dermatitis in children
Sizyakina, L; Penechko, E
Rostov State Medical University, Department of Clinical
Immunology and Allergology, Rostov-on-Don, Russian
Federation
Atopic dermatitis (AD) is the most com-
mon form of allergic skin disease, which is
based on violation of the ratio of Th1 and
Th2 - variants of immune response with a
predominance of the last one. Drugs which
stimulate Th1 lymphocyte subpopulations
may be used to correct this ratio.
The background of the study is to esti-
mate the effectiveness of substitution of
interferon in the treatment of patients with
AD.
The method used in total, 25 children
aged 6 to 17 AD patients who were divided
into two groups. Patients of the rst group
received standard therapy, and the children
of the group II in addition to basic therapy
received interferon-alpha: 1 million ME -
two times a day 10 days, then to 500
thousand ME- two times a day 10 days,
then to 250 thousand ME- two times a day
10 days.
Allergy 66, Suppl. 94 (2011): 482642  2011 John Wiley & Sons A/S
The results: The severity of AD was evalu-
ated by the index SCORAD (41.32
2.62 c.u. in the rst group and 43.23
2.52 c.u. in the II group). The clinical
observation of patients revealed a decrease
in intensity of pruritus, sleep improvement,
reducing inammation, reducing the degree
of lichenication and dryness in all patients
after treatment. However, in group II,
these changes were statistically signicant
25 2.5 c.u., the content of IgE practi-
cally did not change in patients of both
study groups. Comparative analysis of ef-
cacy of interferon-alfa in complex therapy
of AD revealed increased content of CD3
+ and CD4 + lymphocytes compared to
the rst group (CD3+ 73.3 3.06% and
53.8 1.3%; CD4+ )26 1 5% and
35 2.1%, respectively) decrease in
CD16+ cells (11.3 1.7% and 17
1.6%, respectively). It must be emphasized
that the children of group II showed signif-
icant increase in Th1 12.3 1.3% com-
pared with group I subjects 8.7 1.1%,
while the number of Th2 decreased slightly
(2.6 0, 9% and 2.9 0.8%, respec-
tively).
Conclusions: Thus, it was shown not only
high efciency of replacement therapy with
interferon-alpha, decreasing the level of
immunosuppression, but also a more pro-
nounced clinical effect in the treatment of
AD.
1643
The use of alternative medicine in
pediatric patients with atopic dermatitis
Uysal Soyer, O; Aksoy, I; Dallar, Y
The Ministry of Health of Turkey, Ankara Education and
Research Hospital, Pediatric Clinic, Ankara, Turkey
Background: Atopic dermatitis (AD) is one
of the most common chronic inammatory
skin diseases during childhood. Although
alternative treatments are frequently
referred for chronic diseases, limited data
are available about alternative medicine in
childhood AD. Our aim was to determine
the frequency, variety and risk factors for
alternative medicine in children with AD.
Methods: Patients aged between 6 months
and 18 years, diagnosed with AD with at
least 3 months duration were enrolled in
the study. A survey concerning questions
about demographic characteristics and use
of alternative medicines was administered
to parents of children with AD. Disease
related features were noted by the physi-
cians.
Results: Between FebruaryAugust 2010,
68 patients [60.3% male, 1.33 year (inter-
quartile range; 0.795 years) were included.
The patients admitted for medical care for
3 (24) times because of AD in the last
year. Patients SCORAD index was 20
(16.128.8) and 23 of them had atopy
(33.8%). Thirty (44.1%) families suggested
that AD affects quality of life. Alternative
treatments were used by 20 (29.4%)
patients and praying was the most com-
monly used alternative method (n = 15).
Other methods were; applications of wheat
spraying, scratching, mud bathing, juice of
hibiscus, coal powder, chicken stool, olive
oil, spring water, and drinking black grape
juice. The most common reason to use an
alternative method was unresponsiveness
to medical treatment (80%). According to
families, three of children who used alter-
native medicine had some benet and two
children had cutaneous infection after
application of chicken stool and juice of
hibiscus. There was no relation between
use of alternative medicines and gender,
age, family history of AD, atopy, quality
of life, parental socio-economic level, SCO-
RAD index, and control status according
to physician and serum IgE levels.
Conclusions: Prolonged treatment duration
and delay in response to treatment in
chronic allergic diseases might compel par-
ents to use alternative methods in AD.
Interestingly, utilization of alternative med-
icines is not related with the severity of dis-
ease, socio-economic level and educational
background of the parents.
1644
Atopic eczema management: what do
patients value in eczema education
workshops?
Hudson, P1; Grillo, M2
1
Flinders Medical Centre, Allergy, Adelaide, Australia;
2
Flinders Medical Centre, Dermatology, Adelaide,
Australia
Background: Atopic Eczema (AE) is a
chronic relapsing disease with a high bur-
den of care. Treatment regimes are com-
plex and frequently associated with poor
patient compliance. In 2005, a nurse led
Eczema Education Workshop (EEW) was
evaluated through a randomized controlled
study of 61 paediatric patients. Eczema
severity (measured by SCORAD) was
shown to be reduced in patients who
attended the EEW. Since 2006 government
funding has supported a monthly, commu-
nity based, EEW as part of routine eczema
management. Aim: To assess the impact of
the EEW on participant-perceived knowl-
edge of eczema, causes and treatments.
Methods: All participants were invited to
complete an evaluation survey relating to
EEW content and method of delivery. The
EEW is a 2 h, face to face education pro-
gram for patients and carers of children
diagnosed with eczema. The EEW is facili-
tated by Dermatology and Allergy special-
619

ist nurses who provide evidenced based
information in an informal, interactive
environment. The EEW focuses on the
physiology of eczema, importance of the
skin barrier effect on eczema severity. Skin
care regimes are explored and an individ-
ual eczema management plan is developed.
In addition, information is provided on
trigger factors, topical steroids, infection,
wet wraps and complimentary treatments.
Treatment methods are demonstrated and
participants practise these under nursing
guidance. Patients are able to self refer to
the EEW which is provided free of charge.
Child minding facilities are provided by an
accredited volunteer service. The EEW is
also available on-line for patients/carers
who are unable to attend a face- to- face
session.
Results: Since 2006, an average of 40 par-
ticipants per year, have attended the EEW.
61 participants, primarily with children
aged 05 years completed the survey. They
reported the most value from information
relating to Skincare 76%, Topical Steroids
72%, Wet Wraps 64%, Infections 61%,
and Allergic Trigger factors 54%. These
participants believed they had a greater
overall understanding of eczema (2.84.7
out of a score of ve) after attending the
EEW.
Conclusions: The EEWs make a valued
contribution to improving carers knowl-
edge of eczema, causes and treatments and
potentially empower families to develop,
self management skills.
1645
Atopic dermatitis under systemic treat-
ment with tacrolimus - clinical case
Ghiordanescu, I; Bumbacea, R; Antonie, A
Elias Emergency Hospital, Dermatology and
Allergology, Bucharest, Romania
Background: Tacrolimus is a macrolide
agent now used as a primary immunosup-
pressant in prevention of graft rejection in
transplant recipients. It also complements
existing treatment options for atopic der-
620
matitis and has been accepted as a rst-line
treatment for inammation.
Methods: Clinical case: We present the
case of a 6-year-old male patient, with Ala-
gille syndrome (autosomal dominant disor-
der associated with abnormalities of the
liver, heart, kidney, and other systems) that
underwent hepatic transplantation (Decem-
ber 2005) which required long-term immu-
nosuppression with tacrolimus. The patient
has a history suggestive of atopic dermati-
tis (Hanin and Rajka Diagnostic Crite-
ria), with recurrent symptoms since 2008
and no family history of allergic diseases.
Laboratory nding showed elevated levels
of total IgE and eosinophilia (absent
before transplantation). Tacrolimus blood
level ranged 44.2 ng/ml. Skin prick-test
was not performed. The treatment con-
sisted of dermatocorticosteroids, emollients
and H1-antihistamines; parents refused
proactive therapy with tacrolimus. The
patients clinical evolution under treatment
was favorable but pruritus persisted.
Results and Discussions: Cyclosporine A
and tacrolimus, both calcineurin inhibitors,
have been used extensively for immunosup-
pressive therapy in pediatric liver trans-
plant recipients. They share a similar
mechanism of action, the inhibition of
cytokine gene transcription primarily inter-
leukin-2 in T lymphocytes. Despite the
strong immunosuppressive property, there
are several reports of food allergy, asthma
and eosinophilic colitis in pediatric trans-
plant recipients under Tacrolimus immuno-
suppression, but not Cyclosporine A.
Selective suppression of Th1 lymphocytes
by the IL-2 inhibitor immunosuppressant
tacrolimus could promote Th2 lymphocytes
and an allergic immune response. Eosino-
philia is present in up to 50% of children
and adolescents receiving tacrolimus immu-
nosuppression. The majority of these
patients also have elevated levels of total
and specic (mainly to food allergens) IgE
antibodies. Most patients are asymptom-
atic and do not manifest food allergy or
asthma.
Allergy 66, Suppl. 94 (2011): 482642  2011 John Wiley & Sons A/S
Poster Session 71 Managing atopic dermatitis
Conclusions: More studies are needed in a
controlled setting to identify the prevalence
of similar ndings among other pediatric
liver transplant recipients. Atopic dermati-
tis has a Th2 immune-reactivity pattern.
Tacrolimus, by promoting Th2 lympho-
cytes, could cause allergic immune
responses. Is there a link between the two
of them?
1646
Prevalence of seborrhoeic dermatitis in
hospitalised psychiatric patients and
comparison with normal population
Pezeshkpoor, F; Kiafar, B; Saadatian, V; Ebrahimzadeh,
M
Mashhad Ghaem Hospital, Mashhad, Islamic Republic
of Iran
Background: Seborrheic dermatitis is a
common chronic papulosquamous derma-
tosis that is usually easily recognized by
Pink lesions covered with yellow-brown
scales and crusts on sebaceous areas of the
body. The disease affects 35% of the pop-
ulation and is reported more commonly
with a series of under lying: HIV infection,
Parkinsonism, epilepsy, and a few other
neurologic abnormalities. The object of
this study is to determine the prevalence of
seborrheic dermatitis in hospitalized
Psychiatric patients and compare it with
normal population.
Methods: Fifty hospitalized Psychiatric
patients examined for signs of dermato-
logic disorders and compared to 100 nor-
mal persons.
Results: In Psychiatric patients group prev-
alence of seborrheic dermatitis was 2.5%,
with no statically signicant difference with
control group. Also, there was no differ-
ence in sex, and age of prevalence of se-
borrheic dermatitis in the patients and
control.
Conclusions: Seborrheic dermatitis was the
most common skin disease in psychiatric
patients, followed by pityriasis and contact
dermatitis.

New data on efficacy, safety and mechanistic aspects of
1647
The specific immunotherapy efficiency
assessment methods at patients suffer-
ing from pollinosis
Dytyatkovska, Y
Dnipropetrovsk City Clinical Hospital #7, City Allergy
Centre, Dnipropetrovsk, Ukraine
The aim of present study was dening the
immunologic and clinical criteria for SIT
(Specic Immunotherapy) efciency assess-
ment at patients suffering from pollinosis.
There were 96 patients under our observa-
tion aged from 18 to 50 years old. They
suffered pollinosis with sensitization to
ambrosia, sage-brush, orach, cyclachaena,
sunower and maize. All the patients had
been being observed during 5 years and
had been divided into 4 groups: 1-st group
- 43 patients who received one pre-seasonal
SIT course; 2-nd group - 16 patients who
received two pre-seasonal SIT courses; 3-rd
group - 12 patients who received 3 pre-sea-
sonal SIT courses; 4-th group - 19 patients
who received ve pre-seasonal SIT courses.
Before the 1-st and after each of the fol-
lowing SIT courses the indices of IL-4, IL-
5, IL-10, IL-12, IFN-c (interleukines 4, )5,
)10, 12 and interferon-gamma) in blood
serum had been studied. For QL (Quality
of Life) assessment the retrospective
patients inquiry had been held using the
quality-of-life questionnaires.
At all the patients the indices studied
improved. The IL-4 level reliably decreased
after the 2-nd SIT course (from
0.63 0.07 pg/ml to 0.46 0.01 pg/ml,
P < 0.049); IL-5 decreased reliably after
the 3-d SIT course (from 4.58 0.28 pg/
ml to 2.93 0.19 pg/ml, P < 0.02) - the
data above conrms the SIT impact on the
TH-2 (T-helper-2) activity decrease. IL-12
Table 1. For abstract 1647.
SIT course 1-st SIT course
QL assessment n = 43
Excellent 5 (11.6%)
Good 34 (79.1%)
Satisfactory 4 (9.3%)
Allergy 66, Suppl. 94 (2011): 482642  2011 John Wiley & Sons A/S
Poster Session 72
immunotherapy
increased reliably and signicantly after the
3-d SIT course (from 74.46 11.93 pg/ml
to 194.09 38.51 pg/ml, P < 0.012)
which points onto the TH-1(T-helper-1)
immune response way switch-on. IFN-c
had increased most signicantly after the
5th SIT course (from 3.35 0.37 pg/ml
to 10.67 1.19 pg/ml, P < 0.001) which
evidences the immune response switch
from the TH-2 to TH-1 way. All the
patients pointed at the QL improvement
after the 1-st, the 2-d and the 5-th courses.
The indices of IL-4, IL-5, IL-10, IL-12
and IFN-c dynamics can be immunologic
criteria for SIT efciency assessment at
patients suffering from pollinosis as well as
predictors of SIT courses quantity needed
to obtain a sufcient immunologic effect.
SIT with pollen allergens provides a suf-
cient QL improvement after the 1-st, the 2-
d and the 5-th courses.
1648
Changes in bronchial reactivity to
histamine in the course of allergen
immunotherapy in seasonal allergic
rhinitis patients - are they really caused
by the treatment? Comparison of two
schedules, maintenance versus
pre-seasonal
Swiebocka, E1; Siergiejko, P2; Rapiejko, P3;
Siergiejko, Z4
1
University Children Hospital, Pediatrics,
Gastroenterology and Allergology Department,
Bialystok, Poland; 2Bialystok University Hospital,
Internal Medicine and Rheumatology Department,
Bialystok, Poland; 3Military Institute of Medicine, ORL
Department, Warsaw, Poland; 4Medical University of
Bialystok, Respiratory System Diagnostic and
Bronchoscopy Department, Bialystok, Poland
Background: Specic subcutaneous allergen
immunotherapy (SCIT) is the therapeutic
2-d SIT course 3-d SIT course 5-th SIT course
n = 16 n = 12 n = 19
5 (31.3%) 1 (8.3%) 5 (26.3%)
8 (50.0%) 7 (58.3%) 11 (57.9%)
3 (18.8%) 4 (33.3%) 3 (15.8%)
method recommended in patients suffering
from seasonal allergic rhinitis (SAR) at
whose pharmacological treatment does not
bring satisfactory results. Usually injections
of allergen or allergoid extracts are given
in two schedules: pre-seasonal only or after
the build-up period, in monthly intervals
of the highest well tolerated dose of the
vaccine by the period of 3 to 5 years
(maintenance treatment). This study was
aimed to compare effects of the two sched-
ules of SCIT, on bronchial response to his-
tamine.
Methods: Fifty seven patients were
included into the study. They were
randomly divided into 2 groups: A (n =
29)-maintenance group and B (n = 28) -
pre-seasonal group. All of them were sensi-
tive to grass pollen, what was conrmed by
skin prick tests. Studies were started in
autumn of 2005 (in 41 patients) or 2006 (in
16 patients). Before starting of SCIT, out-
side pollination period (NovemberJanu-
ary), bronchial provocation tests with
histamine (according to Rosenthals
method) were performed at each patient.
Successive histamine challenges were
repeated at the same time of the year.
Every day the number of the grains of
pollen of grasses in one cubic metre of the
air was estimated in each season of polli-
nating.
Results: After 1st year of SCIT no signi-
cant changes in BHR were observed. After
the 2nd year non signicant increases in
BHR in both groups (more intensive in B)
were seen. After the 3rd season of SCIT a
decrease in BHR was observed in both
groups. In group A decrease in BHR was
signicant in comparison to baseline and
to the 2nd season. In group B signicant
decrease in BHR was noted only in com-
parison to the 2nd season. What was the
cause of the increase in BHR after the 2nd
season? Maybe it was the result of the
intensive pollination during 2007 season?
The following maximal numbers of grass
pollen granules in 1 cubic metre of the air
in successive years were found: in 2005
385, 2006198, 2007480, 2008248, and in
2009168. After separate analysis of results
in patients started SCIT in 2005 and in
2006 we found that in both groups the
621

most intensive BHR was noted after the
2nd year of SCIT.
Conclusions: The BHR to histamine after
3 years of SCIT was signicantly lower
than at starting only in the maintenance
group. The intensity of pollination in sum-
mer preceding the histamine challenge can
have some effect on bronchial reactivity.
1649
The values of total and specific immuno-
globulin E and immunoglobulin G4 in the
initial phases of various forms of subcu-
taneus immunotherapy
Bystron, J1; Peprnikova, J2; Pazderova, D2;
Rozehnalova, K2; Szotkovska, J3; Hermanova, Z3
1
University Hospital, Olomouc, Czech Republic;
2 1651
University Hospital, Allergology and Clinical
Immunology, Olomouc, Czech Republic; 3University
Hospital, Immunology, Olomouc, Czech Republic
Background: We monitored the production
of total IgE antibodies, specic IgE and
specic IgG4 antibodies. Our aim was to
determine the level and rate of production
of these antibodies under the inuence of
different forms of the initial phase of aller-
gen injection therapy.
Methods: In 50 patients with conrmed
hypersensitivity (SPT, specic IgE) to birch
pollen, grass, dust mites, and insect aller-
gen (bee or wasp), who started a specic
allergen immunotherapy in the autumn
winter 2009/2010, we monitored the pro-
duction of total IgE antibodies, specic
IgE and specic IgG4 antibodies. We com-
pared patients with classical initial phase
(reaching the maintenance dose after about
3 months), patients with initial phase of
cluster method (reaching the maintenance
dose after 1 month) and initial phase of
rush method (reaching the maintenance
dose in 1 day). Laboratory testing was per-
formed before the initial phase and then in
1 and 3 months after the initial launch
phase.
Results: During rush SCIT the values of
total IgE and specic IgE were rising
mostly within the rst month, mainly for
the AIT to birch pollen and dust mites,
nearly twice the level. Then continued to
rise even more, but more gradually. In
SCIT to insect allergen specic IgG4 values
have increased signicantly the rst month
(especially during rush and cluster meth-
ods) up to four times and after 3 months
up to ten times its original values. During
the classical initial phase there was only a
triple rise after 3 month of application. In
SCIT to tree and grass pollen there was a
steep increase in specic IgG4 only by rush
(after 3 months for more than ten-fold)
while the use of a standard procedure and
the use of the classical scheme or allergoid
standard course caused the 12 fold
622
Poster Session 72 New data on efcacy, safety and mechanistic aspects of immunotherapy
increase after 3 months of application. In
SCIT to dust mites after 3 months a signif-
icant difference between the rush form
(after 3 months a seven-fold sIgG4 rise)
and the classical form (only a slight sIgG4
rise) was registered.
Conclusions: The rapid initial phase of the
AIT applications (rush, cluster) induce a
much higher production of specic IgG4
than the classical initial phase. This fact
can be used mainly in cases where we want
to induce the protective effect of AIT as
soon as possible (e.g. allergy to insects or
pollen before the coming season).
1650
Amount of major allergen Bet v 1 and
allergen activity in different European
products for sublingual immunotherapy
Kerkvliet, E; Peekel, I; van Tuyn, J; Sleijster, H; van den
Hout, R
HAL Allergy BV, Development, Leiden, Netherlands
Background: Allergic rhinitis induced by
birch pollen can be treated by means of
allergen-specic immunotherapy. A num-
ber of products for sublingual immuno-
therapy (SLIT) are currently commercially
available. It has been acknowledged that
the dose for SLIT should be higher than
for subcutaneous immunotherapy and that
an adequate dose is decisive for clinical
efcacy. We investigated the content of the
major allergen of Betula verrucosa, Bet v 1,
and allergen activity in different SLIT
products.
Methods: SLIT products from main Euro-
pean allergy companies were purchased
and the concentration of Bet v 1 was deter-
mined using a validated ELISA method.
Bet v 1 is calculated in lg (microgram) per
maintenance dose, using the instructions
for administration as provided by the man-
ufacturers. The allergen activity was deter-
mined as stated in the Guideline on allergen
products1 using a validated method that is
based on a competition of IgE binding
between the SLIT products and a commer-
cial available biotin labelled allergen prepa-
ration. The IgE binding efciency (allergen
activity) is expressed by relative potency
(RP) of the product compared to a refer-
ence.
Table 1. For abstract 1650. Daily dose of Bet v1 and allergen activity per dose of 4
different Birch SLIT products
Product Strength
SLITonePLUS 2500 STU/ml
TOL SL plus 100 HEP/ml
Staloral 300 IR/ml
SUBLIVAC 10000 AUN/ml
Allergy 66, Suppl. 94 (2011): 482642  2011 John Wiley & Sons A/S
Results: The Bet v 1 amounts of Staloral
and SUBLIVAC represent a high dose of
major allergen. SUBLIVAC reveals the
highest Bet v 1 concentration per daily
dose. Allergen activity measurements of
the different products, also taking into
account the contribution of birch allergens
other than Bet v 1, are in agreement with
the Bet v 1 measurements.
Conclusions: We conclude that SUBLI-
VAC Birch has a higher strength per dose
than the other SLIT products. Guideline
on Allergen Products: Production and
Quality Issues, London, 20 November
2008, EMEA/CHMP/BWP/304831/2007
Abstract withdrawn
1652
Changes of skin reactivity and IgE level
during immunotherapy against home
dust and Dermatophagoides
pteronyssinus allergens
Prnjavorac, B1; Abduzaimovic, K2; Mujaric, E3; Causevic,
A4; Semiz, S4; Dujic, T4; Dzuvo-Hadzovic, A5; Jadric, R6
1
General Hospital Tesanj, Department of Internal
Medicine, Tesanj, Bosnia and Herzegovina; 2General
Hospital Tesanj, Transfusiology, Tesanj, Bosnia and
Herzegovina; 3Cantonal Hospital Zenica, Internal
Mediicine, Zenica, Bosnia and Herzegovina; 4Faculty of
Pharmacy, Biochemistry, Sarajevo, Bosnia and
Herzegovina; 5Faculty of Medicine, Physiology,
Sarajevo, Bosnia and Herzegovina; 6Faculty of
Medicine, Biochemistry, Sarajevo, Bosnia and
Herzegovina
Background: Immunotherapy against home
dust and dermatophygoides pteronissimus
could be performed separately or in mix-
ture. In twelve years experience of immu-
notherapy we observed increased reactivity
of skin after 1 year of treatment with
monthly injections, sometimes with need
for reduced doses of allergen for injections.
In the same time decreased level of IgE
was shown. But after 2 years of treatment
we observed decreased skin reactions in
these patients. How these changes in skin
reactivity were linked with IgE level is the
aim of this study.
Methods: The asthmatic patients with
known allergy to home dust and dermato-
phagoides pteronissimus allergy have been
Volume of Daily dose Daily dose allergen
the daily dose (ll) Bet v 1 (lg) activity (RP)
200 5.8 0.2
270 1.0 <0.1
400 25.4 0.6
200 46.7 1.0
Poster Session 72 New data on efcacy, safety and mechanistic aspects of immunotherapy
followed up for 5 years. After intracuta-
nous skin testing and measurement of
Immunoglobulin E (IgE) we performed
subcutaneous specic immunotherapy
(SCIT). Total and specic IgE were mea-
sured using enzyme-linked immunosorbent
assay (ELISA). First control of IgE was
performed after one year, second after
2 years, third after 3 years and next one
after 5 years of SCIT. Any time after SCIT
skin reaction was measured (diameter of
wheal) in millimeters (mm). Statistical
analysis was performed by ANOVA test of
analysis of variance for repeated measure-
ments.
Results: During period of 5 years we trea-
ted 74 patients with mixture of home dust
allergens. 41 were women and 33 men.
SCIT has been performed according to rec-
ommendation of EAACI for long course
of immunotherapy. Average baseline IgE
level were 526 IU/ml (SD 112), after 1 year
446 IU/ml (SD 94), after 2 years 392 IU/
ml (SD 85), after 3 years 367 IU/ml (SD
76), and after 5 years 266 IU/ml (SD 66).
Average diameter of the wheal were
13.15 mm (SD 2.1) after 1 year of immu-
notherapy 13.38 mm (SD 2.2), after 2;
after 3 years 12.7 mm (SD 1.8), and after 5
12.4 mm (SD 1.6). Not all patients deal the
same patterns of changes, but 14 out of 74
patients showed increased skin reaction
after 1 year. Statistical analysis using
ANOVA test showed statistical signicant
decrease (P < 0.05) of IgE level, but not
statistical signicant correlation between
IgE level and changes of skin reactivity.
Conclusions: During long term immuno-
therapy skin reactivity after SCIT could be
changed, with increased reactivity after
1 year of immunotherapy, but decreased
thereafter during next 4 years. Changes of
IgE levels did not in close correlation with
these changes of skin reactivity.
1653
The safety, tolerability and short-term
efficacy of subcutaneous mite immuno-
therapy in HDM-allergic patients
Kleine-Tebbe, J1; Nell, M2; van Ree, R3; Diamant, Z4;
Distler, A5; Boot, D2
1
Untersuchungszentrum Dermatologie, Allergologie
und Asthma (UZDAA), Berlin, Germany; 2HAL Allergy
BV, Medical Affairs, Leiden, Netherlands; 3Academic
Medical Center (AMC), University of Amsterdam,
Department of Experimental Immunology, Amsterdam,
Netherlands; 4Erasmus MC, Department of Allergology,
Rotterdam, Netherlands; 5HAL Allergie GmbH, Medical
Department, Dusseldorf, Germany
Background: The current maintenance dose
(10,000 AUeq/monthly) for house dust
mite (HDM) immunotherapy (IT) with the
subcutaneous allergoid PURETHAL
mites (PUR-M) has previously shown clini-
cal efcacy in patients with HDM-allergy.
Allergy 66, Suppl. 94 (2011): 482642  2011 John Wiley & Sons A/S
In order to comply with the EMA guide-
lines on IT products (2009), a study was
conducted to evaluate the safety and toler-
ability of increasing high PUR-M doses
(up to 40,000 AUeq).
Methods: Eligible symptomatic patients
(>18 years) with HDM-allergic rhinitis/
rhinoconjunctivitis with or without clini-
cally stable mild persistent asthma partici-
pated in this open-label study and were
randomized into one of three dosing regi-
mens: 1) a regular regimen with updosing
to 40,000 AUeq followed by two mainte-
nance doses (17 weeks), 2) an intermediate
regimen (14 weeks) or 3) a fast regimen
(11 weeks). Safety and tolerability were
evaluated by monitoring of early (swell-
ing> 5 cm, 15 min) and late (swelling
>5 cm, 24 h) local and systemic reac-
tions. In addition, short-term efcacy was
assessed by the Clinical Index Score (CIS)
and two surrogate efcacy markers: the
conjunctival provocation test (CPT) and
levels of serum allergen-specic IgG and
IgG4.
Results: In total, 39 (out of 48) patients
(25 males/14 females) completed the study
per protocol and received 440 subcutane-
ous injections of PUR-M. No early local
reactions occurred. Late reactions were
observed in 12% of the injections (7% in
the regular regimen, 11% in the intermedi-
ate regimen and 16% in the fast regimen).
In total, 31 systemic reactions, all grade 1,
were reported of which two needed oral
antihistamine treatment. No grade 2 sys-
temic reactions were observed. Six patients
did not reach the highest dose due to late
local reactions and/or systemic reactions
needing antihistamines (20% in the regular
regimen, 16% in the intermediate regimen
and 13% in the fast regimen). Higher
PUR-M doses were not associated with
substantially more drop-outs. Results of
CIS and CPT indicated efcacy of the
treatment at current dose and higher doses.
Serum immunoglobulin levels were
increased in all groups following treatment
(IgG4 Der p 25-fold and IgG4 Der f 21-
fold).
Conclusions: In an open-label study no
safety or tolerability issues were identied
for PUR-M doses up to 40,000 AUeq. No
differences in the safety aspects of the three
dosing regimens up to the maximum dose
were encountered. Therefore, this dose can
be safely included in future dose-range
nding studies.
1654
Changes in specific IgE to single grass
pollen allergens in patients undergoing
sublingual immunotherapy
Marcucci, F1; Sensi, L1; Incorvaia, C2; Soffia, S3; Scurati,
S3; Frati, F3; Di Cara, G1
1
Institute of Pediatrics, Department of Medical and
Surgical Specialties and Public Health, Perugia, Italy;
2
ICP Hospital, Allergy/Pulmonary Rehabilitation, Milan,
Italy; 3Stallergenes, Medical and Scientific Department,
Milan, Italy
Background: Allergen immunotherapy
induces a number of immunologic modi-
cations. Among these, allergen-specic IgE
antibodies generally show an increase dur-
ing the early phase of treatment, followed
by a subsequent progressive decrease. We
evaluated the changes in specic IgE (sIgE)
to single grass pollen allergens during the
rst year of treatment with sublingual
immunotherapy (SLIT) by a grass extract.
Methods: Thirty patients with grass pollen
allergy were included in the study. All
patients had persistent allergic rhinitis from
at least 2 consecutive years, graded moder-
ate-severe according to ARIA classica-
tion; 19 patients (14 males, ve females,
mean age 12.2 10.1 years) were submit-
ted to SLIT by a ve-grass pollen extract
(Staloral 300 IR, Stallergenes, Milan,
Italy). SIgE to single grass pollen allergens
(Phl p 1, Phl p 2, Phl p 5, Phl p 6, Phl p 7,
and Phl p 12) were measured by CAP Sys-
tem (Phadia, Uppsala, Sweden); 11 patients
(ve males, six females, mean age
10.5 3.9 years) served as controls. IgE
measurement was done at baseline and at
the end of grass pollen season.
Results: In patients treated with SLIT a
signicant increase in sIgE was observed
for Phl p 1 (P = 0.001), Phl p 2
(P = 0.001), Phl p 5 (P = 0.001), Phl p 6
(P < 0.001), while the IgEs for Phl p 12
and Phl p 7 were rarely present, always in
low amounts, and did not show new sensi-
tization after SLIT in negative patients. In
control patients no signicant increase in
sIgE for any single allergen was detected.
Conclusions: These preliminary ndings
conrm that the administration of an
immunologically effective allergen extract
by SLIT induces an early rise of sIgE and
show a pattern of IgE response to single
allergens characterized by signicant
increase to most, but not all, molecules.
623

1655
Tolerability of ultra-short updosing
phases in a compact sublingual immuno-
therapy dose regimen
Piller, M1; Preussner, L2; Drachenberg, K2
1
Bencard Allergie, Munich, Germany; 2Germany
Background: In sublingual immunotherapy
(SLIT) as well as in subcutaneous immuno-
therapy (SCIT) there are strong tendencies
to make dose regimens easier and more
convenient for patients in order to enhance
compliance. Moreover, with increasing use
of SLIT in recent years there is evidence of
a broader therapeutic range of the method
than formerly presumed. The tolerability
of two updosing regimens of a well-docu-
mented SLIT preparation in one day using
only the top concentration were investi-
gated.
Methods: Two hundred and thirty one new
SLIT patients, age 479 years, were treated
and interviewed in this prospective, multi-
centre, non-interventional trial. 53 centres
documented the updosing of SLIT for
patients with seasonal or perennial rhino-
conjunctivitis and/or asthma using one of
two regimens: a) 1-3-5-7 pumps and b) 1-3
pumps of top concentration, both with a
30 min observational phase between each
dose and after the last dose. All of these
treatment administrations were conducted
in the doctors surgery. After this initial
updosing the patients applied a) seven
pumps or b) three pumps daily at home.
The patients were questioned by the doctor
after 3 and after 12 weeks regarding tolera-
bility and adherence to the therapy regi-
men.
Results: 56% of the patients received the
1-3-5-7 regimen (Compact-Highdose) and
40% the 13 regimen (Compact). The
majority of doctors assessed the updosing
regimens as very good or good (85.7%); in
1.3% (3/231) of patients the updosing was
stopped. Adverse events were documented
for between 12% and 17% of single dos-
ing, predominantly mild local reactions
(tingling, itching, burning sensations).
There was one severe local reaction (itch-
ing) and only few mild systemic reactions.
In those patients who received the rst-day
therapy during their relevant pollen season
the adverse event rate was slightly higher
for the rst two doses but lower for the
doses three and four. The mean tolerability
of the at-home-therapy on a scale from
one (good)six (bad) was 1.4. Patients
acceptance of both therapies, with maxi-
mum doses of seven or three pumps, was
high (90.2%), although slightly better for
the Compact therapy (max 3 pumps).
624
Poster Session 72 New data on efcacy, safety and mechanistic aspects of immunotherapy
Conclusions: Ultra-short updosing regi-
mens with either four or two high-concen-
tration doses were well tolerated and
accepted, as well as the continuing daily
at-home therapy with either seven or three
pumps daily.
1656
Allergoids of Phleum pratense: investiga-
tions on their allergenic composition
Fernandez-Caldas, E; Tudela, J; Cases, B; Abel-
Fernandez, E; Casanovas, M; Subiza, J
Inmunotek S.L, R&D, Madrid, Spain
Background: The inhalation of grass pollen
is the main cause of allergic respiratory
diseases worldwide. Immunotherapy with
grass allergoids is widely used to treat
allergic rhinitis and allergic asthma. Allerg-
oids derive from natural proteins present
in Phleum pratense pollen grains, which are
polymerized with glutaraldehyde. The
molecular weight of these allergoids is
always greater than 100 kDa to avoid the
presence of non-polymerized allergens.
However, the presence of native allergens
in allergoid is still a matter of debate and
controversy. The aim of this study was to
determine the qualitative composition of
polymerized extracts using two purication
steps and Mass spectrometry.
Methods: The proteomic analysis was car-
ried out in the Proteomics Facility UCM-
PCM, Madrid, Spain. Briey, freeze dried
polymerized extracts were digested in tryp-
sin and the resulting peptides analyzed
using a nanoliquid chromatography-MS/
MS on an LTQ linear ion trap mass spec-
trometer. Peptides were separated on a
BioBasic C-18 PicoFrit column and
trapped and desalted. The mass spectrome-
ter was operated in the data-dependent
mode to automatically switch between full
MS and MS/MS acquisition. The parame-
ters for ion scanning were the following:
Full-scan MS (4001800 m/z) plus top
seven peaks Zoom/MS/MS (isolation width
2 m/z), normalized collision energy 35%.
The scanning was performed using a
dynamic exclusion list. MASCOT 2.1 was
used to search different databases with a
peptide tolerance of 300 ppm; fragment ion
tolerance, 0.6 Da; missed cleavage sites:
one, and xed modication, carbamidom-
ethyl cysteine and variable modications,
methionine oxidation. Protein scores are
derived from ions scores as a non-probabi-
listic basis for ranking protein hits. Indi-
vidual ions scores >5 indicate identity or
extensive homology (P < 0.05).
Results: The analysis of the peptides
revealed the presence of Phl p 1, Phl p 3,
Allergy 66, Suppl. 94 (2011): 482642  2011 John Wiley & Sons A/S
Phl p 5, Phl p 11 and Phl p 12. Other
potential allergenic proteins included: Eno-
lase, Fructose-bisphosphate aldolase,
Luminal-binding protein (Heat shock pro-
tein 70), Calmodulin, among others.
Conclusions: The MS/MS analysis conrms
the presence of native allergens in the poly-
merized extract of P. pratense.
1657
Sublingual specific immunotherapy in
intermittent and persistent allergic rhini-
tis: an observational case-control study
Mijakoski, D1; Karadzinska-Bislimovska, J2; Stoleski, S2;
Minov, J2; Marsenic, M1
1
Institute for Occupational Health of RM, WHO CC,
GA2LEN CC, Allergy Center, Skopje, Macedonia,
FYROM; 2Institute for Occupational Health of RM, WHO
CC, GA2LEN CC, Dpt for Cardiorespiratory Functional
Diagnostics, Skopje, Macedonia, FYROM
Background: Sublingual specic immuno-
therapy (SLIT) is a recognized and valid
treatment for allergic rhinitis and conjunc-
tivitis. Objective: to evaluate the efcacy of
SLIT in the reduction of symptoms and
rescue medications in patients with allergic
rhinitis (AR).
Methods: We performed a casecontrol
study and 30 patients with AR due to sea-
sonal or perennial allergens were analyzed.
Cases (n = 15) were dened as patients
with intermittent (60%) or persistent
(40%) AR treated for at least two consecu-
tive years with specic SLIT with the
related allergen extracts. Controls (n = 15)
were dened as sex, age, and type of aller-
gen matched AR patients who were never
treated with specic immunotherapy. Main
outcomes of the study were the rhinocon-
junctivitis symptom score (SS) (nasal con-
gestion, sneezing, rhinorrea, nasal itching,
ocular itching and watery eyes) with a
ranging scale from zero (no symptoms) to
three (severe symptoms) and the medica-
tion score (MS) describing symptomatic
drug intake (antihistamine and inhaled cor-
ticosteroids). SS and MS were evaluated at
the end of the observational period in rela-
tion with the period, considering the last
12 months, in which patients suffered the
highest symptoms levels (i.e., peak of rele-
vant pollen season in case of seasonal AR
or during the period of maximum allergen
exposure in case of perennial AR).
Results: The data obtained showed that
mean SS was 3.3 1.4 among cases and
6.7 1.8 in controls ()50.7%)
(P < 0.01). On the other hand, mean MS
was 1.9 1.5 and 3.1 1.8 in cases and
controls, respectively ()38.7%) (P < 0.05).
Conclusions: Our study showed that a 2-
year SLIT treatment in patients with
Poster Session 72 New data on efcacy, safety and mechanistic aspects of immunotherapy
intermittent or persistent AR is associated
with signicantly lower symptom and med-
ication scores in comparison with patients
treated with symptomatic drugs only.
1658
Epidemiological study on the use of
allergenic extracts of house dust mites
(Dermatophagoides farinae/
Dermatophagoides pteronyssinus)
adsorbed onto L-tyrosine in patients
suffering from allergic rhinoconjunctivitis
Alba, P1; Conejero, A2; Miras, M3; Caso, A4; Moreno, F5;
Dordal, T6; Zapata, J7; Amat, J8; Sanchez, G9; Orovitg,
A10; Iglesias, J11; Sanchez-Machin, I11; Gonzalez, R11;
Pozas, P11; Moreno, A12; Nunez, R13; Carballas, C13;
Anton, M14; Pamies, R15; Fraj, J16
1
Hospital de Manises, Manises (Valencia), Spain;
2
Clinica Alergoestudio, Sevilla, Spain; 3Sentro
Especialidades San Jeronimo, Sevilla, Spain; 4Sevilla,
Spain; 5Clnica Lobaton, Cadiz, Spain; 6Hospital
Municipal de Badalona, Badalona, Spain; 7Almera,
Spain; 8Hospital Torrecardenas, Almera, Spain;
9
Granada, Spain; 10CP Lux Sevilla, Sevilla, Spain;
11
Hospital La Candelaria, Santa Cruz de Tenerife, Spain;
12
Hospital de Cuenca, Cuenca, Spain; 13Hospital de
Caldes, Lugo, Spain; 14Hospital de Elda, Alicante,
Spain; 15H. La Lnea de la Concepcion, Cadiz, Spain;
16
Hospital Clnico Universitario Lozano Blesa, Zaragoza,
Spain
Background: Allergenic extract adsorbed
on L-tyrosine is a specic Immunotherapy
for the treatment of allergic rhinoconjunc-
tivitis and asthma. Available clinical stud-
ies demonstrate the efcacy of this
treatment but there are few data available
about the consultant and patient experi-
ence with the treatment applied on their
clinical practice, which was the main pur-
pose of this study.
Methods: An epidemiologic multicentre
study. 20 allergologists were asked to com-
plete a questionnaire that was separated
into two sections. The rst section
described the allergologists practice demo-
graphics, clinical practice and professional
experience and the second section
described patients demographics, treat-
ment type and patients experience with the
treatment. All patients had a diagnosed
allergy to Dermatophagoides farinae and/or
Dermatophagoides pteronyssinus.
Results: Nineteen allergologists completed
the questionnaires; all had at least 5 years
of experience and saw more than 15
patients suffering from rhinoconjunctivitis
per month. Those allergologists docu-
mented data from 113 patients. Allergolo-
gists used a cluster treatment schedule on
approximately half of the cases (54.6%;
n = 59), the others were on conventional
treatment schedule. According to the al-
lergologists, the main advantages of tyro-
Allergy 66, Suppl. 94 (2011): 482642  2011 John Wiley & Sons A/S
sine adsorbed vaccines when compared to
other subcutaneous vaccines were: good
tolerance (63.2%), free of aluminum
hydroxide (63.2%), exibility of dose
regimes (57.9%) and the fast updosing
phase (52.6%). Also allergologists stated
that the use of symptomatic drug therapy
was decreased by 50.7% after the adminis-
tration of allergenic extract adsorbed on L-
tyrosine. Regarding patient experience,
56.8% assessed the acceptance of treatment
as good and 34.23% as very good. Only
8.8% (n = 10) suffered from local reac-
tions and no patient suffered from a sys-
temic reaction.
Conclusions: The collected data can be
regarded as being representative of current
allergologists clinical practice. The aspects
described show that the initial treatment
with house dust mite allergenic extract
adsorbed onto L-tyrosine is perceived by
both the prescribing allergologist and the
patient receiving the treatment as a well
accepted therapeutic tool due to its advan-
tages and patient tolerability.
1659
Sublingual immunotherapy for respira-
tory allergies: a multicentre retrospective
safety survey
1 2 3
Torrecillas, M ; Martinez-Canavate, A ; Quiralte, J ;
Sarralle, R4; Manrique, I5; Justicia, J6; Garcia, M7
1
Hospital Perpetuo Socorro, Allergy Service, Albacete,
Spain; 2Hospital Materno Infantil Virgen de las Nieves,
Pediatric Allergic Service, Granada, Spain; 3Hospital
Virgen del Rocio, Allergy Service, Sevilla, Spain;
4
Private Clinic, Santander,Spain; 5Instituto Valenciano
de Pediatria, Valencia, Spain; 6Stallergenes Iberica,
Medical Department, Barcelona, Spain; 7Stallergenes
Iberica, Medical Department, Madrid, Spain
Background: Most data concerning adverse
reactions registered for sublingual allergen
immunotherapy (SLIT) have been
extracted from clinical trials and also from
spontaneous pharmacovigilance notica-
tions. The rst ones usually recruit not
enough patients to reect unfrequent reac-
tions and, in the second case, the number
of notications is, usually, less than
expected. This study pretends to bring the
gap between them collecting adverse reac-
tions in real clinical practice.
Methods: Post-authorization, observa-
tional, retrospective and multicentric study,
carried out in routine clinical practice. 419
patients with respiratory allergic disease,
who had received SLIT (an IR-standard-
ized extract) with a 4-day short conven-
tional titration scheme, were included.
Local and systemic reactions during the
build-up phase were extracted from exist-
ing patients records and classied accord-
ing the EAACI and MedDRA
classications.
Results: Two hundred and three patients
had received SLIT with pollen allergens
(48.4%), 205 with house dust mites
(HDM) allergens (48.9%) and four
patients with other allergens (0.9%). Sev-
enty one (16.9%) patients showed local
reactions (8.7% of doses) and nine (2.1%)
patients showed systemic reactions (0.7%
of doses) in the build-up phase. There were
no systemic reactions grade 2, 3 or 4 (EA-
ACI classication).
Conclusions: Due to its low incidence of
adverse reactions, sublingual immunother-
apy with IR-standardized extracts have an
excellent safety prole for the treatment of
allergic patients, even administered with a
rapid schedule for the build-up phase.
1660
A case of anaphylaxis after the first dose
of sublingual immunotherapy with house
dust mite
Park, H
Eulji University, School of Medicine, Department of
Pediatrics, Seoul, Republic of Korea
Background: The advantage of sublingual
immunotherapy (SLIT) is its friendly route
of administration and less frequent occur-
rence of severe side-effects. The most fre-
quently reported events were irritation of
the throat and oral itching. According to
the recent data, the number of side effects
seems to be dose-dependent. We report on
one cases of anaphylaxis after the rst dose
of SLIT with house dust mite drop.
Case: A 10-year-old girl, who suffered from
severe allergic rhinitis with perennial symp-
tom. She also suffered from asthma for
which a low dose inhaled corticosteroid was
used. Her allergy workup disclosed a posi-
tive skin prick test to D pteronyssinus, D
farina, birch, hazel tree, alder, dog, cat,
plantain, and acacia pollens. We started
with SLIT with Dermatophagoides (Pangr-
amin SLIT ALK-Abello, Madrid, Spain).
Fifteen minutes after the rst dose taken at
home, she experienced local irritation, lip
swelling, facial rash, rhinorrhea and cough.
She discontinued the treatment at home and
visited our clinic. Provocation test was per-
formed with the same drug(1.6 STU/ml,
1:625, 0.004 ug of Dp & Df). After 20 min,
the patient reported lip swelling, perioral
wheal, rash cough. The changes after provo-
cation test were described in table 1. She
recovered immediately after receiving a b2-
625

Table 1. For abstract 1660.
Pre-challenge
Clinical manifestation Mild wheezing
PEFR 310
BP(mmHg) 120/75
agonist inhalation and an oral antihista-
mine.
Results: The changes after provocation test
Conclusions: We would like to attract the
clinicians attention to potential systemic
side effects with SLIT despite the good
safety prole generally reported for this
procedure. We stress to have the rst dose
of SLIT taken in a doctors ofce with an
observation period of at least 30 min.
1661
Use of omalizumab prior to
immunotherapy in a patient with a
severe asthma allergic to mite
de Blay, F; Bloch, Y; Purohit, A; Darouaz, S
University Hospital of Strasbourg, Allergy Division,
Strasbourg, France
Omalizumab has been used in conjunction
with immunotherapy
We report the case of a 41-year-old male
patient with a severe allergic asthma to
mite. He had been suffering from rhinitis
and asthma since the childhood. The diag-
nosis of allergy to mite was based on a
positive prick test (Stallergenes) and spe-
cic IgE, (Phadia) to Dpt and Df 48.9 kU/
l and 44.3 kU/l respectively. The total IgE
levels was 1000 kU/l. In January 2005,
XOLAIR (225 mg every 2 weeks) was
introduced. Within 4 months, the patient
improved and on year after, he stopped his
oral corticosteroid treatment with no
asthma exacerbation. The inhaled corticos-
teroids treatment was decreased by 50%.
In 2008, on patients request, we began
sublingual immunotherapy to mite (Staller-
genes 10 drops a day, 300IR) with a good
tolerance. In February 2009, XOLAIR
was stopped and 2 years later, his asthma
is still controlled without increase of treat-
ment. In conclusions, this case underlined
the possibility of having a long term
improvement of severe allergic asthma to
mite by using XOLAIR in the beginning
and to continue having a prolonged ef-
cacy without XOLAIR under Immuno-
therapy to mite alone.
626
Poster Session 72 New data on efcacy, safety and mechanistic aspects of immunotherapy
Post -challenge
Lip swelling, perioral
wheal & rash, wheezing aggravation
270 (13%)
120/70
1662
Clinical experience in pretreatment of
allergen immunotherapy with oma-
lizumab in patients with severe uncon-
trolled asthma
Kuprys-Lipinska, I; Tworek, D; Korczynska, P; Antczak-
Marczak, M; Kuna, P
Medical University of Lodz, Internal Medicine, Asthma
and Allergy, Lodz, Poland
Omalizumab is the monoclonal anti-IgE
antibody used for treatment of severe aller-
gic asthma. Recent few reports have shown
the effectiveness of omalizumab in pretreat-
ment of allergen immunotherapy (IT) in
patients with venom or inhaled allergy. We
present three cases of patients with severe
allergic asthma pretreated with omalizumab
before induction of specic IT. First patient
was 33-year-old male with severe atopic
dermatitis and uncontrolled asthma, who
did not respond to standard treatment.
Clinical evaluation revealed serum IgE level
of 357 IU/ml and positive skin prick tests
and sIgE to house dust mites, cats and dogs
fur. All allergens were clinically signicant.
Omalizumab therapy was performed for
6 months (300 mg sc every 2 weeks) until
signicant improvement in asthma and ato-
pic dermatitis and stabilization of clinical
state. Then IT was started with Dermato-
phagoides and cat/dog (50%/50%) vac-
cines. No adverse events were observed due
to omalizumab or IT. After 5 years of IT
both asthma and atopic dermatitis are still
well controlled. The second patient, 65-
year-old female with severe asthma, resis-
tant to conventional treatment, with serum
IgE level 308 IU/ml and allergy to house
dust mites, tree pollens, dogs and cats fur.
After one year of treatment with oma-
lizumab (225 mg sc every 4 weeks) we
observed signicant improvement of
asthma and then decided to start allergen
immunotherapy with tree pollens and Der-
matophagoides mix, the omalizumab treat-
ment was continued. No adverse events
related to the treatment were occurred.
After 3 years of combined treatment
asthma is well controlled. The third patient
is 15-years-old girl suffering from asthma
with frequent severe exacerbations after
allergen exposition (house dust) and due to
the infections. Total IgE was 408 IU/ml,
and sIgE to Dermatophagoides in IV class.
The rst attempt of IT induction with Der-
Allergy 66, Suppl. 94 (2011): 482642  2011 John Wiley & Sons A/S
matophagoides mix vaccine nished unsuc-
cessfully due to the recurrent asthma
exacerbations. The second IT induction
was started after pretreatment with 8 weeks
therapy with omalizumab (450 mg sc every
4 weeks). The ongoing induction is per-
formed using cluster scheme and the oma-
lizumab treatment is continuing until the
end of induction phase.
1663
Omalizumab-related adverse events in
clinical trials of allergic rhinitis: a system-
atic review and meta-analysis
Tsabouri, S1; Tseretopoulou, X2; Priftis, K3; Ntzani, E2
1
Ioannina University Hospital, Department of
Paediatrics, Ioannina, Greece; 2Ioannina University,
Department of Hygiene and Epidemiology, Ioannina,
Greece; 3Athens University, Department of Paediatrics,
Athens, Greece
Background: Allergic rhinitis (AR) is a
common IgE-mediated inammatory disor-
der with limited mortality compared to
other allergic disorders, such as asthma,
but considerable impact on quality of life.
Treatment with subcutaneous omalizumab,
already proven effective in asthma, has
been also used in moderate to severe AR.
As omalizumab is a recombinant
humanized monoclonal anti-IgE antibody,
the omalizumb safety prole is of particu-
lar interest in the clinical setting of AR.
We performed a systematic review and
meta-analysis of published randomized
clinical trials (RCT) to assess the reported
safety of subcutaneous omalizumab in the
treatment of AR.
Methods: We searched for randomized tri-
als assessing the efcacy and safety of
omalizumab in PubMed and the Cochrane
Library. Two investigators independently
extracted the data and reached consensus
on all extraction items. For each report we
gathered information on study characteris-
tics and demographics. For each trial com-
paring omalizumab versus placebo, we
captured adverse events occurring in the
compared arms and denitions provided
thereof. We also captured information sep-
arately for any adverse events, specic
adverse events, serious adverse events and
drug-related adverse events. We synthe-
sized data across studies under a xed
effects or random effects model as appro-
priate. Heterogeneity was assessed using
the Q statistic and I2.
Results: Nine RCTs provided quantitative
evidence on adverse events. Adverse events
denitions varied across studies, while a
limited number studies did not report on
serious adverse events. Seven studies pro-
vided detailed data on the occurrence of
any adverse event on a cumulative sample
Poster Session 72 New data on efcacy, safety and mechanistic aspects of immunotherapy
size of 1,643 AR patients. The overall inci-
dence and pattern of any adverse event
were similar in the compared groups, with-
out reaching statistical signicance, but
with signicant heterogeneity present (ran-
dom effects RR 1.05; 95% CI 0.92-1.19;
P = 0.48; I2 57%). All reported adverse
events were of a mild degree and disap-
peared without any additional treatment.
No anaphylactic shock occurred, no anti-
omalizumab antibodies were detected in
any patient, and no signicant other
immune-complex mediated disorders were
observed.
Conclusions: The accumulated published
randomized evidence indicates that subcu-
taneous omalizumab was generally well tol-
erated in AR patients, although the
currently available evidence cannot exclude
less common, more serious adverse events.
1664
Italian survey on the safety of ultrashort-
term specific immunotherapy with pollen
allergoids adjuvanted by monophospho-
ryl lipid A (issue study)
Mariangiola, C1; Crivellaro, M1; Senna, G2
1
University of Padova, Allergology, Padova, Italy;
2 Gang, I; Jung, J; Kim, S
Azienda Ospedaliera Universitaria di Verona,
Allergology, Verona, Italy
Background: The clinical efcacy of aller-
gen specic immunotherapy in the treat-
ment of allergic respiratory diseases is well
documented. However its main drawbacks
are the risks of inducing systemic reactions.
A recently developed immunotherapy with
pollen allergoids adjuvanted by monophos-
phoryl lipid A requires a very ultrashort
pre-seasonal schedule of only four injec-
tions (uSCIT-MPL-4) and it has been
proved safe in controlled trials. However
its safety has not yet widely evaluated in
real life by post-marketing surveillances.
Objective: To prospectively assess the
safety of uSCIT-MPL-4 in a one-year,
multi-center, real-life study.
Methods: Twelve Italian allergology
departments were asked to record the clini-
cal features of systemic reactions due to
uSCIT-MPL-4. Vaccines were prescribed
according to the international guidelines.
Systemic reactions (SR) were graded
according to the EAACI recommendations
and were classied as immediate
(<30 min) or delayed (>30 min).
Results: A total of 788 patients (469 males
and 319 females; age range: 668 years)
Allergy 66, Suppl. 94 (2011): 482642  2011 John Wiley & Sons A/S
received uSCIT-MPL-4 (42.2% for grass,
28% for parietaria, 12.3% for ragweed and
17.4% for other pollens such as birch, arte-
misia and olea) and 5.8% of subjects
received more than one extract. A total of
3.902 injections were given. Fifty three
patients (6.73%) presented local reactions.
Overall, in the 4.7% of patients a total num-
ber of 58 SR (from grade zero to grade two)
were observed; reactions of grade more than
two were not observed. The 1.9% of
patients had SR of grade =>1 (CI: 0.01
0.03). No signicant differences in the recur-
rence of SR between different allergen
Immunotherapy were observed (P > 0.05).
Conclusionss: The rate of systemic reac-
tions in this prospective study was very
low demonstrating that uSCIT-MPL-4 is a
safe treatment with an acceptable risk/ben-
et ratio when prescribed and carried out
according to recommendations. In fact no
severe reactions were registered and epi-
nephrine was not used in any case.
1665
Clinical effect of sublingual immunother-
apy in allergic rhinitis patients
Gachon Medical School, Gil Medical Center,
Otolaryngology, Incheon, Republic of Korea
Background: Recently the efcacy of sub-
lingual immunotherapy (SLIT) in patients
with allergic rhinitis has been reported in
many studies. However, most of them has
used only one allergen, house dust mite or
grass pollen by using one kind of product.
In our study, we have used some kinds of
allergens from different products to evalu-
ate the early effect (6 month) of SLIT.
Methods: Ninety-one patients started the
SLIT from March 2008. The symptom
improvement, satisfaction rate, adverse
effects, combined diseases were investi-
gated. Types of SLIT products were Pangr-
amin for 24 patients, SLI-one for ve
patients, Stalora for 62 patients. Types of
allergens were Dermatophagoides pteronys-
sinus and Dermatophagoides farinae in 82
patients, others (Fungus, Tree, Grass) for
nine patients.
Results: The mean duration of SLIT was
7 months. Fifty four of the 91 patients
continued SLIT for more than 6 months
(59.3%). According to the ARIA classica-
tion, moderate to severe persistent patients
are 75%. Associated allergic diseases were
asthma in 14.7% and atopic dermatitis in
27.9%. The symptom score was improved
in 45.7% and the satisfaction rate was
73.5%. The decrease rate of medication
use was 51.4%. The incidence of adverse
effect was 25%. The most common cause
of discontinuing SLIT is side effect
(28.6%).
Conclusions: Sublingual immunotherapy
could be alternative treatment of allergic
rhinitis. However, further study about the
long term effects of SLIT should be done.
1666
Specific immunotherapy improves the
clinical response in patients with allergic
rhinoconjunctivitis
Arsovski, Z1; Gavrilovski, M1; Dokic, D2; Busljetic, K2;
Kaeva, B2
1
Clinic of Pulmonology and Allergy, Allergy
Department, Skopje, Macedonia, FYROM; 2Clinic of
Pulmonology and Allergy, Skopje, Macedonia, FYROM
Background: Specic immunotherapy (SIT)
has a signicant role in the treatment of
allergic rhinitis and allergic conjunctivitis.
The aim of this study was to evaluate the
effect of specic immunotherapy on grass-
pollen induced rhino-conjunctivitis.
Methods: We have evaluated the effect of
standard regiment of SIT on the pollen
allergic rhinoconjunctivitis. Twenty-six
patients (age range 1844 years) with posi-
tive history for more than 2 years and
skin-prick test 5 mm underwent conjunc-
tival provocation tests before and after
1 year of SIT. Clinical severity of nasal
and conjunctival symptoms during the sea-
son was assessed by 4-point arbitrary rat-
ing scale from 0 to 3. Conjunctival
provocation were done out of the season
until allergic symptoms occurred achieving
the allergen threshold dose (ATD).
Results: After 1 year of SIT we found
reduction in clinical symptoms of allergic
conjunctivitis (burning, itching, lacrimation
and hyperemia) P < 0.05 as well as reduc-
tion in clinical symptoms of allergic rhinitis
(secretion, irritation, itching and nasal
blockade) P < 0.01. Signicantly higher
allergen doses in provocation test (out of
season) were tolerated after 1 year of per-
formed SIT, reaching new ATD.
Conclusions: Grass pollen SIT reduces clin-
ical symptoms of allergic rhinoconjunctivi-
tis and modies the inammatory response
after specic allergen challenge.
627

Patient recorded outcomes and other paraclinical aspects in
1667
Adherence to IT: the patients point of
view
Bruno, M1; Modiano, A2; Cavazzuti, L2
1
Lofarma S.p.A., Medical Department, Milan, Italy;
2
Modena District, Specialized Assistance Department,
Modena, Italy
Background: Adherence is usually dened
as the extent to which patients use their
medication as prescribed. Adherence may
be considered a major concern in immuno-
therapy (IT), as in other long term thera-
pies. A relevant problem in IT is that it
requires substantial commitment by
patients, particularly in case of sublingual
IT (SLIT), since the treatment is self-
administered. Anyway, the main factors
inuencing adherence to IT regardless if
sublingual or injective (SCIT) are still in
discussion. Purpose of the study was to
evaluate the main factors inuencing
adherence to SLIT and SCIT according to
the patients point of view.
Methods: We handed out a questionnaire
to 38 patients undergoing IT, 8 SCIT and
30 SLIT, from 1 to 5 years. All patients
were informed about the duration, side
effects, delayed relief of allergy symptoms
and other features concerning IT. This
allowed them to perform a responsible
choice. Participants were 13 women and 25
men, average age 32 years (range 1460)
sensitized to grass or birch pollen or mites,
with moderate persistent rhinoconjuntivitis
and mild intermittent asthma. We evalu-
ated their replies to a 13 items multiple
choice questionnaire, each question having
a choice among 6 scores (from 1-make
worse to 6-improves very much), evaluat-
ing the impact on adherence to IT. The
questions concerned cost, reimbursability,
long lasting therapy standing, latency of
symptoms relief, condence in prescribing
doctor, other doctors opinions, IT side
effects, control visits, SLIT tablets, SLIT
drops, SCIT, therapy efcacy perception,
daily therapy intake. The last question was
a one choice question, concerning which
item was the most important for the
patient adherence to IT.
Results: Patient perception of clinical ef-
cacy clearly emerged as the most important
628
Poster Session 73
immunology
factor in improving adherence to IT, fol-
lowed by the condence in the prescribing
doctor. Tablet SLIT was the most appreci-
ated form of IT. Cost and reimbursement,
on the contrary, did not affect signicantly
adherence, as well as side effects.
Conclusions: Perceived efcacy is the major
issue inuencing adherence. There is not a
signicant difference in adherence between
SLIT and SCIT, although SLIT is consid-
ered more acceptable.
1668
Noticeable placebo response in clinical
studies investigating efficacy of different
preparations for subcutaneous specific
immunotherapy
Lehnigk, U; Kettner, J; Haefner, D; Cromwell, O;
Narkus, A
Allergopharma Joachim Ganzer KG, Reinbek, Germany
Background: Randomised, double-blind,
placebo controlled (RDBPC) studies are
currently the recommended standard of
clinical trials. The outcome of these trials
is affected by the placebo response which
is driven by different mechanisms and
inuenced by a variety of factors e.g. inva-
siveness and duration of therapy or
patients attitude of expectation. Only clini-
cal trials including a baseline exposure per-
iod provide the possibility to precisely
investigate this effect in SIT. In past stud-
ies, only to a lesser extent baseline assess-
ments were included.
Methods: We analysed ve randomised,
double-blind, placebo controlled clinical
trials including a baseline period, designed
to assess efcacy and safety of SCIT in
patients with allergic rhinitis (AR), allergic
to grass pollen, birch pollen or mites, using
the full analysis sets (FAS). In a total of
415 placebo treated patients the median
change relative to baseline of the area
under the curve (AUC) of the symptom
medication score (SMS) after 2 years of
placebo therapy was calculated. Evaluation
periods covered 21 days for birch pollen,
42 days for grass pollen as well as 28 days
for house dust mite (HDM) allergy. To
obviate the possibility that different pollen
exposure during the seasons inuenced the
Allergy 66, Suppl. 94 (2011): 482642  2011 John Wiley & Sons A/S
results, the AUCs of birch and grass pollen
grains/m3 during the evaluation periods
were checked.
Results: Birch pollen allergic placebo trea-
ted patients with AR (n = 216) demon-
strated an improvement of the AUC of
median SMS of 21.5%, grass pollen aller-
gic placebo treated patients (n = 87) an
improvement of 22.5%. Patients belonging
to the placebo group with HDM allergy
(n = 112) displayed an even exceeded
improvement by 35.5% of their SMS com-
pared to baseline. Pollen counts of the sea-
sonal allergens were comparable for the
analysed periods in 2 years.
Conclusion: The results of different
RDBPC clinical trials with a baseline per-
iod showed that the inuence of subcutane-
ous placebo therapy is in the range of 22
36% in SIT studies. Further investigations
have to show if diverse application routes
e.g. the sublingual route the placebo
response to the same extent.
1669
Patients compliance towards high-dose
hypoallergenic and unmodified SCIT
treatment schedules is superior to
sublingual immunotherapy
Egert-Schmidt, A; Martin, E; Mueller, J; Schulte, M;
Thum-Oltmer, S
Allergopharma Joachim Ganzer KG, Reinbek, Germany
Background: To benet from long-term
efcacy and disease-modifying effect of
SIT, it is crucial to achieve high compli-
ance rates and treatment adherence over a
time period of 35 years. According to
guidelines, expected non-compliance is
even a contraindication for SIT.
Methods: Aim of this study was to com-
pare patients compliance when receiving
SIT with different Allergopharma prepara-
tions for SCIT ((high-dose hypoallergenic
or unmodied), (perennial or preseasonal))
and SLIT, based on patients data from
the German market. Patients who received
rst prescription for SCIT in 2005 and
SLIT in 2007 were included, depending on
the launch date of the respective pre-
parations. To identify patients compli-
ance, completed treatment years were
Poster Session 73 Patient recorded outcomes and other paraclinical aspects in immunology
Table 1. For abstract 1669.
High-dose hypoallergenic pollen SCIT perennial
High-dose hypoallergenic pollen SCIT preseasonal
High-dose hypoallergenic mite SCIT
Unmodied pollen SCIT perennial
Unmodied mite SCIT
Pollen SLIT
determined. A completed treatment year
was dened by the required number of pre-
scribed allergen preparations according to
the dosage scheme of each preparation, as
described in the respective SmPC.
Results: The table shows the compliance
rates of different SIT preparations. 58% of
patients treated with high-dose hypoaller-
genic pollen SCIT have completed 3 treat-
ment years when administered perennially.
Patients under SCIT with a high-dose
hypoallergenic mite preparation showed
slightly lower compliance rates. For
unmodied SCIT preparations the compli-
ance rates for 3 treatment years were
41% for pollen and 49% for mites. The
compliance rate for pollen SLIT was
shown to be lowest: only 16% of patients
took the medication for at least 3 years, as
recommended.
Conclusion: Based on patients data from
the German market, the perennial treat-
ment with high-dose hypoallergenic pollen
SCIT seems to be favourable to achieve
high compliance rates. In general, SCIT
preparations (hypoallergenic or unmodi-
ed) are superior to SLIT preparations
regarding compliance issues and treatment
adherence.
1670
Patients compliance and adherence with
solid versus liquid forms of sublingual
immunotherapy in the management of
allergic rhinitis
Komarla, N1; Kanni, P1; Bhathe, S2
1
BAIRI, Allergy, Bangalore, India; 2BAIRI, Allergy Dept,
Bangalore, India
Background: Slit-drops is known for its
efcacy in the management of Allergic
Rhinitis. Drops has limitations which were
expressed by storage, transportation, carry-
ing while distance traveling, storing in vil-
lage environment and for those who stayed
at hostels. It is not very friendly among
children, where they cannot retain more
than stipulated time and majority will spit
since it is not nature friendly.
Methods: BAIRI research team initiated to
develop tailor made allergen extract of
House Dust mite (Dermatophagoides fari-
nae and Dermatophagoides pteronysinus) in
solid physical form of thin lm known as
STRIPE. The Stripe were dispensed to
Allergy 66, Suppl. 94 (2011): 482642  2011 John Wiley & Sons A/S
3 years 2 years 1 year
58% 85% 100%
28% 49% 100%
49% 63% 74%
41% 58% 65%
48% 63% 66%
16% 26% 54%
Allergic Rhinitis subjects and compared to
drops to generate comparative compliance
among two groups in terms of friendly use,
dened dosage delivery, storage and com-
pliance with efcacy and adherence. Sixty
seven patients were on Slit-stripe and 48
patients were on Slit-drops, having symp-
toms from 1 year to 10 years.
Results: The delivery of the dosage form in
Stripe were pre xed by unit. The drops
dosage varies with volume and count since
the container and the delivery unit is under
the inuence of pressure and the person
who operates, which was not uniform from
dose to dose. Allergic rhinitis symptoms
were well controlled over stripe compara-
ble to drops and its efcacy was well
recorded and comparable to drops. Moth-
ers of children were happy and friendly in
use since the stripe is easily stick to base of
the tongue and retains for few minutes till
it dissolves. The adherence and follow up
was found more with Stripe than drops.
The storage and carrying while traveling
need no special carrier like ice pack or the
thermos ask. The shelf life is more with
stripe than with drops.
Conclusion: SLIT in the form of STRIPE
were well documented and found more
benecial and with meritorious compliance.
It was well comparable to drops in long
term management of Allergic Rhinitis by
SLIT STRIPE.
1671
A comparative health economic evalua-
tion of the SQ-standardised grass allergy
immunotherapy tablet and subcutaneous
immunotherapy in the treatment
of grass pollen induced allergic
rhinoconjunctivitis
Ronborg, S1; Svendsen, U2; Micheelsen, J3; Ytte, L4;
Riss Treloggen, J5; Andreasen, J6; Ehlers, L7
1
The Pulmonology and Allergy Clinic of Copenhagen,
Copenhagen, Denmark; 2Bispebjerg Hospital,
Copenhagen, Denmark; 3Private ENT practice, Denmark;
4
General practitioner, Humlebaek, Denmark; 5ALK-
Abello, Denmark; 6ALK-Abello, Copenhagen, Denmark;
7
Aalborg University, Denmark
Background: Allergic rhinoconjunctivitis
(ARC) constitutes a large burden for soci-
ety. The disease prevalence is increasing
and app. 2025% of the European popula-
tion suffers from respiratory allergies
including ARC. The majority of patients
are treated with symptomatic pharmaco-
therapy; however a large proportion
remain uncontrolled despite the use of such
treatments. Specic immunotherapy (SIT)
is the only treatment documented to target
the underlying allergic disease and activate
immunomodulatory mechanisms leading to
a sustained effect after completion of treat-
ment. Based on Danish societal and health
care perspective, this analysis shows the
economic consequences if patients with
grass pollen induced ARC uncontrolled on
symptomatic medications are treated with
a grass allergy immunotherapy tablet
(AIT) instead of the currently established
clinical practice of subcutaneous immuno-
therapy (SCIT).
Methods: A cost-minimisation analysis
(CMA) was applied comparing the SQ-
standardised grass AIT (Grazax, Phleum
pratense, 75 000 SQ-T/2800 BAU, ALK,
Denmark) with SCIT (Phleum pratense,
alum-absorbed depot extract, maintenance
dose of 100 000 SQ-U, ALK, Denmark).
The choice of CMA was based on a review
publication and a meta-analysis, which
concluded that the clinical effect of AIT is
similar in magnitude to that observed for
SCIT in grass pollen induced ARC patients
with similar disease severity. Health care
utilisation was measured in physical units
based on national guidelines, literature
reviews and expert opinion. The valuation
in unit costs was based on drug tariffs,
physician fee structures and wage statistics.
Results: Treating ARC patients with the
grass AIT instead of SCIT results in a sig-
nicantly reduced number of physician vis-
its leading to a reduction in direct
treatment costs, patient-borne costs and
indirect costs. The expected reduction in
direct treatment costs is 1289 per patient
during a treatment course. If the societal
perspective is included, the reduction in
cost is 3723 per patient. The sensitivity
analysis conrms the robustness of the
results.
Conclusion: In patients with grass pollen
induced ARC, this cost-minimisation anal-
ysis shows that allergy treatment with the
SQ-standardised grass AIT reduces both
the direct and indirect treatment costs
compared with SCIT. Thus, the SQ-stan-
dardised grass AIT is a cost-saving alterna-
tive to SCIT both from a societal and
health care perspective.
629

1672
Quality of life improvement in patients
suffering from grass pollen-related rhino-
conjunctivitis and asthma: 1 year follow-
up of a 3 year ultra-short course subcuta-
neous immunotherapy containing mono-
phosphoryl-lipid A
Rabe, U1; Fiedler, G1; Piller, M2
1
Germany; 2Bencard Allergie, Munich, Germany
Background: Allergic diseases interfere sig-
nicantly with many aspects of daily life
and have a great impact on the quality of
life of the affected patients. After three
courses of SCIT with Monophosphoryl-
Lipid A (MPL) patients show signicant
improvements in quality of life parameters
(presented previously). In this extension
study the further improvement on the qual-
ity of life of patients with allergic rhinitis
and/or asthma was investigated.
Methods: A total of 56 out of 73 patients
with grass pollen-related rhinoconjunctivi-
tis and/or asthma who had completed the
third course of the previous multicentre,
non-interventional trial were included in a
long-term evaluation of their disease
related quality of life. Yearly Rhinitis and
Asthma Quality of Life (RQLQ and
AQLQ) questionnaires were lled in by the
patients.The score for both questionnaires
ranged from 0 (not affected at all) to 6
(extremely affected). All patients had com-
pleted the same SCIT (4 injections) con-
taining a mixture of grass pollen allergoid
and MPL for three consecutive years n-
ished one year before this evaluation.
Results: The total mean score (longitudinal
model) for the patient-documented RQLQ,
which had shown a statistically signicant
improvement following the three treatment
courses from 2.6 score points to 1.6
(P < 0.0001), shows sustained efcacy
with only minor changes from 1.6 score
points to 1.7 1 year after the third treat-
ment course. The AQLQ total mean score
(longitudinal model) shows similar changes
from 2.1 score points to 1.1 (P < 0.0001)
during the three year treatment and
increases to 1.2 score points one year after
the treatment and therefore reects sus-
tained efcacy for asthma symptoms as
well. Comparing to the last pollen season,
the doctors assessment of the therapy suc-
cess showed positive effects for the rhino-
conjunctivitis-symptoms in 81% (2007),
81% (2008) and 97% (2009) of patients. In
the follow-up year (2010) rhinoconjunctivi-
tis-symptoms improved further in 55%,
remained unchanged in 41% and worsened
in 2% of patients.
Conclusion: Quality of life questionnaires
for rhinoconjunctivitis and for asthma
show sustained efcacy in the rst year
without immunotherapy after three courses
630
Poster Session 73 Patient recorded outcomes and other paraclinical aspects in immunology
of treatment with MPL-adjuvanted grass
pollen SCIT.
1673
Global Initiative for Asthma classification
of asthma control derived from the
Asthma Control Questionnaire data
Virchow, J1; Bufe, A2; Seitzberg, D3; Bateman, E4
1 of FEV1 contributes little to the asthma
University of Rostock, Department of Pneumology,
Rostock, Germany; 2Ruhr-Universitat Bochum,
Experimentelle Pneumologie, Bochum, Germany;
3
Research & Development, ALK, Hrsholm, Denmark;
4
University of Cape Town, Department of Medicine,
Division of Pulmonology, Cape Town, South Africa
Background: The Asthma Control Ques-
tionnaire (ACQ) is a useful scientic tool
for assessing levels of asthma control in
clinical trial settings. However, clinicians
may be unfamiliar with the ACQ scores.
Therefore we aimed to translate ACQ
scores into the Global Initiative for
Asthma (GINA) treatment guideline termi-
nology for asthma control, i.e. con-
trolled, partly controlled or Rabe, U1; Fiedler, G1; Piller, M2
uncontrolled to make them more easily
interpretable. This may be useful both
when understanding clinical trial data, and
interpreting it for clinical use. Data from a
large, randomised, double-blind, placebo-
controlled, trial with three doses of the
house dust mite (HDM) allergy immuno-
therapy tablet (AIT) (ALK, Denmark) was
used.
Methods: The trial population comprised
604 subjects, aged 14 years or older with
evidence of HDM-induced mild to moder-
ate persistent asthma according to the
GINA 2006 guideline and with daily use of
low-medium dose inhaled corticosteroid
(ICS). Prior to randomisation, the use of
ICS was standardised and tapered to the
lowest dose providing adequate asthma
control. ACQ scores were obtained at ran-
domisation. Post-hoc an algorithm was
constructed to translate the seven ACQ
questions into GINA terminology.
Results: At randomisation, 21%
(N = 127) of the trial population had con-
trolled asthma according to GINA termi-
nology, with a mean ACQ score of 0.24
[CI: 0.200.28] and mean FEV1 of 96% of
predicted value. Another 69% (N = 412)
had partly controlled asthma, with a mean
ACQ score of 0.86 [CI: 0.820.90] and a
mean FEV1 of 94%. Finally, 10%
(N = 60) had uncontrolled asthma, with a
mean ACQ score of 1.54 [CI: 1.411.66]
and a mean FEV1 of 90%. In the trial
population overall, the HDM AIT was
well-tolerated and efcacious in terms of
ICS reduction.
Conclusion: From the ACQ it is possible
to classify the patients levels of asthma
control into the GINA 2009 denitions of
Allergy 66, Suppl. 94 (2011): 482642  2011 John Wiley & Sons A/S
controlled, partly controlled or
uncontrolled. The selection of subjects
with HDM-induced mild to moderate per-
sistent asthma and low-medium ICS use
resulted in a trial population comprising
69% of subjects with partly controlled
asthma according to GINA terminology
and 10% with uncontrolled asthma. The
data conrmed that lung function in terms
control classication. Our data suggest that
in patients with partly or even uncontrolled
asthma, whose lung function is not severely
impaired, treatment with HDM AIT may
be well tolerated.
1674
Quality of life improvement in patients
suffering from tree pollen-related rhino-
conjunctivitis and asthma: 1 year follow-
up of a 3 year ultra-short course subcuta-
neous immunotherapy containing mono-
phosphoryl-lipid A
1
Germany; 2Bencard Allergie, Munich, Germany
Background: Allergic diseases interfere sig-
nicantly with many aspects of daily life
and have a great impact on the quality of
life of the affected patients. After three
courses of SCIT with Monophosphoryl-
Lipid A (MPL) patients show signicant
improvements in quality of life parameters
(presented previously). In this extension
study the further improvement on the qual-
ity of life of patients with allergic rhinitis
and/or asthma was investigated.
Methods: A total of 76 out of 112 patients
with tree pollen-related rhinoconjunctivitis
and/or asthma who had completed the
third course of the previous multicentre,
non-interventional trial were included in a
long-term evaluation of their disease
related quality of life. Rhinitis and Asthma
Quality of Life (RQLQ and AQLQ) ques-
tionnaires were lled in by the patients at
three time points: retrospectively with
regard to the last pollen season; during,
and at the end of the pollen season one
year after the third treatment course. The
score for both questionnaires ranged from
0 (not affected at all) to 6 (extremely
affected). All patients had completed the
same SCIT (four injections) containing a
mixture of tree pollen allergoid and MPL
for three consecutive years nished one
year before this evaluation.
Results: The total mean score (longitudinal
model) for the RQLQ, which had shown a
statistically signicant improvement follow-
ing the three treatment courses from 3.1 to
1.9 (P < 0.0001), shows sustained efcacy
from the patients point of view with only
minor changes from 1.9 to 2.0. The AQLQ
Poster Session 73 Patient recorded outcomes and other paraclinical aspects in immunology
total mean score (longitudinal model)
shows similar changes from 2.2 to 1.2
(P < 0.0001) during the three year treat-
ment and increased to 1.3 one year after
the treatment and therefore reects sus-
tained efcacy in the patients view of
asthma symptoms as well. Comparing to
the last pollen season, the doctors assess-
ment of the therapy success showed posi-
tive effects for the rhinoconjunctivitis-
symptoms in 81% (2007), 80% (2008) and
91% (2009) of patients. In the follow-up
year (2010) rhinoconjunctivits-symptoms
improved further in 51%, remained
unchanged in 40% and worsened in 9% of
patients.
Conclusion: Quality of life shows sustained
efcacy in the rst year without preseason-
al immuno therapy after three courses of
SCIT treatment This was shown for the
RQLQ and AQLQ questionnaires.
1675
The relationship between allergic disease
severity and quality of life in patients
treated with sublingual immunotherapy
tablets for house dust mite-induced rhini-
tis
Demoly, P1; Worm, M2; Fokkens, W3; Tabar, A4;
Nguyen, H5; de Beaumont, O5; Melac, M5; Montagut,
A5; Bergmann, K6
1 1676
University Hospital of Montpellier, Montpellier, France;
2
Allergy-Centre-Charite, Charite - Universitatsmedizin
Berlin, Germany; 3Department of Otorhinolaryngology,
AMC, Amsterdam, The Netherlands; 4Department of
Allergy, Complejo Hospitalario de Navarra, Pamplona,
Spain; 5Stallergenes SA, Antony, France; 6Allergy-
Centre-Charite, Charite Universitatsmedizin Berlin,
Berlin, Germany
Background: The aim of this study was to
determine the relationship between disease
severity and quality of life of 509 patients
included in a 2 year multinational, ran-
domized, double-blind, placebo-controlled
phase II/III study.
Methods: Patients were randomized to a
placebo and two active groups, receiving
either 300 or 500IR sublingual tablets dur-
ing the treatment period. The rhinocon-
junctivitis quality of life questionnaire
(RQLQ) developed to measure the effects
of patients nasal and ocular rhinoconjunc-
tivitis symptoms on the quality of life was
one the trials secondary endpoints. Post-
hoc analyses on patients without con-
founding sensitizations (N = 128) were
performed to evaluate the potential rela-
tionship between disease severity and treat-
ment effect in the 300IR group; centres
were divided into three subgroups of equal
size (tertiles) according to their mean aver-
age adjusted symptom score in placebo
group (AAdSS, a score which adjusts
symptom severity for rescue medication
use).
Allergy 66, Suppl. 94 (2011): 482642  2011 John Wiley & Sons A/S
Results: At the end of the treatment year,
the low, medium and high level of symp-
toms tertiles of the 300IR group showed
relative LS Means differences in the overall
RQLQ score (versus. placebo) of 0.3%,
)35.9% and 36.8%, respectively. In those
tertiles groups, the scores of sleep domain
were respectively 1.7%, 30.3% and
48.0%. In high level of symptoms patients,
the eye symptoms and sleep domains gen-
erated the highest relative difference in LS
Means during the treatment period (
53.1% and 48.0%, respectively). The
results showed that the more severe was
the disease, the greater was the effect on
the improvement of sleep. The benecial
effect of HDM-SLIT on the quality of life
increased with the severity of the disease.
Conclusion: Quality of life is improved
upon HDM SLIT and interestingly most
pronounced improvement of QoL was
observed on the sleep domain with an
increasing benecial effect related the
severity of the disease. As the deterioration
of sleep is one hallmark on quality of life
being most troublesome for patients with
rhinitis these results suggest an additional
benet for HDM patients, besides clinical
efcacy.
Compliance and treatment discontinua-
tion rate of patients with house dust
mites-induced allergic rhinitis receiving
sublingual allergen immunotherapy
Malet, A1; Gutierrez, D2; Moreno, F3; Cumplido, J4;
Justicia, J5; Garcia, M5
1
Al.legro Centre, Barcelona, Spain; 2Hospital Puerta del
Mar, Cadiz, Spain; 3Clinica Dr. Lobaton, Cadiz, Spain;
4
Hospital Dr. Negrin, Las Palmas de Gran Canaria,
Spain; 5Stallergenes Iberica, Barcelona, Spain
Background: Allergen immunotherapy
(AIT) is a long term treatment able to
change the allergy course. AIT has adher-
ence problems of different magnitude
depending on the characteristics of studies.
The aim of the present study was to
increase knowledge about adherence to
treatment in patients allergic to house dust
mites (HDM) receiving sublingual allergen
immunotherapy (SLIT).
Methods: A retrospective observational
study was done in 53 Spanish allergy units.
Specialists pre-selected clinical history cases
of patients that had initiated SLIT for
HDM and who had a scheduled control
visit 12 months after initiating SLIT. When
the planned medical control visits were
done, study data was collected. Patients
were asked to answer a specic question-
naire about compliance and treatment dis-
continuation.
Results: A total of 380 (91.1%) of the 417
pre-selected cases participated in the study.
Compliance rate was 79.5% and treatment
discontinuation rate was 20.8%. 66.8% of
patients were compliant and continued
with treatment at the time of study. These
results varied in a statistically signicant
way according to age groups, children
being most adherent and adolescents the
least. Factors explaining the lack of full-
ment with SLIT also varied across ages.
Adherent and non-adherent patients dif-
fered in age, number of annual control vis-
its with an allergist and reduction in
symptomatic medication.
Conclusion: The problem of adherence to
SLIT may not be as severe, and may not
have the same reasons across different age
groups of allergic patients. This should be
taken into account when dening strategies
to improve adherence to AIT.
1677
Safety profile and compliance of co-sea-
sonal initiation of specific sublingual
immunotherapy with a perlingual spray
application
Dichmann, R
Medical Practice, Pulmonology, Allergology, Witten,
Germany
Background: Specic sublingual immuno-
therapy (SLIT) is an option for causal
treatment of IgE-mediated allergic diseases
and of increasing interest for many
patients. A suitable way of application and
a convenient treatment start even when the
pollen season is already ongoing should
enhance the acceptance and compliance by
the patients. Therefore the safety prole of
co-seasonal initiation of SLIT with a per-
lingual spray application was evaluated in
patients with allergic rhinitis due to tree
pollen.
Methods: Patients suffering from clinically
relevant allergy to tree pollen (28 patients,
17 female, 11 male, age range 1372 years)
started with SLIT during the appropriate
pollen season. An aqueous allergen extract
(SULGEN Spray) of birch pollen (10
patients) or a mixture of birch, elder and
hazel pollen (18 patients) was applied using
two puffs daily as the maintenance dose.
The safety prole during the rst weeks of
treatment as well as the patients compli-
ance has been assessed retrospectively.
Results: The treatment has been well toler-
ated, only three patients reported mild
itching in the mouth on the rst and sec-
ond treatment day, but later on no further
reactions occurred. All patients appreciated
the spray application and conrmed their
good compliance. All patients continued
treatment.
Conclusion: Co-seasonal initiation of SLIT
with a perlingual spray application has
631

been well tolerated and is a safe option for
specic immunotherapy resulting in good
compliance of the patients.
1678
Modern variation in specific immunother-
apy (SIT) as a concept for treating sol-
diers with allergies with regard to
foreign mission or changes in residency
Nebel, J; Jacob, R
Bundeswehrzentralkrankenhaus, ENT, Koblenz,
Germany
Introduction: The German Bundeswehr has
been involved in the 90s worldwide opera-
tions abroad. Currently, more than 5000
soldiers in Europe, Africa and Asia are sta-
tioned for several months. Added by fre-
quent changes of residence at home,
soldiers with allergies of the upper respira-
tory tract treated with immunotherapy
have difculties in preserving the continu-
ity of care maintained.
Procedure: Since 2006 at the Bundeswehr
Central Hospital Koblenz immunotherapy
is started by high speed saturation in one
day. With experience in many hundreds of
patients the current treatment regimens the
initial phase of SIT was reduced to 12 h.
In cases with prolonged interval between
injections, high speed re-saturation with
the same scheme as for the initial phase
was used.
Results: In comparison to the period
before 2006 this Koblenz Ultra-rush
scheme has resulted in many more soldiers
receiving and completing immunotherapy.
The great mobility of the soldier is no
longer an obstacle for the start of SIT. In
foreign missions, the treatment is contin-
ued by specialist army doctors. Although
treatment interruption in long missions or
difculties in supply, SIT was continued by
re-saturation scheme.
Conclusion: With modern immunotherapy
regimens using high-speed saturation ini-
tially and at intervals exceeded, most of
the highly mobile patients with allergies,
were successfully treated and the therapy
course completed. It needs to be examined
Table 1. For abstract 1680.
Weekly severe days: 0 1
n = 10335
Predicted RQLQ 0.79
score, mean
[95% CI] [0.77;0.81]
Difference in 0.00
RQLQ score at 0
weekly severe days
Odds-ratios Grass AIT 1.71
versus Placebo, mean
[95% CI] [1.56;1.89]
632
Poster Session 73 Patient recorded outcomes and other paraclinical aspects in immunology
whether the frequent exceed of predeter-
mined intervals of 4 weeks between injec-
tions, puts treatment outcomes at risk.
1679
Olfactory function in seasonal allergic rhi-
nitis can be improved by allergen-specific
immunotherapy
Klimek, L1; Glowania, A2; Stuck, B3; Hormann, K3; Pfaar,
O2
1
Center for Rhinology & Allergology, Wiesbaden,
Germany; 2University Hospital Mannheim, Center for
Rhinology & Allergology/ENT-Department, Wiesbaden,
Germany; 3University Hospital Mannheim, ENT-
Department, Mannheim, Germany
Backround: Olfactory dysfunction is a
common nding in patients suffering from
allergic rhinitis. However, little is known
about the effects of common therapeutic
methods such as allergen-specic subcuta-
neous immunotherapy.
Objective: To evaluate the inuence of a
pollen season on olfactory function and
the efcacy of a pre-seasonal immunother-
apy with subcutaneous injections of molec-
ular standardized allergens from grass and
rye pollen with drug treatment on demand
in comparison to a symptomatic drug
treatment alone on olfactory threshold,
identication, and discrimination.
Methods: A prospective controlled study
has been performed on 64 patients with
allergic rhinitis to grass and rye pollen that
had been randomized 1:1 to either SCIT
with symptomatic treatment on demand
(SCIT) or symptomatic medication, alone
(SM). A baseline season was compared to
a season after treatment. Olfactory func-
tion was evaluated at baseline, preseasonal-
ly, on day 3, 7, 14, 21 and 28 of the season
and after season using the SnifnSticks-
testing procedure for threshold, identica-
tion and discrimination. 12 non-allergic
volunteers served as controls.
Results: Out of season, patients and con-
trols performed equally in discrimination
and identication testing, but not in thresh-
old testing. No changes were found in the
controls, but a signicant decrease in thresh-
old and identication from the 7th day of
2 3
n = 1074 n = 422 n = 202
1.70 2.29 2.66
[1.66;1.74] [2.242.34] [2.59;2.73]
0.91 1.50 1.87
0.81 0.58 0.5
[0.72;0.92] [0.48;0.71] [0.38;0.67]
Allergy 66, Suppl. 94 (2011): 482642  2011 John Wiley & Sons A/S
the season in the allergics was noted in the
baseline season (P < 0.01). This decrease
was prevented in the SCIT group for the
entire period for discrimination and up to
day 21 for identication. In the SM group,
it was prevented until day 14 for identica-
tion and discrimination (P < 0.01).
Conclusions: Grass pollen allergics develop
olfactory dysfunction during natural aller-
gen exposure that might be better pre-
vented by SCIT with symptomatic
treatment then by symptomatic treatment
alone.
1680
Treatment with SQ-standardised grass
allergy immunotherapy tablet impacted
the number of days with low quality of
life
Andersen, J1; Bch, S1; Durham, S2
1
ALK, Hrsholm, Denmark; 2London, United Kingdom
Background: The quality of life of patients
with allergic rhinoconjunctivitis is often
impaired due to symptoms such as nasal
obstruction and rhinorrhea, frequently
resulting in sleep disruption. Purpose: to
investigate how the number of days with
severe symptoms (severe days) impacts the
rhinoconjunctivitis quality of life and how
the treatment with a grass allergy immuno-
therapy tablet (AIT) impacts the frequency
of severe days.
Methods: Data from a multicentre, double-
blind, randomised, placebo-controlled trial
of 5 years duration (3 years daily treatment
and 2 years follow-up) with the grass AIT
(Grazax, phleum pratense 75 000 SQ-T/
2800 BAU, ALK, Denmark), were used.
Six rhinoconjunctivitis symptoms (four
nose and two eye) rated on a 4-point scale
from 0 (no symptoms) to 3 (severe symp-
toms) were recorded on a daily basis.
Quality of life was assessed with the Juni-
pers rhinoconjunctivitis quality of life ques-
tionnaire (RQLQ) on a weekly basis in
terms of seven domains each rated on a 7-
point scale from 0 (not troubled/none of
the time) to 6 (extremely troubled/all of
the time). Post-hoc, the numbers of weekly
4 5 6 7
n = 121 n = 84 n = 56 n=30
2.93 3.14 3.33 3.49
[2.84;3.01] [3.04;3.25] [3.20;3.46] [3.28;3.71]
2.14 2.35 2.54 2.70
0.33 0.23 0.37 0.17
[0.22;0.49] [0.14;0.38] [0.21;0.65] [0.06;0.44]
Poster Session 73 Patient recorded outcomes and other paraclinical aspects in immunology
severe days in at least one of the six symp-
tom categories were merged with the
RQLQ replies the same week (12324 data
points) and analysed, using cubic B-splines
and logistic regression.
Results: The RQLQ score increased
(=lower quality of life) with the number
of weekly severe days. For example, the
RQLQ score was 3.7 times higher for sub-
jects with four or more weekly severe days
(mean score: 2.93) than for subjects with
no weekly severe days (mean score: 0. 79),
Table 1. Treatment with grass AIT signi-
cantly reduced the odds of having severe
days relative to placebo, P < 0.001, Table
1. The odds-ratio for grass AIT versus pla-
cebo for having severe days decreased with
the number of weekly severe days. For
example, the odds of having four or more
weekly severe days were three times lower
(odds-ratio = 0.3) in the group treated
with grass AIT.
Conclusion: Treatment with grass AIT sig-
nicantly reduced the odds of having days
with severe symptoms. Days with severe
symptoms had a negative impact on the
quality of life.
1681
Tolerability of the SQ-standardised grass
allergy immunotherapy tablet and
patient satisfaction with treatment in
children, adolescents and adults during
routine application
Gronke, C1; Wolf, H2; Schnitker, J3; Wustenberg, E4
1
Pediatrician, Ludwigsfelde, Germany; 2ALK-
Arzneimittel GmbH, Clinical Development, Wedel,
Germany; 3Institut fur angewandte Statistik GmbH,
Bielefeld, Germany; 4ALK-Arzneimittel GmbH, Medical
& Regulatory Affairs, Wedel, Germany
Background: The fast-dissolving SQ-stan-
dardised grass allergy immunotherapy tab-
let (AIT) became available for children
from 5 years of age in 2008 in Germany
and Austria. Objective of our study was to
record data about tolerability and patient
satisfaction in children, adolescents and
adults during routine use of grass AIT.
Methods: In a non-interventional, observa-
tional study 1761 patients (797 <18 years;
964 18 years) were treated with grass
AIT (GRAZAX; 75 000 SQ-T/2800
BAU, Phleum pratense, ALK, Denmark)
by 373 allergists in Germany and Austria
between November 2008 and January
2010. Tolerability of grass AIT was docu-
mented for the rst intake and the follow-
ing 69 months of treatment until the end
of the grass pollen season. At visits every
3 months compliance, convenience and tol-
erability were assessed. Symptoms and
symptomatic medication requirements for
the grass pollen seasons before and during
therapy were also assessed.
Allergy 66, Suppl. 94 (2011): 482642  2011 John Wiley & Sons A/S
Results: Adverse drug reactions (ADR)
were reported in 27.3% patients 18 years
and in 37.7% of patients <18 years. The
higher rate in the children and adolescents
group was due to a numerical higher fre-
quency of oral paraesthesia, throat irrita-
tion and oral pruritus as most common
ADR to grass AIT. Treatment was discon-
tinued due to non-adherence by 121
patients <18 years (15.2%) and
171 18 years (26.7%). Compliance rates
of >75% were observed in 89.9% of
patients, 96.1% of patients assessed the
application of grass AIT to be easy. Com-
pared with the previous grass pollen season
prior to grass AIT treatment, nasal symp-
toms were reported to be improved in
84.0% of patients <18 years and in 81.5%
of patients 18 years, and less symptomatic
medication was used by 92.6% of patients
<18 years and 86.9% of patients
18 years. Well-being was assessed to be
improved compared with previous years by
90.4% of patients. Proportion of patients
and physicians satised with grass AIT
treatment were both >95%.
Conclusion: The results of our study
regarding tolerability conrm the safety
prole observed in controlled clinical trials
with SQ-standardized grass AIT in chil-
dren, adolescents and adults. Treatment
satisfaction of the grass allergen tablet dur-
ing routine application was rated high in
combination with a good compliance.
1682
Pre-seasonal immunotherapy: is there
any change in the clinical efficacy and
impact on the quality of life between
3 years
Goksel, O1; Aydin, O1; Sin, B1; Mungan, D1; Demirel, Y1;
Pinar, M2; Misirligil, Z1
1
Ankara University, School of Medicine, Department of
Immunolgy and Allergy, Ankara, Turkey; 2Ankara
University, Faculty of Science, Department of Biology,
Ankara, Turkey
Aim: Pre-seasonal allergen specic immu-
notherapy (SIT) has been shown to be
effective and safe in short-term. We wanted
to assess whether this immunotherapy
model has additional benet regarding
quality of life (QOL) or not.
Methods: Seventy-eight adult patients with
AR/asthma who live in Ankara city were
included in this prospective, controlled
observational study. After a follow-up per-
iod within 2007 pollen season, patients
were randomly selected for SIT (n:36) and
control groups (n:42) at 2008. Therapy
comprised two consecutive years presea-
sonal short-course of subcutaneous injec-
tions using a grasss pollen allergoid at
weekly intervals. The patients lled Turk-
ish version of Junipers rhinitis (RQoL) and
asthma (AQoL) quality of life question-
naires before and after each season. The
study groups were followed with symptom/
medication scores and evaluated at three
control visits. Both groups were allowed
for drug treatment as needed during the
pollen season. The number of gramineaea
pollens was counted daily in Ankara atmo-
sphere at the same periods.
Results: Twenty-seven out of 36 patients in
SIT Group and 27 out of 42 patients in
Control Group were re-evaluated at the
end of the second year of the study. The
signicant improvement in the mean score
of RQoL persisted at the second year of
SIT in comparison with the follow-up per-
iod (P = 0.000). Rhinitis symptom scores
were signicantly improved in patients
treated with SIT after the second year
(P = 0.001). In contrast, the AQoL scores
were tended more better than the rst year
of treatment (P = 0.049). There was a sig-
nicant decrease in the use of drugs
between the years in the SIT Group
(P = 0.000). However, control group did
not show changes in these parameters. No
side effect was seen in the study groups.
Conclusion: This study indicates that short-
term SIT is a good alternative in the man-
agement of pollinosis. At the second year,
quality of life for rhinitis and asthma
seemed to be still better with SIT as well
as reduced drug usage. However, we sug-
gest that persistence of clinical effectiveness
and the quality of life gain need boosting
injections for following years.
1683
Safety and compliance of cluster-immu-
notherapy achieving the maintenance
dose on the first treatment day with
highly polymerised allergen extracts
Dichmann, R1; Buscher, G2; Reuber, A2; Thieme, U3
1
Medical Practice, Pulmonology, Allergology, Witten,
Germany; 2Medical Practice, ENT, Essen, Germany;
3
Medical Practice, ENT, Duisburg, Germany
Background: Cluster immunotherapy with
a very short build up phase is convenient
for patients and physicians because of sav-
ing time consuming visits and injections.
Therefore cluster-immunotherapy has more
and more become a modern alternative to
conventional long lasting up-dosing sched-
ules. The safety prole and the patients
compliance of cluster-immunotherapy
achieving the maintenance dose on the rst
treatment day within 15 min has been ret-
rospectively evaluated in patients suffering
from allergic rhinitis/rhinoconjunctivitis.
Methods: For 57 patients (age 670 years)
suffering from IgE-mediated allergic rhini-
tis, conjunctivitis and/or mild allergic
asthma the safety data for up dosing on
the rst treatment day were retrospectively
633

analyzed. The patients had been treated
with specic subcutaneous immunotherapy
with different highly polymerized allergen
extracts (CLUSTOID) of house dust mites
(17 patients), grass pollen (19 patients) or
tree pollen (21 patients). The build up
schedule consisted of two injections on the
rst treatment day (0.2 ml and 0.5 ml with
an interval of 15 min) from one vial
(10 000 TU/ml). For documentation of the
safety prole any side effects following
these injections were evaluated for each
patient. For perennial immunotherapy one
injection (0.5 ml; 10 000 TU/ml) per month
has been given thereafter.
Results: The up dosing schedule with two
injections on the rst treatment day was
well tolerated by all 57 patients and no
local (grade1) or systemic side effects had
been observed. The patients compliance
was good; all patients were satised with
the fast up-dosing because of saving time
and injections. All patients continued the
treatment.
Conclusion: The fast up-dosing schedule
used for specic subcutaneous immuno-
therapy with different highly polymerized
allergen extracts achieving the maintenance
dose on the rst treatment day is safe and
results in good compliance of the patients.
1684
Quality of life in patients during allergy
immunotherapy
Gocki, J1; Knop, W2; Gawronska-Ukleja, E1; Bartuzi, Z1
1
Univ. Hospital 2, Collegium Medicum in Bydgoszcz,
University of Nicolaus Copernicus in Torun, Allergy,
Clinical Immunology and Internal Diseases, Bydgoszcz,
Poland; 2Univ. Hospital 2, Emergency Ward, Bydgoszcz,
Poland
Background: Allergy immunotherapy is
method of treatment allergy diseases. The
aim of this treatment is induce tolerance of
allergens. During treatment patient receive
increasing doses of allergens. When
patients achieve the maximal therapeutic
dose, this those should be repeated every
46 weeks trough 35 years. In classic
allergy immunotherapy the solution of
allergen is given in subcutaneous injections.
Regular injections is very important for
this therapy. Because possibility of anaphy-
lactic reaction this treatment should be
provided in specialized outpatient clinic.
This strict procedure of treatment could
have inuence on quality of life patient
during allergy immunotherapy.
Aim: The aim of this study was assessment
of quality of life patients during allergy
immunotherapy.
Methods: In the study take a part 100
patients with allergic diseases, at the age
from 18 to 51 years (36.8 year), 54 women
and 46 man. For assessment quality of life
634
Poster Session 73 Patient recorded outcomes and other paraclinical aspects in immunology
we use quality of life questionnaire WHO-
QOL-BREF. We assess quality of life in
four domains: somatic, psychological,
social and environmental. For more we
assess individual personal perception of
quality of life and individual personal per-
ception of health.
Results: Quality of life (QOL) in somatic
domain was very good in 49% of patients,
good in 41%, and in 10% of patients was
low. QOL in psychologic domain was very
good in 37% of patients, good in 51%,
and low in 12% of patients. QOL in social
domain was very god in 59% of patients,
good in 29%, and low in 12% of patients.
QOL in environmental domain was very
good 24%, good in 59%, and low in 17%.
Individual personal perception of quality
of life assess as very good 11% of patients,
good 80%, medium 7%, and bad 2% of
patients. Individual personal perception of
health assess as very good 2% of patients,
good 67%, medium 9%, and bad 22% of
patients.
Conclusions: Despite strict procedure of
treatment during allergy immunotherapy
most of patients has good quality of life in
all four domains. Individual perception of
quality of life and health most of patients
assessed as good.
1685
Motivating factors for the initiation of
allergen immunotherapy in severe, grass-
pollen-induced rhinoconjunctivitis
1 2 3
David, M ; Hut, M ; Meyer, M ; Capecce, M ;4
Polderman, C1
1
Stallergenes SA, Antony, France; 2Stallergenes SA,
The Netherlands; 3Stallergenes SA, Austria;
4
Stallergenes SA, Italy
Background: Subcutaneous injections and
sublingual drop and tablet formulations
are available for allergen immunotherapy
(AIT) in the treatment of severe, grass-pol-
len-induced rhinoconjunctivitis.
Methods: A study performed in Austria
(AT), Italy (I) and the Netherlands (NL)
in 2010 assessed AIT prescription habits in
specialist physicians and their patients suf-
fering from severe grass pollen-induced rhi-
noconjunctivitis (whether treated with AIT
or not).
Results: A total of 223 specialist physicians
(AT: 54; I: 113; NL: 56) and 716 adult
patients (AT: 190; I: 302; NL: 224) were
included. The specialists and the non-AIT-
treated patients generally considered that a
tablet formulation was easier to use and
could improve adherence. Between 67 and
79% of the specialists and between 59 and
75% of non-AIT-treated patients consid-
ered that a tablet formulation might moti-
vate patients who had previously been
reluctant to start AIT. Between 57 and
Allergy 66, Suppl. 94 (2011): 482642  2011 John Wiley & Sons A/S
75% of the AIT-treated patients stated
that the tablet formulation improved the
treatments acceptability. The physicians
and patients considered that the option of
a seasonal treatment regimen (about
6 months per year, versus. year-round
treatment) increased motivation, adherence
and satisfaction. A seasonal protocol
would encourage between 72 to 86% of
non-AIT patients to adopt this type of
treatment. Over 60% of the patients as a
whole considered that a seasonal protocol
is better suited to their disease state. This
opinion was shared by 75% of the physi-
cians in I but less than 50% of the physi-
cians in AT and NL. Between 70% (AT
and NL) and 80% (I) of the physicians
agreed that patients preferred seasonal
treatments, whereas the proportions of
treatments actually prescribed with a sea-
sonal regimen were only 35% (NL), 19%
(AT) and 62% (I).
Conclusion: According to specialist physi-
cians and their patients suffering from
severe rhinoconjunctivitis, tablet formula-
tions and seasonal protocols are signicant
motivating factors for the initiation of
AIT.
1686
Allergen immunotherapy regimens and
adherence in patients suffering from
severe grass-pollen-induced rhinocon-
junctivitis
David, M1; Hut, M2; Meyer, M3; Capecce, M4;
Polderman, C1
1
Stallergenes SA, Antony, France; 2Stallergenes SA,
The Netherlands; 3Stallergenes SA, Austria;
4
Stallergenes SA, Italy
Context: Allergen immunotherapy (AIT)
products (administered as subcutaneous
injection or as sublingual drops or tablets)
are available for the treatment of severe,
grass-pollen-induced rhinoconjunctivitis.
Methods: In 2010, a study assessing the
prescription habits of allergy specialist and
the preferences of patients with grass pol-
len allergy (whether treated with AIT or
not) was performed in Austria (AT), Italy
(I) and the Netherlands (NL).
Results: In total, 223 specialist physicians
(AT: 54; I: 113; NL: 56) and 716 adult
patients (AT: 190; I: 302; NL: 224) were
included. The AIT-treated patients gener-
ally consulted a physician (mainly allergol-
ogists in I and general practitioners in AT
and NL) 2 to 4 times a year, 1 to 3 years
after the rst appearance of their symp-
toms. Between 38 and 49% of the patients
requested a particular AIT treatment, these
were generally sublingually administered
products, chosen for ease and comfort of
use. Between 31% and 62% of patients
appeared not to have correctly understood
Poster Session 73 Patient recorded outcomes and other paraclinical aspects in immunology
the overall prescribed duration of the
course of AIT at the time of treatment
initiation, whereas over 86% of the physi-
cians said that they had clearly presented
it. In AT and NL, patients taking sublin-
gual AIT products stated that they were
treatment-compliant for an average of 4 to
7 months a year, whereas specialist physi-
cians had prescribed a year-round adminis-
tration regimen in more than 80% of
cases. In IT and NL, few patients whose
symptoms were insufciently controlled by
symptomatic medications had been offered
AIT (I: 14%; NL: 9% vs. AT: 48%).
When patients non controlled by sympto-
matic treatments refused AIT, in 57 to
88% it had been proposed as subcutaneous
injections.
Conclusion: Pre- and co-seasonally admi-
nistered sublingual AIT best meet the
expectations of patients with grass-pollen-
induced allergic rhinoconjunctivitis.
Allergy 66, Suppl. 94 (2011): 482642  2011 John Wiley & Sons A/S
1687
Immunotherapy in combination with pal-
liative surgery can lead to prolonged
remission of the disease in incurable
patients with basal cell carcinoma clini-
cal observation
Dorofeev, A1; Basova, L2; Buryakova, T3
1
Municipal Hospital of Elektrogorsk, Oncology,
Elektrogorsk, Russian Federation; 2Municipal Hospital
of Elektrogorsk, Dermatology, Elektrogorsk, Russian
Federation; 3Municipal Hospital of Elektrogorsk,
Ophtalmology, Elektrogorsk, Russian Federation
We conducted a combined treatment of
one patient with an incurable (at the stage
of development of modern medical sci-
ence), basal cell carcinoma of the lower
eyelid on the right with a broad and deep
spread of the soft tissues of the orbit, con-
junctiva and sclera of the eyeball (medical
history 4 years). Treatment was per-
formed with a palliative, but possible to
achieve remission at 9 months (at the time
of this writing).
Patient K., 72-years-old, refused radical
treatment at leading medical institutions in
the Moscow region, due to the prevalence
of tumor and the presence of severe com-
orbidity. When you try to palliative radio-
therapy, the patient developed temporary
cerebral ischemia, so this treatment was
interrupted after 1st session. We carried
out the following palliative treatment
(under local inltration anesthesia): (1)
spray-coagulation of the surface of the
tumor (with the exception of the conjunc-
tiva and cornea) in ve stages, held every
other day, and (2) the closure ap of tissue
defect of the upper eyelid (with excision of
the lacrimal glands) in the 14 days of start-
ing treatment, with enucleation of the eye-
ball was not carried out (technical
difculties), it remains sewn inside the
wound. Factor to warrant such crippling
manipulation was preoperative (initial)
total lack of vision in the affected eye due
to the spread of tumors in the tissue of the
cornea. At the same time were treated by
methylglucamine acridonacetati (cyclofer-
on) low-molecular weight inducer of
interferon (10 injections into surrounding
tumor tissue through day total dose
2500 mg). The wound is completely healed
at day 14 after surgery. Within 9 months
of follow-up revealed no recurrence of the
tumor. Previously, our colleagues reported
that immunotherapy is effective in treating
basal cell carcinoma of skin in the adju-
vant setting mode, in addition to operating
a radical way (including his special kind of
having a pool of squamous cells metatyp-
ical skin cancer). Described by our case
suggests that the indications for immuno-
therapy can be extended and applied in
cases of inoperable tumors of the skin in
addition to palliative surgery with the hope
of a complete radical cure of this tumor or
the achievement of long-term remission.
This thesis requires further research in this
direction.
635
 Poster Session 74

Allergy care and education

Results: Files: drug allergy (30% older than 60 years) -

1688
Prevalence of allergic diseases There is no signicant difference in the
Demographics of patients treated in an
Prevalence of allergic diseases by age prevalence of allergic diseases in different
allergy unit in a year
and sex populations depending on the area.
Alba, P; Calderon, R; Iglesias, I Allergic disease prevalence populations
Hospital Manises, Allergy, Valencia, Spain Table 1. For abstract 1668.
Conclusion: The most frequent reasons for
consultation were: respiratory allergy (rhi- Prevalence of allergic diseases Total %
Background: Try to make a descriptive
nitis), which represents 55% of visits to 2931
study of patients attending an allergy ser-
our service. Asthma to 23.6%, urticaria in Rhinitis (R) 1615 55.1
vice at a privately run public hospital dur-
13% and almost 9.5% of drug allergy -The Conjunctivitis (C) 1185 40.4
ing its rst year of operation, in order to
60.8% of patients were women. Among Asthma (A) 691 23.6
provide important evidence for further
patients attending our service a higher pro- Angioedema 136 4.6
study and improve the quality of service Anaphylaxis 54 1.8
portion of women with drug allergy, hives,
offered to patients. Urticaria 386 13.2
dermatitis and ODS than in men. -For
Methods: We collected demographic and Dermatitis 223 7.6
ages are objectied: *Most patients treated
clinical data of all rst visits (2931), from an Allergy drugs 278 9.5
between ages 20 and 50 years, with a distri-
area of 196 379 inhabitants, arriving at SAO 72 2.5
bution of the prevalence of allergic diseases R+C 1163 39.6
allergy unit of Manises Hospital from May
similar to that of all patients logically. *In R+A 590 20
2009 until May 2010, being attended by two
children under 20 years are also prevalent R+C+A 476 16
physicians. We performed a statistical study
rhinitis, but asthma occur more frequently Allergic respiratory disease 1735 59
showing types of allergic diseases in relation
(nearly 50% under 10 years) *In over
to various demographic characteristics that
50 years there is an increasing suspicion of
were used SPSS 15.0 statistical package.

Table 1. For abstract 1688b.

Total Men Women 09 years 1019 years 2029 years 3039 years 4049 years 5059 years 6069 years >70 years
Men 68 178 256 283 161 97 67 28
Women 78 193 360 406 300 220 147 89
Total 2931 1148 1783 146 371 616 689 461 317 214 117
Rhinitis 1615 680 935 79 247 419 421 251 120 62 16
Conjunctivitis 1185 483 702 40 172 330 308 191 89 44 11
Asthma 691 268 423 62 127 148 172 97 51 27 7
R+C 1163 474 689 39 167 316 304 186 85 88 11
R+A 590 238 352 47 114 133 152 87 33 20 4
R+C+A 475 184 291 27 89 113 119 74 29 20 4
Angioedema 136 49 87 6 16 25 42 19 12 13 3
Anaphylaxis 54 22 32 0 5 14 18 7 6 4 0
Urticaria 386 135 251 16 41 84 87 66 45 30 17
Dermatitis 223 70 153 17 19 37 54 30 36 23 7
Drug allergy 278 88 190 4 10 26 41 55 46 54 42
SAO 72 24 48 3 5 21 19 11 6 6 1

Table 1. For abstract 1688c.

Prevalence of allergic diseases Aldaya Bunol Cheste Chiva Manises Mislata Quart Ribarroja Turis Valencia
TOTAL 388 139 145 189 582 227 494 350 116 115
Rhinitis (R) 226 (58%) 85 (61%) 70 (48%) 110 (58%) 341 (59%) 84 (37%) 283 (57%) 201 (57%) 60 (52%) 66 (57%)
Conjunctivitis (C) 168 (43%) 68 (49%) 49 (34%) 83 (44%) 242 (42%) 66 (29%) 194 (39%) 151 (43%) 47 (40%) 50 (43%)
Asthma (A) 102 (26%) 39 (28%) 33 (23%) 51 (27%) 140 (24%) 40 (18%) 124 (25%) 76 (22%) 22 (19%) 25 (22%)
Angioedema 18 (5%) 9 (6%) 5 (3%) 8 (4%) 21 (4%) 0 19 (4%) 24 (7%) 4 (3%) 12 (10%)
Anaphylaxis 7 (2%) 3 (2%) 3 (2%) 1 (0.5%) 19 (3%) 1 (0.4%) 4 (0.8%) 4 (1%) 2 (1.7%) 3 (2.6%)
Urticaria 56 (14%) 23 (16%) 22 (15%) 18 (9%) 84 (14%) 9 (4%) 62 (13%) 51 (15%) 13 (11%) 17 (15%)
Dermatitis 40 (10%) 10 (7%) 8 (5%) 16 (8%) 49 (8%) 7 (3%) 37 (7%) 29 (8%) 5 (4%) 6 (5%)
Drug allergy 38 (10%) 12 (8%) 8 (5%) 15 (8%) 62 (11%) 10 (4%) 51 (10%) 41 (12%) 10 (9%) 12 (10%)
SAO 13 (3%) 3 (2%) 2 (1%) 0 16 (3%) 3 (1%) 13 (3%) 7 (2%) 3 (2.7%) 5 (4%)
R+C 163 (40%) 65 (48%) 47 (33%) 83 (44%) 241 (41.5%) 66 (29%) 190 (38%) 148 (42%) 47 (40%) 49 (42%)
R+A 89 (21%) 36 (27%) 28 (21%) 44 (21%) 125 (22%) 33 (14.5%) 101 (19.5%) 67 (19.5%) 18 (15%) 19 (15%)
R+C+A 70 (18%) 31 (24%) 21 (14.5%) 35 (18%) 95 (17%) 26 (11%) 81 (15%) 58 (17%) 17 (14%) 16 (14%)

636 Allergy 66, Suppl. 94 (2011): 482642  2011 John Wiley & Sons A/S
Poster Session 74 Allergy care and education
1689
Tracking of patients with anaphylaxis
across the public health institutions of a
city in the metropolitan area of Madrid
region
Moro, M1; Tejedor, M1; Mugica, M1; Esteban, J2;
Gomez-Traseira, C1; Cardenas, R1; Rosado, A1; Vila, C1;
Sacristan, J3; Hernandez, A4; Acosta, M1
1
Hospital Universitario Fundacion Alcorcon, Allergy,
Alcorcon, Spain; 2Universidad Rey Juan Carlos,
Preventive Medicine and Public Health Department,
Alcorcon, Spain; 3Primary Care Health Center Gregorio
Maranon, Alcorcon, Spain; 4Primary Care Health Center
Granero Vicedo, Alcorcon, Spain
Background: Patients with anaphylaxis
were tracked across the different public
health institutions in the city of Alcorcon
(Spain).
Methods: Episodes of anaphylaxis were
recovered using validated alphanumeric
strings in different elds of electronic clini-
cal records used in the different public clin-
ical settings in the city of Alcorcon
(Primary Care, Emergency Department,
hospitalized patients and Allergy OutPa-
tient Clinic). This strategy show 91.7% of
sensitivity with respect to a comprehensive
revision of all emergency episodes from
March and April 2004. The period of study
was between 1 January 2004 and 31
December 2005. Anaphylaxis was dened
using the NIAID-FAAN criteria: involve-
ment of at least two organs or systems, or
presence of hypotension alone as the only
manifestation after exposure to a known
allergen. In order not to duplicate episodes
of anaphylaxis from different databases,
we assessed the presence of similar or iden-
tical organ or system involvement and the
same cause. Two or more anaphylaxis epi-
sodes in the same patient with a similar
clinical prole and the same cause recorded
in two or more places were considered the
same episode if the interval between them
was less than 3 months.
Table 1. For abstract 1689.
Paths followed by patients with anaphylaxis episodes
Primary care and Allergy Outpatient Clinic
Emergency Department. Primary care and Allergy
Outpatient Clinic
Emergency Department only
Emergency Department and
Allergy Outpatient Clinic
Emergency Department and primary care
Allergy Outpatient Clinic only
Primary care only
Hospitalized patients. Primary care and Allergy
Outpatient Clinic
Emergency Department. Hospitalized patients.
Primary care and Allergy Outpatient Clinic
Hospitalized patients and primary care
Only hospitalized patients
Hospitalized patients and Allergy Outpatient Clinic
Emergency Department and hospitalized patients
Allergy 66, Suppl. 94 (2011): 482642  2011 John Wiley & Sons A/S
Results: There were 336 episodes overall. Results: During ragweed seasons, allergic
The interquartile range for age was 19.11 patients reported higher levels of general
51.46 years. Comparison of the different fatigue and mental fatigue, but not physi-
care levels revealed that 75.6% of the cal fatigue, as well as reduced motivation.
patients were assessed at the Allergy Out- Patients described experiencing feelings of
patient Clinic, 71.43% at a primary care greater sadness and reduced pleasurable
center, and 55.95% at the Emergency engagement. Increased anxiety or emo-
Department (Table 1). Patients in our ser- tional distress was not reported.
ies were generally attended in two or more Conclusion: These ndings suggest that
places (76.78%), with Allergy Outpatient having allergic reactions to ragweed pollen
Clinic and a primary care center being the causes signicant fatigue and mood
most typical combination (58.9%). Only changes in at least a subgroup of patients.
23.22% of patients were attended at a sin- Psychoneuroimmunology and medical
gle center, with a single visit to Emergency genetics research suggests that allergic reac-
Department being the most common tions engender biochemical changes that
(11.31%) (Table 1); 3.57% of cases were directly affect the central nervous system.
attended at the Allergy Outpatient Clinic
only and 2.68% at a primary care center
only.
1691
Conclusion: Patients in our series were gen-
Levels of evidence for physiotherapy
erally attended in two or more places interventions in asthma. a critical review
(76.78%), with Allergy Outpatient Clinic of clinical practice guidelines
and a primary care center being the most
Nowobilski, R1; W3och, T2; P3aszewski, M3; Szczeklik, A4
typical combination (58.9%). Only 23.22% 1
Jagiellonian University School of Medicine, Cracow,
of patients were attended at a single center. Poland, Department of Medicine, Cracow, Poland;
2
University School of Physical Education, Cracow,
Department of Rehabilitation, Poland; 3University
School of Physical Education, Warsaw, Faculty of
1690 Physical Education in Biala Podlaska, Poland;
4
Jagiellonian University School of Medicine, Cracow,
Effects of seasonal allergic rhinitis on Department of Medicine, Poland
fatigue levels and mood
Adwan, Z Background: Based on a growing body of
SMS Hospital, Beirut, Lebanon scientic evidence, pulmonary rehabilita-
tion has emerged as a recommended stan-
Objective: Many allergy patients complain dard of care for patients with chronic lung
of fatigue, moodiness, and dysphoria dur- diseases. Nonetheless, despite being listed
ing their allergy seasons. This study evalu- in different clinical practice guidelines,
ated the effect of symptomatic allergic multiplicity and variety of physiotherapy
rhinitis on both fatigue level and mood. methods and techniques recommended for
Methods: Symptomatic ragweed allergic patients with bronchial asthma require
rhinitis patients on no medications and careful assessment of validity and reliabil-
healthy control subjects completed the ity of evidence in this subject.
Multi-Dimensional Fatigue Inventory and Methods: The American Association of
the Positive Affect-Negative Affect mood Cardiovascular and Pulmonary Rehabilita-
rating scales in an in-out-in ragweed season tion (AACVPR, 2010), the British Tho-
research design. racic Society and Association of Chartered
Physiotherapists in Respiratory Care
(BTC/AC.PRC, 2009) and the Global
Strategy for Asthma Management and Pre-
N % Cumulative % vention/ Global Initiative for Asthma
95 28.27 28.27 (GINA, updated 2008) clinical practice
90 26.79 55.06 guidelines were analyzed in the context of
recommendations of physiotherapy meth-
38 11.31 66.37 ods and techniques for patients with
28 8.33 74.7 asthma. Individual original studies,
extracted from subsequent guidelines, were
27 8.04 82.74 also analyzed. The Physiotherapy Evidence
26 7.74 90.48
Database (PEDro) and the Scottish Inter-
12 3.57 94.05
9 2.68 96.73
collegiate Guideline Network (SIGN)
scales were used to assess the quality of
4 1.19 97.92 evidence from original studies, levels of
evidence and grades of recommendations,
2 0.6 98.52 respectively.
2 0.6 99.12 Results: Some papers were based on small
2 0.6 99.72 studies, limited to short-term outcomes
1 0.3 100 and with a single treatment session. More-
637

over, most studies presented single out-
come measures and crossover designs. Rec-
ommendations from clinical guidelines are
more than once inconsistent.
Conclusion: Further analyses of Health
Related Quality of Life (HRQoL), working
capacity, long-term outcomes and hospital
stay, are needed. The evidence from studies
in patients with chronic obstructive pulmo-
nary diseases and cystic brosis cannot be
directly extrapolated to asthma subjects.
Future research should focus on more
appropriate matching of the physiological
effects of individual physiotherapy tech-
niques to the pathophysiology of bronchial
asthma. Dosage and treatment methodol-
ogy ought to be standardized.
1692
Patient participation in lung research
agenda setting: methodology and value
Laan, D1; Elberse, J2; Teunissen, T1; Broerse, J2; de
Cock Buning, T2; de Boer, P1
1
Netherlands Asthma Foundation, Research,
Amersfoort, The Netherlands; 2Free University, Athena
Institute, Amsterdam, The Netherlands
Background: Patients in lung research are
often regarded as being study objects. As
active involvement of patients in research
seems useful, we previously developed a
novel method involving patients in lung
research policy and decision making. A
shortened model to update a research
agenda was not available.
Aim: Develop a brief method to update
the research agenda based on active patient
participation.
Methods: The update method included
three steps. (1) individual interviews and a
plenary discussion with lung scientists/doc-
tors (group SD; n = 20), resulting in a sci-
entic agenda.(2) focus group discussions
with patients with chronic lung diseases
(group P; n = 31). Data were validated
and prioritized using a web-based question-
naire in group P (n = 165) (3) grouping
and merging resulting data from both
groups into two different lists: an updated
list for asthma and COPD research, and a
new list for other chronic pulmonary dis-
Table 1. For abstract 1693.
Groups Parameter
Controlled (86) average
SD
Partly controlled (50) average
SD
Uncontrolled (76) average
SD
ANOVA test F 21.02
(Fkr = 3.04) p <0.0001
638
eases. Group SD provided feedback on
attainability.
Results: The update method involving all
groups required 5 months. Patients were
satised with this methodology and able to
provide their perspectives. Two different
workable agendas resulted. The overlap
between research priorities of all groups in
each agenda was huge regarding all
research areas. The overlap between both
agendas was also considerable. We noted
differences with respect to e.g. co-morbidi-
ties (P), smoking interventions (SD), and
disease-specic research themes.
Conclusion: Updating research priorities by
this model proved useful for eliciting
research priorities from scientists, doctors
and patients. Involving patient experiences
identied and exemplied a new research
focus on chronic lung diseases in The
Netherlands.
1693
Quality of life in asthma
Szynkiewicz, E1; Cegla, B2; Popow, A3; Filanowicz, M2;
Dowbor-Dzwonka, A3; Bartuzi, Z3
1
Collegium Medicum in Bydgoszcz, Nicolaus
Copernicus University in Torun, Internal Nursing, Clinic
of Allergology, Clinical Immunology and Internal
Diseases, Bydgoszcz, Poland; 2Collegium Medicum in
Bydgoszcz Nicolaus Copernicus University in Torun,
Internal nursing, Bydgoszcz, Poland; 3Poland
Background: Bronchial asthma affects peo-
ple all over the world. It is a disease with
variable course as well as a long-lasting
and incurable illness inuencing the
patients quality of life. In Poland it occurs
in more than 3 million people. Evaluation
of quality of life in patients with uncon-
trolled asthma of varying degrees of sever-
ity according to GINA 2006 denitions.
Methods: The study using the method of
diagnostic poll was conducted among 212
patients of the clinic and allergy out-
patient clinic in 20082010. Patients were
divided into three groups. The criterion of
the division was the degree of asthma con-
trol: (Gina 2006 denitions) controlled (86
patients), partially-controlled (50) and
uncontrolled (76). To determine the quality
of life, the Polish version of WHOQOL-
Physical Psychological Level of
domain domain independence
0.639 0.607 0.676
0.162 0.130 0.147
0.526 0.602 0.557
0.146 0.131 0.171
0.485 0.595 0.485
0.154 0.127 0.160
0.15 30.23
0.86 <0.0001
Allergy 66, Suppl. 94 (2011): 482642  2011 John Wiley & Sons A/S
Poster Session 74 Allergy care and education
100 scale was used. It contains 100 ques-
tions falling within the six life domains:
physical, psychological, independence,
social, environmental and spirituality. The
study used ANOVA test and Tukey RIR
test.
Results: RIR Tukey test found that the
level of quality of life in the physical
domain is signicantly higher in patients
with controlled asthma than in other
groups. Levels of quality of life in the
groups with partially controlled and
uncontrolled asthma do not differ from
each other. In the level of independence
domain, signicant differences in quality of
life were found in all compared groups.
Conclusion: (1) Based on the conducted
research, it was found that there are signi-
cant differences in HRQL in patients,
depending on the degree of asthma con-
trol.
(2) Patients diagnosed with uncontrolled
or partially controlled asthma show much
lower quality of life in the physical domain
and level of independence.
(3) Patients with bronchial asthma show
no differences in other domains (psycho-
logical, social relationships, environmental
and spirituality).
1694
Self-treatment of patients with recurrent
anaphylaxis episodes
Mugica Garcia, M; Tejedor Alonso, M; Rojas Perez-
Ezquerra, P; Moro Moro, M; Rosado Ingelmo, A; Vila
Albelda, C; Almanzar Abreu, W; Acosta Rivera, M
Hospital Universitario Fundacion Alcorcon, Allergy
Unit, Madrid, Spain
Background: Information on the self-man-
agement of patients with recurrent episodes
of anaphylaxis is scarce. We analyzed self-
treatment with an epinephrine auto-injector
and oral medication (H1-antihistamines
and corticosteroids) in a cohort of patients
who experienced recurrent anaphylaxis.
Methods: The cohort comprised patients
with episodes of anaphylaxis recorded on
the allergic diseases database of the Allergy
Unit of Hospital Universitario Fundacion
Alcorcon (Madrid.Spain) between 1998
Social Environment Spirituality
relationships domain domain
0.705 0.625 0.412
0.133 0.091 0.211
0.692 0.612 0.524
0.154 0.095 0.226
0.689 0.610 0.546
0.127 0.097 0.221
0.31 0.63 0.51
0.74 0.53 0.60
Poster Session 74 Allergy care and education
and 2005. A questionnaire was sent to all
patients in the cohort (1590) and three
phone calls were made to non-responders.
The questionnaire recorded information on
new episodes of anaphylaxis, their clinical
and demographic characteristics, and the
mode of self-treatment administered. Ana-
phylaxis was dened using the NIAID-
FAAN criteria: involvement of at least two
organs or systems, or presence of hypoten-
sion alone as the only manifestation after
exposure to a known allergen. Recurrence
was dened as a new episode in which the
generic cause was the same as in the previ-
ous episode.
Results: Overall, 34.84% of patients expe-
rienced recurrence of anaphylaxis. With
regard to the epinephrine auto-injector,
37.46% of patients reported that they had
not received any prescription after their
allergy assessment, 34.46% bought the
device, and 30.74% used it. Acquisition
(P = 0.013) and use (P = 0.014) of an
epinephrine auto injector was signicantly
different between subtypes of anaphylaxis.
With regard to oral medication, 30.85%
patients reported that they had not
received any prescription after their allergy
assessment, and 54.21% reported taking
medication. No signicant differences were
observed between subtypes of anaphylaxis
in relation to oral medication (P = 0.35).
Similarly, no differences in the percentage
of acquisition of oral medication were
observed in any of the subtypes
(P = 0.072). Comparison of both types of
treatment revealed that, in all subtypes of
anaphylaxis, the acquisition of oral medi-
cation was always higher than that of the
epinephrine auto-injector and that taking
oral medication was more common than
self-treatment with the auto-injector.
Conclusions: We observed that less than
half of our cohort acquired and used the
epinephrine auto-injector after their allergy
assessment. In all subtypes of anaphy-
laxis, acquisition and administration of
oral medication was more common than
acquisition and self-administration of the
epinephrine auto-injector.
Table 1. For abstract 1694.
Epinephrine Oral
auto-injector medication
acquisition acquisition
Anaphylaxis % %
subtypes
Food 77.36 88.75
Drugs 42.31 69.23
Exercise 50 100
Anisakis 33.33 77.27
Hymenoptera 85.71 88.89
Idiopathic 70 87.5
P 0.013 0.072
Allergy 66, Suppl. 94 (2011): 482642  2011 John Wiley & Sons A/S
1695
Epidemiology of anaphylaxis in Pie-
monte, Italy: data from the regional reg-
istry of severe allergic reactions
Raie, A1; Mietta, S2; Nebiolo, F3; Cadario, G4;
Galimberti, M5; Heffler, E6; Rolla, G7
1
University of Torino, Ospedale Mauriziano Umberto I,
Allergy and Clinical Immunology, Center for Severe
Allergic Reactions of Piemonte Region, Torino, Italy;
2 Conclusion: The high prevalence of hyme-
University of Torino, Ospedale Mauriziano Umberto I,
Allergy and Clinical Immunology, Torino, Italy;
3
Mauriziano Hospital, Umberto IA, Allergy and Clinical
Immunology, Turin, Italy; 4Molinette Hospital, Allergy
and Clinical Immunology, Center for Severe Allergic
Reactions of Piemonte Region, Turin, Italy; 5Ospedale
Maggiore e della Carita, Allergy and Clinical
Immunology, Novara, Italy; 6University of Torino,
Allergy and Clinical Immunology, Torino, Italy;
7
University of Torino, Ospedale Mauriziano Umberto I,
Allergy and Clinical Immunology, Turin, Italy
Background: The incidence of severe aller-
gic reactions in general population is lar-
gely unknown and information about
triggering factors is lacking.
Methods: We reviewed anaphylaxis cases
from the database of the Piemonte Region
Reference Center for Severe Allergic Reac-
tions. The registry monitors a population
of 4 400 000 inhabitants, with real-time
registration of regional cases of severe
reactions, and collects data mandatory for
prescribing self-injectable epinephrine reim-
bursed by Regional Health System. Ana-
phylaxis cases were evaluated according to
NIAID/FAAN diagnostic criteria, and
assigned to one in three levels of probabil-
ity of anaphylaxis using a clinical checklist
based on recommendations of the Brighton
Collaboration for anaphylaxis following
immunization.
Results: Among the 2461 reported cases of
severe allergic reactions, 1434 (58.3%
[Females 44.3%, Males 55.7%, mean age
36.8 years (range 087 years)] were classi-
ed as anaphylaxis. The causes of anaphy-
laxis were: hymenoptera venom in 47.8%
of cases (18.5% in subjects <18 years ver-
sus 55.1% in subjects 18 years,
P < 0.0001), foods in 39.7% (76.9% ver-
sus 30.4%, P < 0.0001), drugs in 7.7%
(2.4% versus 9.1%, P < 0.0001), other
causes in 1.9%, idiopathic in 2.9%. Skin,
respiratory and gastrointestinal symptoms
Taking
Self-treatment oral
with epinephrine medication
% %
34.92 80.77
59.57 71.43
0 40
78.57 85
44.44 87.5
50 80
0.014 0.35
were more frequent in children compared
with adults (95.8% versus 89.4%,
P = 0.001; 79.4% versus 73.2%, P =
0.034; 38.1% versus 23.8%, P < 0.0001,
respectively), while cardiovascular involve-
ment was more frequent in adults than in
children (55.1% versus 22.7%, P <
0.0001).
noptera compared to drug-induced ana-
phylaxis we observed is probably explained
by the required prescription procedure of
self-injecting epinephrine. Differences in
organ involvement and triggers of anaphy-
laxis were found between young and adult
patients. We believe that a unied data col-
lection system could help to understand
better the occurrence and circumstances of
severe allergic reactions.
1696
InterAll an allergy electronic health
record supporting the molecular allergol-
ogist in the daily activities: extracting
and visualising real time statistical data
Santoro, M1; Rasi, C1; Monti, S1; Nuti, S2; Scala, E1;
Zennaro, D1; Palazzo, P1; Liso, M1; Alessandri, C1;
Carabella, G3; Mari, A1
1
IDI-IRCCS, Center for Molecular Allergology, Rome,
Italy; 2Allergy Data Laboratories s.c., R&D, Latina, Italy;
3
Panservice s.n.c., R&D, Latina, Italy
Background: To describe a user-friendly
tool helping clinicians in allergy diagnosis
by displaying in a web browser up-to-date
summary statistics and charts.
Methods: At our Center there is a fully
computerized system for patients data
handling. This web-technology based sys-
tem is called InterAll (Allergy Data Labo-
ratories s.c., Italy), where around 56 812
patients demographics and clinical data
are stored (as of January 17, 2011). Data
from the ISAC system used for IgE detec-
tion (PMD, Austria) are stored in the same
database. InterAll has been created using a
MySql database and a PHP web interface.
Within the InterAll system we built an
automated statistical tool by using R-
CRAN, for analyzing data, generating
downloadable high quality, ready to pub-
lish charts, and the correlated datasets and
PHP for visualizing them in a web page.
An automated R-script brings up-to-date
data analyses once a day, producing charts
and graphs, and generating an up-to-date
PHP web page of statistics. All operations
are in an almost-real time way.
Results: InterAll-Statistics tool analyzes
patients distribution by sex, age (current
and at the time of registration in the DB)
and by age and sex combined. It gives g-
ures about how many clinical tests have
been performed and in how many patients,
it draws the absolute frequency distribution
639

of how many patients performed n tests
(n ranging from one to the highest number
of clinical test in one patient). The tests
analytical procedure can go into a more
detailed analysis evaluating the number of
different tests, i.e. skin test, specic IgE
detected by singleplex systems (i.e. Immu-
noCap, Immulite) or by multiplex systems
(i.e. ISAC103, 89, 85, 81, 79, 76, and 74)
and whether they are clinical routine tests
or experimental ones. As far as the Inter-
All e-record allows also storing biological
samples related to patients, overall avail-
ability of sera is also given. Finally, the
tool draws the total and relative frequency
of patients positive to each of 103 different
allergens tested using e.g. ISAC103. Its
relevant to note that our tool is developed
using open source software and it is inde-
pendent from the specic data-set.
Conclusion: Herein we report the develop-
ment of a statistical tool helping clinicians
in visualizing and analyzing up-to-date
demographics and clinical features of the
examined allergic population. Further
dynamic data analysis can be developed
and integrated in the InterAll-Statistics
page.
1697
Training of trainers on epinephrine auto-
injector use
Arga, M1; Bakirtas, A1; Catal, F2; Derinoz, O3; Harmanci,
K4; Razi, C5; Ergocen, S6; Demirsoy, S1; Turktas, I1
1
Gazi University Hospital, Department of Pediatric
Allergy and Asthma, Ankara, Turkey; 2Fatih University
Hospital, Department of Pediatric Allergy and Asthma,
Ankara, Turkey; 3Gazi University Hospital, Department
of Pediatric Emergency, Ankara, Turkey; 4Ministry of
Health Ankara Diskapi Childrens Diseases Training and
Research Hospital, Department of Pediatric Allergy and
Asthma, Ankara, Turkey; 5Ministry of Health Ankara
Kecioren Training and Research Hospital, Department
of Pediatric Allergy and Asthma, Ankara, Turkey;
6
Ankara University Faculty of Medicine, Department of
Biostatistics and Medical Informatics, Ankara, Turkey
Background: The majority of physicians do
not know how to use epinephrine auto-
injectors. This displays that current educa-
tion of physicians on anaphylaxis is inade-
quate for a thorough practice. We
hypothesize that a theoretical lecture
together with a practical session on epi-
nephrine auto-injector use will improve its
proper use by physicians.
Methods: Residents, specialists and consul-
tants from General Pediatrics excluding
allergists and allergy fellows were included
in this study. All physicians were given an
eight item questionnaire followed by a
practical session scoring and timing ability
to use epinephrine auto-injector trainer.
This ensued with one to one hands-on
training on correct auto-injector use.
Finally, a joint theoretic lecture on ana-
phylaxis including re-demonstration of epi-
640
nephrine auto-injector use was given. All
physicians were scored a second time on
use of epinephrine auto-injector 6 months
later.
Results: One hundred fty one of 196 par-
ticipants completed all steps of the study in
four tertiary hospitals. Correct use of epi-
nephrine auto-injector improved from
23.3% to 74.2%, mean score from
3.49 1.14 to 4.66 0.65, need for pro-
spectus from 91.4% to 29.1% and mean
time to administer auto-injector from
28.01 6.22 s to 19.62 5.01 s
(P < 0.001 for each). The rate of most
common mistakes during auto-injector use
decreased but the ranking did not change.
Conclusion: An integrated theoretic and
practical education increased correct of
epinephrine auto-injector use by physi-
cians. Ongoing mistakes despite this educa-
tion may be related with its design.
1698
Dental health in allergic patients
Atayik, E1; Epozturk, K1; Bingol, A2; Abadoglu, O1
1
Cumhuriyet University, Faculty of Medicine,
Department of Immunology and Allergic Diseases,
Sivas, Turkey; 2Cumhuriyet University, Faculty of
Medicine, Department of Chest Diseases, Sivas, Turkey
Background: Some studies investigated the
prevalance of dental disorders in asthmatic
patients. However, scant data are available
considering the relationship between dental
health and other allergic diseases.
Methods: A questionnaire about the pres-
ence of allergic diseases, dental diseases,
any previos dental treatment, and tooth
brushing habits was applied face-to-face to
the patients in the outpatient clinic of
Allergic Diseases and to a control group
with no allergic diseases.
Results: The study involved 395 patients
(52% allergic rhinitis, 47% asthma, 14%
chronic urticaria, 13% drug allergy, and
8% allergic contact dermatitis) and 286
controls with similar tooth brushing habits.
Eighty-eight percent of the patients had
dental caries, a similar ratio was found in
the control group (P > 0.05). Further-
more, there were no differences between
asthmatic patients and those without
asthma, and between the patients who
receive high dose inhaled steroids and
those who do not (P > 0.05).
Conclusion: The most common dental
problem in allergic patients is dental caries.
Inhaled steroid demonstrated no adverse
effect on dental health. Any patient should
be instructed for regular tooth brushing.
Allergy 66, Suppl. 94 (2011): 482642  2011 John Wiley & Sons A/S
Poster Session 74 Allergy care and education
1699
Make up for the epinephrine auto-injec-
tor: effect on its use by untrained users
Bakirtas, A1; Arga, M1; Catal, F1; Derinoz, O2; Demirsoy,
S1; Turktas, I1
1
Gazi University Hospital, Department of Pediatric
Allergy and Asthma, Ankara, Turkey; 2Gazi University
Hospital, Department of Pediatric Emergency, Ankara,
Turkey
Background: Part of the problems related
with proper use of epinephrine auto-injec-
tor may be related with the design of the
auto-injector itself. To investigate whether
minor modications in the design of cur-
rently available epinephrine auto-injector
ease its use and abrogate common use
errors.
Methods: All interns except the ones who
previously worked in allergy department in
a medical school were invited to the study.
Two identical epinephrine auto-injector

trainers (Epipen trainer ) were used, one of
which was modied by changing gray
safety cap to red and placing a yellow
arrow pointing to the black injection tip.
Written and visual instruction sheet for
each trainer was provided. Participants
were asked to demonstrate the use of Epi-
pen trainer either with the original or the
modied one. They were scored and timed
for demonstration.
Results: One hundred sixty-four interns
participated in the study among 224 invited
(73.2%). The number of participants cor-
rectly demonstrating the use of epinephrine
auto-injectors was 22.6% in original and
65% in modied trainer groups (P < 0.001)
Mean time to administer trainers were
26.78 10.6 s and 15.88 2.55 s; and
total median scores were 3.08 1.48 and
4.47 0.84 in original and modied
groups, respectively (P < 0.001 for both).
Signicantly fewer participants tackled with
black tip of trainer in order to eject needle
after removing safety cap in modied (5%)
compared to original trainer group (45.2%)
(P < 0.001).
Conclusion: Few and simple modications
in the design of commonly available epi-
nephrine auto-injector were found effective
in increasing its correct use and decreasing
common use errors by untrained users.
1700
Complementary and alternative medicine
in children with allergic diseases
Guner, S; Kilic, M; Ozturk, F; Yildiran, A; Sancak, R
Ondokuz Mayis University, Department of Pediatric
Allergy and Immunology, Samsun, Turkey
Background: The popularity of comple-
mentary and alternative medicine (CAM),
especially for chronic conditions, is rising,
but there are limited data about the
Poster Session 74 Allergy care and education
frequency of use for allergic children in
our region. This study aimed to determine
the prevalence of CAM use, and to identify
factors associated with their use in the chil-
dren and for those with asthma, allergic
rhinitis and eczema.
Methods: A cross-sectional survey of chil-
dren (118 years) who presented to the
Department of Pediatric Allergy of Ond-
okuz Mayys University Hospital (Samsun,
Turkey) between JanuaryDecember 2010
was conducted. Data were collected by
questionnaire that included 19 questions.
After short information, parents completed
a questionnaire. Later, all answers were
checked for missing or unperceived ques-
tions.
Results: A total of 506 (70.6%) with
asthma, 155 (21.6%) with allergic rhinitis,
and 56 (7.8%) with eczema, total 717 chil-
dren were included to the study. The mean
the age of 331 (46.2%) girls were
7.4 4.0 years and 386 (53.8%) boys were
6.7 3.9 years. The prevalence of the par-
ents information about at least one of
CAM was 88.6% but the prevalence of
CAM use among children with allergic dis-
eases was 39.6%. The most popular forms
of CAM were herbal medicine (56.4%),
Turkish wild honey (54.1%), quail eggs
(23.9%), sh oil (11.6%), vitamins (7.8%),
and royal jelly (6.4%). Age, sex, geo-
graphic region, mothers education levels,
type and control level of allergic diseases
were insignicant between the groups that
used and did not use CAM. But father
educational level and families incomes were
signicantly high in the CAM group
(P = 0.03 and P = 0.006, respectively).
Conclusion: A higher level of education
and socio-cultural preference is more com-
mon in the CAM group. Contrary to the
expectations, the type and control level of
allergic diseases did not directly affect the
use of CAM.
1701
Turkish physicians dealing with allergic
diseases: their knowledge about comple-
mentary and alternative medicine, and
attitudes towards complementary and
alternative medicine
Arguder, E1; Yilmaz, I2; Ates, C3; Misirligil, Z2;
Bavbek, S2
1 tations are admitted to the Emergency
Ankara Ataturk Training and Research Hospital,
Department of Chest Diseases, Ankara, Turkey; 2Ankara
University School of Medicine, Department of Allergy
and Immunology, Ankara, Turkey; 3Ankara University
School of Medicine, Department of Biostatistics,
Ankara, Turkey
Background: Allergic disorders; including
rhinitis, asthma, and atopic dermatitis are
some of the most common chronic disor-
ders which are associated with increased
CAM use. However, there is no study pub-
Allergy 66, Suppl. 94 (2011): 482642  2011 John Wiley & Sons A/S
lished reporting physicians attitudes
towards CAM dealing with allergic dis-
eases. We aimed to evaluate knowledge
and attitude of the physicians who deal
with allergic patients about CAM in this
study.
Methods: A structured questionnaire
related to respondents demographics,
knowledge about CAM and attitudes
toward CAM was prepared by the study
team and was applied to the physicians
who participated in Turkish National
Society of Allergy and Clinical Immunol-
ogy Congress-2009.
Results: Two hundred-forty two physicians
responded to the questionnaire. The median
(min-max) age of the respondents was 36.0
(26.064.0) years. More than half of them
were female (67.8%). Among all, 47.8% was
specialist, of whom 49.3% was paediatrician
and 36.8% was pulmonologist. There were
no differences in terms of age, sex, years of
medical practice, academic positions, trun-
cal specialty and allergy subspecialty;
between physicians who refer and do not
refer their patients for CAM. However,
afliation in a university hospital (11.9%
versus. 53.9%, P = 0.01) was signicantly
associated with a decreased referral for
CAM compared afliation in a state hospi-
tal, private hospital or doctors ofce.
20.8% of the respondents have reported
lack of information whereas 48.8% have
reported little information about CAM.
Acupuncture (40.5%), herbal medicine
(40.1%), massage therapy (27.3%), yoga
(24.8%) and meditation (23.6%) were the
most commonly known CAM methods
among the participants. The most common
allergic diseases for referral to CAM use
were; asthma (42.1%), chronic urticaria
(28.9%), allergic rhinitis (13.3%) and atopic
dermatitis (4.4%).
Conclusion: The current study shows that
academicians seem to be reluctant to refer
their patients for CAM. A small portion of
the physicians believes that they have suf-
cient information about CAM. The physi-
cians should take into consideration the
popularity of CAM use among such
patients.
1702
The unmet need in allergy education
Goh, L1; Patel, N1; Fertleman, C2
1
The Whittington Hospital, Paediatric Department,
London, United Kingdom; 2The Whittington Hospital,
Paediatric Department, London, United Kingdom
Background: Several recent royal college
and government documents have recog-
nised the lack of training for undergradu-
ate and postgraduate health professionals
in allergic disease which in turn impacts on
patient care. We present an overview of a
novel paediatric allergy module for under-
graduate medical students and demonstrate
that this positively impacts on both the
students knowledge of common allergic
disease and the impact on the family.
Methods: Four self-selected, 3rd year Uni-
versity College London undergraduate stu-
dents attended a two-week paediatric
allergy module at the Whittington Hospi-
tal. Members of our multidisciplinary pae-
diatric allergy team led teaching sessions.
A home visit to families of children with
severe allergic disease was also arranged
for the students. The students were
assessed using a case based discussion and
a poster presentation of an allocated litera-
ture research project. The students pro-
vided written feedback for each session. A
pre and post course quiz was set to assess
knowledge and promote learning.
Results: The students completed the
quizzes and assessments. They provided
written feedback on the 9 taught sessions.
The differential results of the pre and post
course quizs and the feedback received
reinforce the existing unmet need in under-
graduate allergy education and the positive
impact this module had towards meeting
this need. The average pre-course result
was 47.5%, and the average post-course
result was 85%.
Conclusion: This small study demonstrates
the incomplete prior knowledge amongst a
self-selecting group of undergraduate medi-
cal students, who all expressed a particular
interest in allergy. Multidisciplinary-based
teaching can effectively address this knowl-
edge gap. In the current allergy epidemic
all doctors require a good basic under-
standing of allergic disease. We propose
that similar allergy modules are incorpo-
rated into undergraduate medical educa-
tion.
1703
Review of allergology departments activ-
ity in the emergency department
Bruno Soares, J; Viegas, L; Mascarenhas, I; Lopes da
Silva, S; Branco Ferreira, M; Lopes, A; Pereira Barbosa,
M
Hospital Santa Maria, Immunoallergology, Lisbon,
Portugal
Background: Patients with allergic manifes-
Department (ED) on a daily basis. The
Allergology Consultation for the Emer-
gency Department was created in 2004,
aiming for the best possible orientation of
those patients.
Aim: To evaluate the referral motives,
treatment and further orientation of the
patients.
Methods: Data collected from the medical
records of all the patients referred by the
641

ED (ALERT), from January 1st 2006 to
December 31st 2010.
Results: A total of 532 consultations were
requested, 374 (70.3%) of female patients,
average age 43.3 18.2 years (median
41.5). The consultations were requested by
Internal Medicine (488; 91.7%); Otorhino-
laryngology (14; 2.6%), Dermatology (24;
4.5%), Stomatology (1; 0.2%) and by
other hospitals (3; 0.6%). The following
were the most frequent motives for consul-
tation: urticaria with or without angioe-
dema (AE) (329; 61.8%), isolated AE (144;
27.1%), and eczema (27; 5.1%). Sixteen
patients had anaphylaxis (3%). Sixteen
patients (3%) were referred for cutaneous
manifestations of non-allergic causes. In
38.2% of the cases the cause was
unknown. In patients with urticaria/AE or
isolated AE, drugs were the probable etio-
logic factor most frequently identied. The
majority of the patients (371; 69.7%) was
treated with antihistamines and systemic
corticosteroids, according to the most fre-
quent manifestations. Thirteen patients
(2.4%) needed epinephrine as well. Most
of the patients (418; 78.6%) were dis-
charged and sent to an Allergology Con-
sultation at the outpatient clinic. Twenty
seven patients (5.1%) were admitted to a
medical ward. Eighty seven patients
(16.4%) were referred to other specialists
(most frequently to their General Practi-
tioner and Dermatologist).
Conclusion: Referral to the Allergology
Department during their ED stay, allows a
faster and better orientation of the allergic
patients. Skin manifestations are the most
frequent referral motives, and a signicant
percentage of the cases are presumably
drug-induced.
1704
Allergy camp: educational intervention
Bellido, V1; Rubio, E2; Lopez, C1; Cimbollek, S3;
Gonzalez-Quevedo, T4
1 Tarragona, Spain
Macarena Hospital Seville, Allergy UNit, Seville, Spain;
2
University of Seville, Psychiatry, Personality,
Evaluation and Psychologic Department, Seville, Spain;
3
Virgen del Rocio, Allergy Unit, Sevilla, Spain; 4Virgen
del Rocio, Allergy Unit, Seville, Spain
Background: Allergy disease is a whole
entity that it has to be understood from a
global point of view: social, medical and
642
psychological. It is a chronic disease with
periods free of symptoms and exacerba-
tions. These exacerbations have different
intensity so this is a difculty added. For
this reason, educational health is very
important in allergy disease.
Methods: Allergy camp takes place during
a week in summer time. The age of chil-
dren is between 7 and 13 years all. This
activity is a great opportunity to improve
in an unnoticed way the knowledge about
self-care of allergy disease.
Hot topics:
(1) Knowledge of own disease.
(2) Trigger factors and avoidance.
(3) Recognise an acute crisis.
(4) Therapeutic resources to control the
crisis.
To carry on with this our team is com-
posed of 23 allergists and one psycholo-
gist. Every day children have a review with
allergists, a workshop with allergists and
the psychologist and sport activities with
rest of children free of illness.
Results: Children are asked before and
after the camp through survey to see the
inuence of these camps on them. In most
of item in survey we can see an improve-
ment higher than a 30% between before
and after the camp.
Conclusion: After medical and psychologist
intervention is observed that children
acquire knowledge of their disease that it
make them feel more condent. At the
same time, they have a better control of
their symptoms and their medication, and
a higher responsibility of their allergy dis-
ease.
1705
Use and handling of self-injectable epi-
nephrine in patients with a history of
anaphylaxis
Andres Lopez, B; Gaig Jane, P; Gazquez Garca, V;
Dalmau Duch, G
Hospital Universitari de Tarragona Joan XXIII,
Background: Self-injectable epinephrine
(SIE) is a treatment that should be pre-
scribed to patients who have suffered an
anaphylactic reaction and are likely to
have another. The aim of this study was to
evaluate the use and management of SIE
in these patients.
Allergy 66, Suppl. 94 (2011): 482642  2011 John Wiley & Sons A/S
Poster Session 74 Allergy care and education
Methods: Patients who were prescribed
SIE and who attend follow-up in our con-
sultations. All of them underwent a 30
question survey about the diagnosis, use
and expiry of the SIE, among others. For
patients under 12-years-old, parents
responded on their behalf.
Results: We included 61 patients (38
women) with a mean age of 37.5 years old
(range 3 to 79). The causes of anaphylaxis
were: food allergy (57.4%), hymenopterous
venom (27.9%) with no identied cause
(9.8%) and exercise-induced anaphylaxis
(1.6%). The time between the reaction and
prescription of the SIE was more than one
year in 41% of patients, and less than one
week in 9.8%. Prescription was made by
the allergist (83.6%), the emergency physi-
cian (8.2%) and primary care physician/
paediatrician (6.6%). Although 47.5% of
patients said they always carried the SIE
with them, only 34.4% had it at the time
of the survey. Of the total patients, 82%
looked at the expiration date. Of these, it
was expired in 24.6% of the cases. Of the
total, 34.4% felt that the price of the SIE
was expensive, 11.5% very expensive and
34.4% were unaware of the price. Only
two patients used the SIE.
Conclusion: We found a signicant delay
in the indication for SIE. The allergist is
the person who prescribes it most fre-
quently. All patients for whom it was pre-
scribed purchased it. It was expired for one
in four patients. They have had to use it
on few occasions.