Clinical Immunologic Test

Fubin Wang
Department of Clinical Immunology Test,
Zhongnan Hospital, Wuhan University
Clinical immunologic te
st:

Test of Ig
Test of complement
Test of cellular immunity
Test of tumor markers
Test of autoimmune diseases
Others…
 Test of Immunoglobulin
 Test of serum IgG 、 IgA 、 IgM
 Application of IgG 、 IgA 、 IgM tests
 Test of IgE and IgD
 Test of abnormal Ig
 临床免
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Four subclasses :
IgG1, IgG2, IgG3, IgG4
 Clinical significance
 Ig rise
 1. Polyclonal Ig rise: chronic infection 、 chronic hepatit
is 、 hepatocirrhosis 、 lymphoma and some autoimmu
ne diseases, as SLE 、 RA 。
 2. Monoclonal Ig rise : immune proliferative disease ， as
MM 、 primary macroglobulinemia 。
Ig fall
primary immunodeficiency diseases 、 Acquir
ed immunodeficiency disease and patients with
immunosuppressor 。
Serum IgE ： Clinical significance

1. Type I allergy

2. IgE myeloma 、 parasite infection

3. chronic hepatitis 、 SLE 、 RA

Serum M (monoclonal) protein detect:
 Clinical significance ：
 1. (MM)
 2. Macroglobulinemia 。
 3. Heavy chain disease （ HCDs ）。
 4. Benign Monoclonal Gammopathy (BMG) 。
 5. Malignant Lymphoma 。
 Serum complement detect

 Complement activation (flash)

 Total complement
 50 ％ Hemolytic unit of complement. CH50
Degree of activation of complement
 Rise ： acute inflammation and some maligna
nt tumor.
 Fall ： more significant ， as in some immune
injury diseases: acute glomerulonephritis ， S
LE ， infective endocarditis ， viral hepatitis
。
 Test of C3

 Rise ： acute inflammation 、 early phase of i

nfectious diseases 、 malignant tumor 、 re
jection 。

 Fall ： acute glomerulonephritis 、 SLE 。

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Test of C4:
 Clinical significance are similar as C3 。
 If fall: (MM), IgA nephropathy
Hereditary C4 deficiency
Test of cellular immunity
 Cellular immunity
 Important role in immune system and
protections from pathogens and tumor.
 View flash.
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Detection of T cell CDs
 Cluster differentiation ， CD
 CD3 （ total T cells ）： if fall, autoimmune diseases as SL
E。
 CD4 T cell （ TH ） ： if fall, malignant tumor 、 Her
editary immunodeficiency 、 Viral infection 、 treated
with immunosuppressive agent 。
 CD8 suppressive T cell （ Ts ）： if fall, autoimmune dise
ases or allergy diseases
 CD4/CD8 ratio ： 1 、 if rise ： malignant tumor 、 au
toimmune diseases 、 Viral infection 、 allergy 、 t
ransplant rejection 2 、 if fall ： AIDS
isolation CD4+T cell and CD8+T cell by
FACS
 Test of T cell transformation

 T cells stimulated by PHA or ConA in vitro.

The ratio of transformed T cell reflects the f
unction of T cells 。
 Fall ： low function of T cells as malignant
tumor
 Rise ： hyperthyroidism 、 thyroiditis 、
SLE
 Detection of B cell CDs

 CD19 、 CD20 、 CD22

 If rise: Acute Lymphoblastic Leukemia (A
LL) 、 Lymphoma ；
 If fall: Agammaglobulinemia 、 chemother
apy or immune suppressor 。
 Test of NK cells activation

 If rise: early phase of viral infection ， Do

wn syndrome ， transplantation
 If fall: Malignant Tumor 、 Severe Com
bined Immunodeficiency (SCID) ， AIDS
and therapy of immune suppressor
Test of tumor markers
 Test of tumor markers
 （ Tumor marker): Tumor markers are substances prod
uced by tumor cells or by other cells of the body in respo
nse to cancer or certain benign (noncancerous) condition
s. These substances can be found in the blood, in the uri
ne, in the tumor tissue, or in other tissues. Different t
umor markers are found in different types of cancer,
and levels of the same tumor marker can be altered in mo
re than one type of cancer.
 In addition, tumor marker levels are not altered in
all people with cancer, especially if the cancer is e
arly stage. Some tumor marker levels can also be a
ltered in patients with noncancerous conditions.
 Tumor markers

 Proteins (AFP, CEA, PSA, SCC)

 Glycan (CA19-9, CA125, CA15-3, CA50...)

 Enzyme (NSE, AFU, ACP)

 Incretion (HCG)
 AFP
 Alpha-fetoprotein （ AFP ） AFP is most useful in f
ollowing the response to treatment for liver cancer (hepatoc
ellular carcinoma). Normal levels of AFP are usually less t
han 20 nanograms per milliliter (ng/mL). AFP levels are hig
her than normal in about 2 out of 3 patients with liver cance
r. The level increases with the size of the tumor. In most pa
tients with liver cancer, the level is more than 500 ng/mL
.
 In very small tumors the levels may be less
than 20 ng/mL. AFP is also elevated in acut
e and chronic hepatitis, but is seldom above
100 ng/mL in these diseases.
 AFP is also higher in certain testicular can
cers (embryonal cell and endodermal sinus t
ypes) and is used for follow-up of these can
cers.
 CEA
 Carcinoembryonic antigen （ CEA ） : CEA is the preferr
ed tumor marker for following patients with colorectal canc
er during or after treatment, but it is not useful as a screenin
g or diagnostic test. The normal range of blood levels varies
from lab to lab, but levels higher than 5 nanograms per milli
liter (ng/mL) are generally considered abnormal.
 The higher the CEA level at the time colorectal ca
ncer is detected, the more likely it is that the cance
r is advanced.
 Many doctors use this marker to follow other canc
ers, such as lung cancer and breast cancer. CEA l
evels are also elevated in many other cancers such
as those of the thyroid, pancreas, liver, stomach, o
vary, and bladder.
 PSA
 Prostate-specific antigen （ PSA ） : PSA is a tumor m
arker for prostate cancer. It is the only marker used to scree
n for a common type of cancer (although some medical grou
ps do not recommend its use). It is a protein made by cells o
f the prostate gland in men, which is responsible for making
some of the liquid in semen.
 The level of PSA in the blood can be elevated in p
rostate cancer, but PSA levels can be affected by o
ther factors, too. Men with benign prostatic hype
rplasia (BPH), a non-cancerous growth of the pro
state, often have higher levels. The PSA level also
tends to be higher in older men and those with larg
er prostates, and it can be elevated for a day or two
after ejaculation.
 CA-199
 CA-199: Although the CA 19-9 test was first developed to
detect colorectal cancer, it is more sensitive to pancreatic
cancer. It will not usually detect very early disease, which
is why it is not used as a screening test. But it is now
considered the best tumor marker for following patients
with cancer of the pancreas.
 Normal blood levels of CA 19-9 are below 37 U/
mL (units/milliliter). A high CA 19-9 level in a ne
wly diagnosed patient usually means the disease is
advanced.
 CA 19-9 can also be used to monitor colorectal ca
ncer, but because it is less sensitive than the CEA
test, most medical groups recommend CEA testing
when following this disease.
 CA125
 CA125: CA 125 is the standard tumor marker use
d to follow women during or after treatment for epit
helial ovarian cancer (the most common type of ov
arian cancer). Normal blood levels are usually less t
han 30 to 35 U/mL (units/milliliter). More than 90%
of patients have higher levels of CA 125 when the c
ancer is advanced.
 Levels are also elevated in about half of women who
se disease is still confined to the ovary. Because of t
his, CA 125 is being studied as a screening test. The
problem with using it as a screening test is that it wo
uld still miss many early cancers, and conditions oth
er than ovarian cancer can cause an elevated CA 125
level.
 CA15-3 临床免
疫学检验
 CA15-3: CA 15-3 is used mainly to monitor patients with
breast cancer. Elevated blood levels are found in less than
10% of patients with early disease and in about 70% of
patients with advanced disease. Levels usually drop
following effective treatment, although they may spike in
the first few weeks after treatment is started, a result of
dying cancer cells spilling their contents into the
bloodstream.
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Neuron-specific enolase (NSE):

 NSE: NSE is a marker for neuroendocrine tumors such as

small cell lung cancer, neuroblastoma, and carcinoid tumo
rs. It is not used as a screening test. It is most useful in the f
ollow-up of patients with small cell lung cancer or neurobla
stoma . Elevated levels of NSE may also be found in some
non-neuroendocrine cancers. Abnormal levels are usually hi
gher than 9 micrograms per milliliter (ug/mL).
 HER2
 HER2 (also known as HER2/neu, erbB-2, or EGFR2):
HER2 is a marker that is elevated in some breast cancer
cells. Higher than normal levels are also found in some
other cancers. The HER2 level is usually found by testing a
sample of the cancer tissue itself, not the blood. About 1 in
5 people with breast cancer test positive for HER2. Its main
use is as a predictor of prognosis (outlook for survival).
Those whose cancers are positive for this marker don't
respond as well to chemotherapy, and in the past were
thought to have a less favorable outlook.
 （ human chorionic gonadotrophin ， HC
G）

 Human chorionic gonadotropin (HCG): HCG (als

o known as beta-HCG) blood levels are elevated in patients
with some types of testicular and ovarian cancers (germ c
ell tumors) and in gestational trophoblastic disease, mainly
choriocarcinoma. They are also higher in some men with c
ertain cancers in the middle of their chest that start in the sa
me cells as testicular cancer. Levels of HCG can be used to
help diagnose these conditions and can be followed over tim
e to monitor how well treatment is working. They can also b
e used to look for cancer recurrence once treatment has ende
d.
Test of autoantibodies
 Clinical autoimmune diseases
Autoimmune diseases arise from an overactive
immune response of the body against substances and tissues norma
lly present in the body.

（ Systemic lupus erythematosus ， SLE)

 (Rheumatoid arthritis ， RA )
（ systemic sclerosis, SSc ）
（ Sjogren Syndrome, SjS )
(Mixed connective tissue disease ,MCTD)
Systemic lupus erythe
matosus (SLE)
Systemic Scleroderma
 ANA

 Anti-ENA

 Anti-TCA
 Anti-Nuclear antibody

 Nuclear antigens include DNP (deoxyribonucleic

protein), DNA, RNA and ENA (extracted nuclear
antigens) .

 Different Fluorescences of cell nuclear as Homog

eneous ， (H); (Speckled ， S); Membranous,
(M) ; Nucheolar ， (N)
Fluorescences and clinical significances

homogeneous ， H
（ speckled ， S ）
（ membranous ， M ）
（ nucheolar ，
N）
 ANA positive ： untreated SLE ， 80-90%(+) ， alm
ost 100% in active SLE. Mixed connective tissue disea
se ， (MCTD) 、 Systemic sclerosis (SSc) .

 SLE : homogeneous ， H of Fluorescence

 MCTD: speckled ， S

 SSc: nucheolar ， N
 Anti-RNP antibody

anti-U1-RNP antibody: MCTD ， 30%~

40%(+) in SLE patient.
anti- Mitochondria antibody ( AMA ）
AMA : serum marker of PBC (primary biliar
y cirrhosis)
Anti-Smooth Muscle Antibody (ASMA): marker of autoi
mmune hepatitis ， (AIH) 90 ％ (+);
ASMA(>1 ： 1000) 100 ％

Anti-neutrophil cytoplasmic antibodies, （ ANCA ） :

Systemic Vasculitis
inflammatory bowel disease
connective tissue disease, CTD