EBV Virus Epsteina a Barrov

Gamma herpesvirinae

Pathogenese

 Vtina nemoc spojench s EBV je dsledek latentn infikovanch bunk. To je rozdl oproti HSV-1.
Transmise via sliny, (obvykle v ranch fzch ivota). Infikuje oropharyngaln epitelialn buky a pak vstupuje do B bunk. Cca 1 / 106 B bunk pozitivn na EBV. Infekn mononukleosa (IM): Primrn EBV infekce indukuje proliferaci B bunk , kontrolovanou expans of virus-specifickch cytotoxic T bunk (CTL). V nkterch individuchse vyvine a fatln IM (X-linked lymphoproliferation).

Distribuce Burkitt lymfom

Burkitt, D. Nature 194:232, 1962 Subequatorial Africa

High elevations BL cases

Pathogenese EBV infekce

Vtina nemoc spojen s EBV je zpsobena latentn infekc bunk. Odlinost od HSV-1. en: slinami, v ranm vku (obvykle). Infikuje orofaryngln epitheliln buky a pak vstupuje do B bunk. Cca 1 z 106 B bunk obsahuje EBV. Infekn mononukleosa (IM): Primrn EBV infekction indukuje proliferatci B bunk, co je kontrolovno expans virus-specifickch cytotoxickch T bunk (CTL).

Spojen EBV a BL(Burkittova lymfomu)

BL v oblastech Afriky s vysokm vskytem moskyt vedlo k mylence, e tumor by mohl bt zpsoben moskyty penenm virem To vedlo Epsteina, Barrovou a Achonga vyetovat BL buky na ptomnost vir. Eletronovm mikroskopem objevili v r. 1964 EBV kter se tak stal prvnm kandidtem na virus psobc lidk ndorov onemocnn. Nsledn studia ukzaly, e EBV je ptomen ve 100% BL .

Burkittv lymfom

Na rozdl od immunoblastickch lymfom, BL je pomaleji vznikajc monoklonln tumor multifaktorov etiologie.

EBV infekce je nezbytn avak nedostaujc pro vvoj BL. Kofaktory - imunosuprese a genetick udlosti (chromosomln translokace).

Burkittv lymfom
EBV+: 90% ppad v rozvojovch zemch AIDS pacienti tumory lymfatickch uzlin

EBV + . Vechny tumory maj c-myc translokace

Deregulation exprese c-myc V nkterch pouze EBV EBNA-1 exprimovna

Terapie : Chemoterapie

Multifaktorov etiologie Burkittova lymfomu

EBV
B

Malaria

Genetic Event
BL

polyclonal B-cell activation, proliferation, immortalization Resting B cell

polyclonal B-cell activation T-cell immunosuppression

Chromosomal translocation & c-myc activation Transformed B cell

Activated, proliferating B cells

Tumory indukovan EBV

Tumor
Burkitts lymphoma

Subtype
Endemic Sporadic AIDS-related

Latent Period
3-8 years post EBV 3-8 years post EBV 3-8 years post EBV > 30 years post EBV > 30 years post EBV > 30 years post EBV > 10 years post EBV < 3 months post EBV

EBV Positivity
100% 15-85% 30-40% 100% 30-100% 100% 20-80% 100%

Nasopharyngeal carc inoma T cell lymphoma Hodgkins disease Posttransplant lymphoproliferative disease (PTLD)-like lymphomas

Nonkeratinizing Keratinizing Nasal Various Fatal IM

Tr-associated AIDS-related

< 1 year post Tr > 8 years post HIV

>90% >80%

BL: Chromosomln translokace

Ig-Hvy Chain: Rearranged; Actively expressed CS14 CS8 E I II III

c-myc Gene: Exons Translocations: lose exon I (neg. ctrl?) 8/14

E
E I

II II

III III

control by Ig-Hvy Enh Other translocations: CS22: Ig-Lite Chain l enhancer; CS2: Ig-Lite

k enhancer

Nemoce spojen s EBV

EBV v B bukch Infectious mononucleosis X-Linked Lymphoproliferative Disease Chronic active EBV Hodgkin Disease Burkitt Lymphoma Lymphoproliferative disease EBV v jinch bukch Nasopharyngeal carcinoma Gastric carcinoma Nasal T/NK cell lymphomas Peripheral T cell lymphomas Oral hairy( nebezpen) leukoplakia Smooth muscle tumors in transplant patients

Hodgkinova choroba

EBV+: 60-70% ppad v rozvinutch zemch

35-50% ppad v USA Terapie: Chemoterapie, ozaovn Anti-EBV CTLs efektivn v nkterch ppadech

LMP-1 exprese

EBV pozitivn tumory hladkch sval

U transplant recipient, AIDS patient
Pathology: sarkomy a myomy rznch orgn (zejmna transplantt ) a lymfatickch uzlin

EBV Lymphoproliferative Disease

Pi imunodeficienci transplantacch (AIDS ) po Symptomy: Infekn mononukleosa Masivn lze v orgnech (mn asto v lymfatickch uzlinch ) Rizikov faktory: Primrn infekce vzrstajc imunosuprese

CMV

Cohen NEJM 2000

Rizika EBV PTLD (proliferative T lymfocyte disease)

Primrn infekce vy dvky viru, neptomnost T bunk specifickch k EBV Infekce cytomegalovirem Polymorfismus odpovdajc nzk produkci interferonu IFN-, a tumor nekrosis faktoru TNF-; vysok hladiny interleukinu IL-10 munosuprese T bunk

Biologick vlastnosti EBV

Infekce primrnch B- bunk - nejprve ustaven latence v sti latentn infikovanch bunk pot indukce lytickho cyklu aktivac transkripnch faktor - produkt -gen. Lytick prbh pak dle podobn jako u herpes simlex viru Latentn infekce primrnch B lymfocyt Latentn infekce primarnch B bunk. Virus infikuje nedlc se B buky s velmi vysokou innost. Indukuje a udruje bunnou proliferaci - imortalisuje je .

Genov exprese v latenci. Zhruba 10% EBV genomu je exprimovno bhem latence. Rozdl oproti HSV-1. V latentn infekci lei schopnost stimulovat a imortalisovat B buky.

Receptory Gammaherpesviruses
EBV: gp350/220 ve komplement receptor 2 (CR2, CD21); KSHV: K8.1A je pozin homolog EBV gp350/220 a ve se k heparan sulftu. gB se ve tak k HS a k integrinu, 31.

Spear & Longnecker. J. Virol. 77:10179, 2003

Poten kroky EBV infekce B bunk

Adsorpce na povrch bunk & vstup Virus se ve k CD21. Cross-linking CD21 spojeno s vyslnm signlu do B bunk progrese bunnho cyklu & upregulace molekul adhese shlukovn bunk Autocrinn sekrece rstovch faktor Shlukovn bunk zvyuje lokln hustotu bunk reakce na sekretovan rstov faktory, kter jsou upregulated v odpovdi na virovou genovou expresi. In vitro EBV transformuje B buky a vytv tak lymfoblastoidn bunn linie (aktivovan, proliferujc & imortalisovan)

Lytick Cyklus EBV

Oropharyngeal epithelialn buky: EBV infekce tchto bunk je lytick (produktivn).
B buky: Mal proporce latentn infikovanch B bunk spontnn indukuje lytick cyklus kontinuln tvorba infeknho viru. Reaktivace: Klov virov regultor pechodu z latence do lytickho cyklu je produkt genu BZLF-1 (protein ZEBRA nebo Zta), siln transaktivtor bezprostedn asnch gen.

Replikan cyklus EBV : Role Zta (ZEBRA)

LYTIC CYCLE

EBV Infection
Cellular activator or repressor

LATENCY

Immediate early genes expressed

Immediate early genes not expressed

Early and late genes expressed

EBNA-1 expressed (LMPs, EBNAs) S timuli (AP-1) BZLF-1 (ZEBRA) expression

Productive Infection

EBV Infekce
Adapted from Kieff and Rickinson, Fields Virology, 3rd Ed., Chapter 75

Primary Infection
Primary infection of epithelium

Persistence
Secondary infection of epithelium

Reactivation
Lytic

Lat EBV

Lat EBV

Lat EBV

Lat EBV

Resting B cell

Lat EBV

Kill
CTL

Kill

Resting T cell

CTL

Memory T cell

Molekulrn aspekty latence

Exprese EBV gen v latenci

EBV vnuje pomrn velkou kapacitu gen na ustaven a udrovn latentn infekce v B bukch. Vechny geny porchzej sestihem Geny exprimovan v latenci : nejmn 12: EBNAs: EB nuclear antigens; EBNA-1, 2, 3A, 3B, 3C & LP LMPs: latent membrane proteins. LMP 1, 2A, 2B EBERs: EB-virus early RNAs. Mal RNA nekodujc protein (x2). BARTs: Vysoce sestien BamA transkripty- koduj BARF0- mRNA

LMP 2

LMP 1

Typy genov exprese EBV v latenci

Klidov B buky: EBV me pet bez exprese jakhokoliv proteinu. Typ I latence: BL EBERs, BARTs and EBNA1 (pouze). Type II latency: NPC tumory LMP (plus EBERs, BARTs, EBNA1) Type III latency: produkovan Vechny latentn proteiny jsou

Exprese latentnch gen EBV

Latence Type 1 2 3 EBER EBNA-1 EBNA-2 EBNA-3 LMP-1 LMP-2 + + + + + + + + + + + + Nemoc BL NPC, HD IM, LPD

jin

+/-

+/-

Nosi

oriP
Latentn origin: Origin DNA synthesy pro episomln EBVv latentn infikovanch B bukch.
Episome segregation: oriP schopnost pomoci segregaci replikujcch se episom do dceinnch bunk bhem bunnho dlen. EBNA 1 operuje v oriP.

EBNA1 poadovna pro replikaci plasmid z ori P

Frappier and O'Donnell, PNAS 88:10875, 1991; Su et al. PNAS 88:10870, 1991

OriP: Action v oriP Akce EBNA1

enhancer/ FR (family of repeats) ~1000bp 20 FR contains multiple high affinity EBNA1 binding sites EBNA-1 EBNA-1 binding sites

DS (dyad symmetry element)

The DS site corresponds to the initiation site for DNA replication. However, EBNA1 binding here is weak and easily dissociated

EBNA1 binds to FR. A high local concentration of EBNA1 results, allowing EBNA1 to bind to DS.

EBNA-1 binds both DS and FR, causing DNA looping

Mitotic chromosome spreads BL41 (EBNA1 neg) Raji EBNA1

EBP-2

DAPI

Overlay B/D EBP-2 a EBNA1 kolokalizuj na hostitelskch mitotickch chromosomech

Lidsk Herpesviry
Alpha herpesviruses: herpes simplex virus-1 herpes simplex virus-2 varicella zoster virus Beta herpesviruses: cytomegalovirus human herpesvirus-6 human herpesvirus-7 Gamma herpesviruses:

Epstein-Barr virus human herpesvirus-8

HHV 8
Kaposis Sarcoma associated Herpesvirus

Herpesvirinae
K.Sarkom u HIV pacient pevn u tch , kte zskali HIV sexuln cestou ( ne u tch kte skali HIV z krevnch prepart nebo u dt) HHV 8 objeven 1994 Lymphocryptovirus (EBV) Rhadinovirus HHV8

HHV 8 isolty

Prevalence of KS in various HIV+ populations

Hemophiliac Heterosexual IV drug user Transfusion recipient Homo- or bisexual
0 5 10 15 20 25 Male Female

% of AIDS Patients with KS

Beral et al. Lancet 335:123, 1990.

HHV-8 Related Malignancies

Multicentric Castlemans Disease

Kaposi's Sarcoma

Primary Effusion Lymphoma B cell (clonal)

B cell (polyclonal)

Endothelial
Moore & Chang, 2000

Multicentric Castlemans Disease

vIL-6 immunolocalization

B cell lymphoproliferative disorder ~50% MCD patients KSHV+ ~80% fatality B cell proliferation due to vIL-6
Moore & Chang, 2000

Malignance spojen s HHV-8

Cesarman. NEJM 349:1107, 2003

Pathogenese Kap. sarkomu

nutnost kofaktoru
Ne typicky neoplastick buky: nemoc hyperproliferativn ne neoplastick per se spe

Spindle buky endothelilnho pvodu : produkuj angiogenn & rst podporujc faktory, pouze 1-5% tchto bunk produkuje proteiny spojen s lytickm cyklem HHV8 . Vtina je latentn infikovanch. Latentn geny zahrnuj kaposin a ORF 71-73 (vFLIP, vCYC, LANA) Vrazn zven vskyt u HIV+ osob: kofaktory nebo HIV proteiny (jako Tat) by mohly pispvat KS pathogenesi

Kaposiho sarcom

Ke: erven skvrny

erven skvrny na gastritic. mukose

HHV-8 - SPOJEN S NEOPLASI

Kaposiho sarkom (KS): klasick endemick spojen s HIV post-transplantan Primarn lymfomy (PEL) Multicentric Castlemans disease (MCD): plasma cell variant hyaline-vascular variant

OTHER HHV-8 ASSOCIATION WITH NEOPLASTIC DISORDERS

Angio-immunoblastic lymphadenopathy Squamous cell skin carcinoma of transplant recipient Angiosarcoma Cutaneous T-cell lymphoma Multiple myeloma Benign monoclonal gammopathy

% S PROTILTKAMI K HHV-8 LATENTNCH A LYTICKCH ANTIGENU VE VZORCCH 246 DT V ALEXANDRII, EGYPT

% s protiltkami k HHV-latentnm proteinm

% s prtotiltkami k HHV-8 lytickm proteinm

60

11

50

40

30

20

10

0
13 46 79 1012 >12

-1

HHV-8 lytick

HHV-8 latentn Andreoni M et al

% S PROTILTKAMI K HHV-8, HHV-6, EBV a CMV VE VZORCCH 246 DT V ALEXANDRII, EGYPT

100 80

60
40 20 0

13

46

79

1012

>12
EBV CMV
Andreoni M. et al

HHV-8 latent

HHV-8 lytic HHV-6

Pedpokldan zpsoby transmise

Sexuln Horizontln Z matky na dt Organov i tkov transplantty Krevn transfuse

Pedpokldan zpsoby transmise

Rizikov skupiny % HHV-8 seropositivn Rizik. skup. Kontroly
2260

Homosexuln mui

0-20 9
0-3

Prostitutky
STD clinics (heterosexual. HIV-pozitivn)

36 5-9

Pedpokldan zpsoby transmise

Horizontln
Detekce protiltek a HHV-8 sevenc v PBMC dt Pattern rstu protiltek u dt v endemickch oblastech, kter ppomn patern rstu jinch herpetickch vir

Detekce HHV-8 ve slinnch lzch a oropharynx v nkterch studich

V Sardinii seroprevalence protiltek k, HHV-8 je 39% u roddinch len s KS, ve srovnn k 11% v kontroln

populaci

Pedpokldan zpsoby transmise

Transplantace: 1 ppad dokumentovanho penosu transplanttu ( orgnu) na recipienta Krevn transfuse: detekce HHV-8 sekvenc v PBMC krevnch drc

Pedpokldan zpsoby transmise

Z matky na dt Vy prevalence protiltek k HHV-8 u dt narozench seropositivnm matkm

DETEKCE HHV-8 POMOC PCR V RUZNCH POPULACCH

Populace
UK a Francie zdrav kontroly USA - zdrav kontroly Zdrav Italsk individua HIV infikovan, zdrav jedinci v Cambii Krevn drci centrl. Afriky Dti v Zambii 10% 23% 8%

PBMC Lymphoid tkn

0 3% 10%

Sperma
0

1323%

Vstup HHV 8 do bunk

Heparan sulft ?? Koncentrace virion na membrn ?? Integriny ?? gB m RGD sekvenci Cystin/glutamt transportr ?? (Science 2006) V obalu - 7 povrchovch glykoprotein pouze 3 ( gB, gH, gL) poadovny pro receptorem zprostedkovanou fzi

Signln transdukce spojen s HHV vstupem do bunk FAK PI3 kinasa MEK / ERK

Srovn genom EBV a HHV8

Damania, B. Nature Rev Microbiol 2:656, 2004

11 virovch mRNA lyt. fze inkorporovno do virion bhem morfogenese ty jsou pipraveny pro okamitou translaci po vstupu viru do bunk

1 -3% infik. bunk projde lyt. cyklem, ostatn ustav latenci v bukch (B lymf. I dalch endothelilnch bukch, ale i ve fibrobl . a epithelilnch bukch)

Latence
Latentn transkripty oblast mezi orf K12 a orf 74

Latentn genov produkty a jejich funkce

ORF 73 - Latency associated nuclear antigen LANA,
velk multifunkn prot. , serologick marker Obdoba EBNA 1, ustaven episom. replikace v lat. infik bukch deregulace bunnho rstu, ve p53 i Rb pozitivn i negativn regultyor transkripce ( i hostitelsk )

v cyclin vir. Homol c-cyklinu D

v FLIP produkt ORF1 homolog bunnhoFLICE inhibition proteinu
Fas associ.death domain - like interleukin 1 beta converting enzyme nyn caspase 8

ve TRAF2 (tumor necrosis factor receptor assoc. Factor 2 ) Jun aktivace

Interferuje s casp8 funkc; jeho exprese apreguluje antiapoptotick transkrip, faktor NFkappaB.

 Kaposiny - nejmn 3 proteiny m RNA K12 ORF Kaposin A mal hydrofobn protein 60 aa (vzan k membrnm) - ve G exchange faktor
regultor integrinem zprostedkovan adhese Kaposin B ( v oblasti DR1 a DR 2), rozpustn protein jadern dal protein.produkty a peptidy - funkce ne zcela jasn aktivace MAP kins

LANA 2 exprese pouze v B bukch inhibuje interferonovou indukci redukovan aktivace protein kinasy R

Molekulrn aspekty rstov deregulace Gamma herpesviry

Transformujc proteiny kodovan prvnmi ORF EBV & KSHV

LMP1 & K1 aktivuje signal. transdukci B- bunk via bunn faktory

Damania, B. Nature Rev Microbiol 2:656, 2004

Bunn efekty protein modulujcch signalisaci LMP2A and KSHV15

Tyto proteiny blokuj signln drhu z BCR (B cell receptor) a pomhaj udrovat latenci

Damania, B. Nature Rev Microbiol 2:656, 2004

Gammaherpesvirov Cytokines & Chemokines

vIL10 & vIL6 indukuj proliferaci B bunk; tak zprostedkovvaj imun. nik
Damania, B. Nature Rev Microbiol 2:656, 2004

HHV-8
Virus zskal adu bunnch gen HHV 8 obsahuje geny kter koduj proteiny s funkc podobnou jako maj hostitelsk proteiny v transformaci anti-apoptose angiogenesi negativn regulaci interferonu

HHV-8 HOMOLOGY BUNNCH PROTEINU KONTROLY BUNN PROLIFERACE

Virov homology orfK1 orfK2 (vIL6) orfs K4, K4.1, K6 orf 16 (vBcl-2) orfK9 (vIRF) orf K13/orf, 71(vFlip) orf 72 (vCyclin) Bunn homology Ig light chain IL6 MIP I, I, II Bcl-2 IRF-2 DEDs Cyclin D2 Funkce Transforming in rat fibroblasts Cell proliferation Induce angiogenesis, inhibit monocytes chemotaxis Anti-apoptosis Negative regulation of interferon-mediated signalling Antiapoptosis Cell cycle control (G1 to S), transforming properties

HHV-8 vIL-6 je indukovn IFN a m opan inky ne IFN

Virov deregulace bunnho cyklu

vCYC

Virov homolog cyclinu D.

Podobn jako bunn cyklin D, se ve do komplexu s cyclin-dependentn kinasou, CDK6. Ale na rozdl od bunnho homologu, vCYC brn normln akumulaci bunk v klidov G1 fzi bunnho cyklu i kdy jsou vystaveny podmnkm , kter by mly vst k zastaven rstu.

LANA

Latency-Associated Nuclear Antigen M klovou roli v deregulaci bunnho rstu.

IRD - Internal Repeat Domain, bohat na vyznaen aminokyseliny

Dourmishev et al. Micro Mol Biol Rev. 67:175, 2003

LANA blocks serum-deprivation induced cell death

Apoptosis (%)
50 40

LANA blocks apoptosis

30
20

10
0

Vector

LANA

Friborg et al. Nature 402:889, 1999.

Coordinovan interference s Rb a p53

Cyclin/CKI pathways vCYC

X RB X
Control of Cell Cycle Progression

p53

LANA vIL-6 v Bcl-2 vFLIP

vIRF

c-myc

X
Induction of Apoptosis

Antman & Chang. N. Engl. J. Med. 342:1027, 2000.

KSHV psob globln reprogramovn bunn transkripce

Transkripn reprogramovn KSHV

KS Expression Signature

erven geny upregulovny

Wang et al. Nature Genetics 36, 687 - 693 (2004)

KSHV psob reprogramovn transkripce endothelilnch bunk

Infection by KSHV reprograms the transcriptome of lymphoid endothelial cells (LEC) and blood endothelial cells (BEC), bringing the two closer together. It is the first virus known to globally reprogram the EC transcriptome.
Wang et al. Nature Genetics 36, 687 - 693 (2004)

Lidsk Herpesviry
Alpha herpesviruses: herpes simplex virus-1 herpes simplex virus-2 varicella zoster virus Beta herpesviruses: cytomegalovirus human herpesvirus-6 human herpesvirus-7 Gamma herpesviruses:

Epstein-Barr virus human herpesvirus-8

Beta herpesvirinae: cytomegalovirus (HCMV) human herpesvirus-6 human herpesvirus-7 my cytomegalovirus

rody: Cytomegalovirus, Roseolovirus Muromegalovirus

Dlouh reprodukn cyklus, infekce v tkov kultue velmi pomal Infikovan buky asto zvten cytomegalia. Virus me bt udrovn v latentn form v sekrencg lzch, lymforetikulrnch bukch ledvinch , pp. jinch tknch. Lidsk fibroblasty uvny pro isolace viru Genomy 145 - 162 kbp 196 - 241 pro roseoloviry (84 -85 genovch produkt) pro cytomegaloviry (166 genovch produkt

Geneticky rznorod podele, mal mra homologie

CMV, HCMV
Typick struktura jako u ostatnch herpesvir, vt v prmru 200-300 nm Tegument mnoho protein i bunn a virov RNA Obalen membrnou odvozenou ER Golgi intermediate compartmennt (ERGIC) Obsahujc nejmn 20 virem kodovanch glykoprotein

GENOM HCMV

4 isomery

Druhov specifick Normln hostitel mononukleosa cca 8% ze vech IM ppad asto bezpznakov infekce zdka pneumonie hepetitis, CNS onemocnn tk a fatln onemocnn nepijmut transplantt

Imunosuprrimovan

en:

kontakt osoba-osoba

Kongenitln abnormality

orofaryngln sekrece mo, cervik. a vaginln sekrece mlko slzy, krev - infekce v dloze - pokozen mozku kongenitln

Mentln retardace, ztrty sluchu, kalcifikace perivengrikulrn mikrocefalie HCMV a imunitn systm Autoimunitn choroby

Hydrocephaly
Splenomegaly + hepatitis

HHV 6 - primrn infekce exzanthem subitum HHV6A HHV6B - Lymfocyty HHV 7 CD4+ T buky Chronick navy Neoplasie ( lumffomy Multiple sclerosis (HHV6) Hypersensitivita k lkm CD4, NK DC, astrocyty PBMC

Cytomegalovirus
RNDr K.Roubalov CSc. NRL pro herpetick viry

Lidsk cytomegalovirus
-herpesviridae, nejvt HV, cca 200 gen Pbuzn viry: my, krys, opi, morec Kosmopolitn rozen, vysok promoenost populace (50-90%)

NRL pro herpetick viry, SZ, Praha

irok spektrum hostitelskch bunk

Divok kmeny: makrofgy, endotelie,lymfocyty, granulocyty,epiteliln buky, fibroblasty Latentn infekce: kmenov buky,prekursory myeloidn ady Laboratorn kmeny ztrc schopnost infikovat endotelie a leukocyty Mno se na lidskch embryonlnch fibroblastech nebo epitelilnch bukch

ivotn cyklus CMV

NRL pro herpetick viry, SZ, Praha

Interakce s imunitnm systmem

Inhibice exprese MHCI,II na povrchu infikovan buky gpUS2,3,6,11 Virov protein homologn s MHCI gpUL18 Homology receptor pro CC- chemokinyUL33,78,27,28 Homology receptoru pro Fc IgG- UL118,119 Homology cytokn a chemokn vIL-10,vCXCI (vIL-8) Inhibitory apoptzy: IE1,IE2, v-ICA, v-MIA
NRL pro herpetick viry, SZ, Praha

Diagnosticky vznamn antigeny

Pp72-IE1(MIE): nestrukturln, regulan, bezprostedn asn jadern antigen, zahajuje replikan cyklus viru, asn prkaz infekce bunk Pp65- strukturln tegumentov protein, nadprodukovn pi replikaci virustice, hlavn antigen pro CTL, akumuluje se v granulocytech
NRL pro herpetick viry, SZ, Praha

Dal dleit proteiny

gpB obalov glykoprotein, interaguje s heparan-sulftem, genotypizace UL54 DNA polymerza + UL97 protein kinza dleit pro terapii Pp150 hlavn kapsidov protein

NRL pro herpetick viry, SZ, Praha

Patogenita CMV
Imunokompetentn pacienti: Mal dti: hepatitis, gastroenteritis, pneumonie, konjunktivitis Star jedinci: infekn mononukleosa (10%) Imunodeficientn pacienti:
Novorozenci:
kongenitln infekce

Transplantovan pacienti:
Intersticiln pneumonitis gastroenteritis hepatitis-hepatosplenomegalie sepse, pancytopenie GVHD, rejekce tpu atheromatoza u t.srdce

NRL pro herpetick viry, SZ, Praha

HIV+: retinitis gastroenteritis, esofagitis encefalitis-encefalopatie

Kongenitln infekce CMV

Vskyt: 0,5-2% iv narozench dt Primrn infekce matky: Maximln riziko-v prvnm trimestru thotenstv Rekurentn infekce a u 25% seropozit. thotnch, asymptomatick vyluovn viru do cervikl. sekretu Minimln riziko infekce plodu bezpznakov
NRL pro herpetick viry, SZ, Praha

Relativn riziko kongenitln infekce CMV

IgM protiltky proti CMV u matky 13% Serologicky prokzan primoinfekce 2040% Prkaz CMV DNA v plodov vod 100% Symptomatick infekce u cca 50% kongenitln infikovanch dt

Penos infekce CMV mateskm mlkem

Vyluovn CMV do kolostra: Lokln reaktivace infekce pi laktaci U 90% seropozitivnch en 1-30 tdn po porodu (max. 2-3. Tden)
Riziko infekce pro dt: Nakaz se cca 25% dt Riziko zvis na mnostv vyluovanho viru a dob vyluovn U normlnch dt infekce bezpznakov Nedonoenci u 12% septick stav, pneumonie, hepatosplenomegalie Prevence: tepeln inaktivace, zmraen na 20C

NRL pro herpetick viry, SZ, Praha

Diagnostick znaky infekce CMV

Specifick protiltky IgG,IgM,IgA Pp65 antigenemie Viremie Virurie Detekce virov DNA (mRNA)

NRL pro herpetick viry, SZ, Praha

Doporuen postupy v diagnostice CMV Imunokompetentn pacienti:

Prenatln diagnostika kongenitlnch infekc: Prkaz primoinfekce u matky: serologie,prkaz virov DNA v perifern krvi Prkaz infekce plodu: Detekce CMV-DNA v amniotick tekutin Infekce u novorozenc: Serologie,detekce CMV v moi a v perifernch leukocytech, prkaz virov DNA v likvoru Infekce u malch dt: Serologie, prkaz virov DNA v plasm, prkaz CMV ve vtru (nosohltan, spojivka) Infekn mononukleosa: Serologie, prkaz virov DNA v plasm Neurologit pacienti: prkaz virov DNA v likvoru
NRL pro herpetick viry, SZ, Praha

Doporuen postupy v diagnostice CMV

Imunodeficientn pacienti:
Preventivn sledovn aktivn infekce: Prkaz antigenemie Kvantitativn prkaz CMV-DNA v perifern krvi Diagnostika cytomegalovirovho onemocnn: Prkaz antigenemie Kvantitativn prkaz CMV-DNA v perifern krvi Detekce CMV-DNA v biopsii Prkaz CMV-DNA v BAL, likvoru, on tekutin
NRL pro herpetick viry, SZ, Praha

Diagnostick metody
Srologie: IgG,IgM,IgA Pm prkaz: Klasick izolace na TK Zrychlen izolace (Shell vials) prkaz antigenemie pp65 v krvi Prkaz virov DNA (kvantitativn) Prkaz virov mRNA
NRL pro herpetick viry, SZ, Praha

Antigen CMV (pp65) v leukocytech

NRL pro herpetick viry, SZ, Praha

Detekce CMV v kultue lidskch fibroblast (prkaz asnho antigenu)

NRL pro herpetick viry, SZ, Praha

Terapie
Preventivn Preemptivn Symptomatick Antivirotika: gancyklovir, foscarnet, cidofovir (hyperimunn IgG) Vakcny: ve vvoji

NRL pro herpetick viry, SZ, Praha

Konec 2. sti