Vi C ll d Di Viruses, Cells and Disease November 13, 2008 C i ( C ) Human Cytomegalovirus (HCMV) Immediate early proteins, gene expression and signaling QuickTime™ and a decompressor are needed to see this picture. Dr. Hua Zhu ICPH E350D UMDNJ - New Jersey Medical School 973-972-4483 X 2-6488 [email protected]Electron Cryomicroscopy and 3D Reconstruction From Dr. H. Zhou
44
Embed
Human Ci(C)Cytomegalovirus (HCMV)njms.rutgers.edu/gsbs/olc/vcd/prot/2008/Hua Zhu Lecture slides rev.pdfPreparing electro-competent cells ... Model For Studying HCMV Pathogenesis thymus/liver
This document is posted to help you gain knowledge. Please leave a comment to let me know what you think about it! Share it to your friends and learn new things together.
Transcript
Vi C ll d DiViruses, Cells and DiseaseNovember 13, 2008
C i ( C )Human Cytomegalovirus (HCMV)Immediate early proteins, gene expression and signaling
QuickTime™ and a decompressor
are needed to see this picture.Dr. Hua ZhuICPH E350DUMDNJ - New Jersey Medical School973-972-4483 X [email protected]
Electron Cryomicroscopy and 3D ReconstructionFrom Dr. H. Zhou
IntroductionIntroduction1. Medical aspects of HCMV1. Medical aspects of HCMV2. HCMV structure2. HCMV structure3. HCMV replication3. HCMV replication
Special topicsSpecial topics1. IE1 and IE2 functions1. IE1 and IE2 functions2. A HCMV2. A HCMV--induced signal transduction pathwayinduced signal transduction pathway
(di t t k)(di t t k)(discuss two papers next week)(discuss two papers next week)3. Functional profiling of HCMV genome3. Functional profiling of HCMV genome4. Identification of HCMV pathogenic genes4. Identification of HCMV pathogenic genesp g gp g g
Virus• Herpesvirus, dsDNA virus• ~80% of adults are infected• ~80% of adults are infected
Transmission• in utero and at any age
- body fluids• breast feeding, blood transfusion
Latency and Recurrence• Latent in monocytes (Myeloid progenitor cells)• Reactivates during pregnancy and in
immunosuppressed states, resulting in cytomegalic inclusion disease (CID), CMV pneumonia retinitis and hepatitisCMV pneumonia, retinitis and hepatitis
Pathogenicity• In people with cancer, transplanted organs, AIDS or other
immune deficiencies CMV can cause severe diseases of theimmune deficiencies, CMV can cause severe diseases of the lung, liver, colon, eye or brain
Rates of CMV Infection and disease in liver transplant patients according to types of infection
Pretransplant serostatus of
donor/recipientType of infection Infection
(%)Disease
(%)+/+ Reinfection or 66 23+/+ Reinfection or
reactivation66 23
+/- Primary 77 61
-/+ Reactivation 49 10
-/- Primary, not from graft 10 10
HCMV and AIDSC i di t f d th i AIDS (25%)• Common immediate cause of death in AIDS (25%)
• 70-90% show active CMV infection• 30-40% develop CMV retinitis• 95% homosexual men with AIDS have active HCMV infection
CMV retinitis
Congenital HCMV Infection
The most common virus globally to be transmitted in utero,affecting up to 2.5% of all live births (WHO) A leading cause of birth defectsA leading cause of birth defects~10% of infected fetuses exhibit damage of the CNS-mental retardationhearing losshearing lossimpaired vision
Prevention and treatment
• Vaccines: AD169, Towne - not effective
• Antivirals: Ganciclovir - effective
Anti Herpesvirus AgentsAnti-Herpesvirus Agents QuickTime™ and aTIFF (Uncompressed) decompressor
are needed to see this picture.
QuickTime™ and aTIFF (Uncompressed) decompressor
are needed to see this picture.
Acyclovir (Zovirax - Glaxo) - for α herpesviruses Ganciclovir (Cytovene - Roche) - for β herpesviruses
- Guanosine analogs
Mechanism of Action:• Specifically activated by virus-induced kinases,
such as thymidine kinase of HSV and UL97 of CMV• Terminate viral DNA chain elongationTerminate viral DNA chain elongation• Selectively inactivate viral DNA polymerases and inhibit
IE2 is essential for viral replication• IE2 is required for viral early and late gene expression• IE1 and IE2 directly regulate viral gene expression
What Are the Host Response to HCMV?What Are the Host Response to HCMV?
HCMV
gH
gM/N
HSPG
HCMVR
gBg
IEIE
?
Identification of cellular genes altered after HCMV infection
Differential displayDifferential displayGeneChip (Affymetrix)cigs (IFN-stimulated genes, isgs)Activated by both live and killed HCMVNot require viral gene expression, viral DNA,
and new protein synthesisand new protein synthesisNot mediated by IFNs or other cytokinesHCMV directly activates ISGs
cig34
cig6 cig24 cig25
cig41cig1 cig52 cig53
cig43cig42
cig49 control
cig1 = isg54k
ISREGASGAS
What is required for induction of ISGs?What is required for induction of ISGs?
HSPG
HCMV gB
gH
gM/N
HSPG
HCMVR
gH
??
IE
ISG
gM/N
Is fusion required?Is fusion required?
HSPG
HCMV gB
gH
gM/N
PEG
HCMVR
- W W M MHCMV: CFI
isg54K -
cig49 -
CFI: - + +--
7 SK -
HCMV: - + ++-
ISG ?isg54K -
CFI: - - ++-PEG: --
+-+HCMV: + ++
7 SK -
No growth: 45; Severe growth: 12; Moderate growth defect: 23; Growth like WT: 68; Enhanced growth: 4
Dunn et al., 2003, PNAS
Tissue Tropism Tissue Tropism
A: Fibroblast cells; B. Epithelial cells; C. Endothelial cells
CMV (240kb)CMV (240kb)
AD169T
UL US
TowneToledoPH
20 40 60 80 100 120 140 160 180 200 220 230 kb0
TRFIX
The Severe Combined Immunodeficient (SCID) Mice The Severe Combined Immunodeficient (SCID) Mice Model For Studying HCMV PathogenesisModel For Studying HCMV Pathogenesis
thymus/liver tissue
y g gy g g
kidney kidney capsule
3 months
/
Infected implants are harvested at different time
d th t f HCMV A thymus/liver implant is surgically exposed and injected with HCMV
and the amount of HCMV is measured
The clinical but not attenuated strains can replicate in The clinical but not attenuated strains can replicate in ppthe implanted human tissuesthe implanted human tissues