Download - H. S TOIBER V IENNA, 19.07.2010 MULTIFACETED INTERACTIONS OF RETROVIRUSES WITH THE COMPLEMENT SYSTEM
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Virologie
H. STOIBERVIENNA, 19.07.2010
MULTIFACETED INTERACTIONS OF RETROVIRUSES WITH THE COMPLEMENT
SYSTEM
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SCHEMATIC OVERVIEW OF IMMUNE RESPONSES UPON VIRAL INFECTIONSVienna, 19.07.2010
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Complementactivaton
MACLysis
Infected cellPathogens
Inflammation
Chemotaxis
C3aC5a
IncreasedPermeability
C3aC5a
APCsGranulocytes
BasophilsMast cells
Antigen -Presentation
T cell
DCs
Opsonisation
Immune complexes
CR3
CR1
M
Phagocytosis
YCR2
FDCs
Trapping
Cell activation
BasophilsMast cells
Granulocytes
DCs
YY Y
B cells
C3aC5a
iC3bC3d
CONSEQUENCES OF COMPLEMENT ACTIVATIONVienna, 19.07.2010
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(Schacker et al. 2000)
EX VIVO DETECTION OF HIV IN LT OF INFECTED PATIENTS Vienna, 19.07.2010
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COMPLEMENT-DEPENDENT TRAPPING OF HIV IN THE LT
Kacani et al. JV
Vienna, 19.07.2010
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Species Virus Source Trapping AIDS-like Symptoms
African Green Monkey SIVagm no no
Sooty mangabey SIVsm no no
Rhesus Monkey SIVmac yes yes
Chimpanzees SIVchp/HIV-1 no no
Human HIV-1 yes yes
RELATIONSHIP BETWEEN TRAPPING AND AIDS Vienna, 19.07.2010
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Complementactivation
MACLysis
Infected cellsPathogens
Inflammation
Chemotaxis
C3aC5a
C3aC5a
APCsGranulocyte
BasophilsMast cells
Antigen -Presentation
T cell
DCs
Opsonisation
Immune complexes
CR3
CR1
M
Phagocytosis
YCR2
FDCs
Trapping
Cell activation
BasophilsMast cells
Granulozytes
DCs
YY Y
B cells
C3aC5a
iC3bC3d
CONSEQUENCES OF COMPLEMENT ACTIVATIONVienna, 19.07.2010
IncreasedPermeability
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IN VITRO PRIME-BOOST EXPERIMENTS: PROTOCOL
Loading of DCs withSEB, HIV, HIV-C autologous, unstimulated CD8+ T cells (Prime)
Boost of primed CD8+ T cells with Ag-loaded DCs (3x)(all cells from the same donor)
Data-Acquisition of Prime-Boost Experiments
Proliferation-assays(CFSE staining)
IFN-g assays (ELISA, Secretions-assay)
Test on Specificity and Functionality ofin vitro generated CTLs
(Tetramer, degranulation, antiviral activity)
Vienna, 19.07.2010
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Bánki et a., PLoS Path., 2010 Apr 29;6(4):e1000891.
PROLIFERATION OF HIV-C-DC-PRIMED CD8+ T CELLS (7 DONORS 5 VIRAL STRAINS) Vienna, 19.07.2010
Control (mDCs) HIV-hiC (X4) HIV-C (X4) HIV-C (X4)-P2 HIV-C (R5X4) HIV-C (R5)
0.1%1.5% 7.6% 1.1% 7.2%14.2%
CD8+ T cells
Tetramer-staining
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ANTIVIRAL RESPONSE OF HIV-C-DC-PRIMED CD8+ T CELLS(5 DONORS, 4 VIRAL STRAINS)
Vienna, 19.07.2010
Bánki et a., PLoS Path., 2010 Apr 29;6(4):e1000891.
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Vienna, 19.07.2010ERYTHROLEUKEMIA INDUCED BY FRIEND VIRUS (FV) IN MICE
1st StageExpansion of erythroblastsin spleen due to gp55
2nd StageMalignant transformation of erythroblasts
LTR gag pol env LTR LTR
env: Recombinationdeletion, insertion
F-SFFV F-MuLV
gp55
LTR gag pol env
(replication competent helper-virus)
1. Helper virus, F-MuLV (murine leukemia virus), is responsible for growth of the virus complex.
2. F-MuLV can only produce lymphomas if injected into neonatal mice.3. F-SFFV (spleen focus-forming virus) is defective/interfering virus that expresses
gp55 env protein that binds to EpoR to produce erythroblastosis; has no oncogene;
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%in
fect
ed c
ells
40
30
20
10
0
0.0405
B6 contro
l
B6C3-/- co
ntrol
B6 7dpi
B6C3-/- 7dpi
10
8
6
4
2
0
CD8+
%FV
gag
tetr
amer
+ 0.0119
B6 contro
l
B6C3-/- co
ntrol
B6 7dpi
B6C3-/- 7dpi
8
6
4
2
0
CD8+
%FV
gag
tetr
amer
+CD
43+
0.0088
B6 contro
l
B6C3-/- co
ntrol
B6 7dpi
B6C3-/- 7dpi
ENHANCED FV-INFECTION OF C3-DEFICIENT MICE CORRELATES WITH LOWER FREQUENCIES OF FV-SPECIFIC CTLS.
Vienna, 19.07.2010
Bánki et a., PLoS Path., 2010 Apr 29;6(4):e1000891.
B6 contro
l
B6C3-/- co
ntrol
B6 4dpi
B6C3-/- 4dpi
5
4
3
2
1
0
CD11
c+ %
infe
cted
cel
ls
0.0025
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Complementactivation
MACLyse
Infected cellsPathogens
Inflammation
Chemotaxis
C3aC5a
IncreasedPermeability
C3aC5a
APCsGranulocyte
BasophilsMast cells
Antigen -Presentation
T cell
DCs
Opsonisation
Immune complexes
CR3
CR1
M
Phagocytosis
YCR2
FDCs
Trapping
Cell activation
BasophilsMast cells
Granulocyte
DCs
YY Y
B Zellen
C3aC5a
iC3bC3d
CONSEQUENCES OF COMPLEMENT ACTIVATIONVienna, 19.07.2010
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COMPLEMENT ACTIVATION, MAC AND OPSONISATION
Host cells are protected from CML by Regulators of the Complement system
factor I
+ CR1+ factor H+ MCP
P
iC3b
factor I
C3d
P
CD59
P
P
Complement Activation
MAC
C3
P
C3b
LYSE
CD55fH
Why are intact and infectious viruses in the serum?
Vienna, 19.07.2010
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EfficientVirolyse
InefficientVirolyse
+ Serum + Serum + Antibody
Cell infected with Retroviruses
“budding”gp120/41 complex
MHC-I
MHC-II
DAF
CD59
fH
Stoiber et al. JEMStoiber et al, Nat. Med.
Vienna, 19.07.2010 BUDDING OF HIV
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1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20Factor H
C3b bindingcatalytic region C3b/c binding
heparin binding heparin binding
C3b/d binding
2 7 13 18 19 20Expression
SCR13
Cross-link to mAb
SCR7SCR2 SCR20
SCR18 SCR19
FACTOR H IN DETAIL
Binding site is distinct from the enzymatic active site
Pathogen-specificAntibody
Complement-specificPart
Vienna, 19.07.2010
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NMS+Ab
IgepalInput virus
NMS NMS+Ab-SCR
D = 9ct unitseq. 3 logs
IN VITRO LYSIS OF FV BY COMPLEMENT IN THE PRESENCE OF THE CONSTRUCTSVienna, 19.07.2010
Patent: WO 2008/135237 A1
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Reduction of the viral titers at about 3logs
105 104 103 102 101
negative control [“no FV“] 1 / / / / /
2 / / / / /
3 / / / / /
positive control [“No Ab, no SCR” ] 4 na na 86 9 5
5 na na 60 11 5
SCR2-Ab#48 8 na na 51 27 6
9 na na 22 5 1
SCR7-Ab#48 13 47 4 / / /
14 na 84 13 2 /
SCR13-Ab#48 15 na 29 / / /
16 73 11 1 / /
SCR18-20-Ab#48 17 61 5 / / /
18 17 1 / / /
INFECTIOUS CENTERS IN THE SPLEENVienna, 19.07.2010
Patent: WO 2008/135237 A1
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SUMMARY I Vienna, 19.07.2010
Trapping of HIV in vivo is mainly dependent on Complement-CR interactions;thus, Complement shows responsible for maintaining the largest viral reservoir in HIV-infected individuals
YCR2
Follicular Dendritic Cells
HIV-C
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antiviral
Activity high
IFN-g
HIV/FV
HIV-C/FV-C
SUMMARY II
Complement is a natural adjuvant for the induction of retrovirus-specific CTLs
Vienna, 19.07.2010
CD8 TC Infected cells
antiviral
Activity low
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retrovirus
C
C‘ CC‘
X
AIM: Reconstitute CML by Pathogen-specific constructsY
constructs
Y
Y Y
Y
YY
Y Y
Putative Targets beside HIV:Other enveloped-Viruses, Bacteria
SUMMARY III Vienna, 19.07.2010
retrovirus
C
C‘ CC‘
Y
Y
YX
X
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ACKNOWLEDGEMENTS
INSTITUTE OF APPLIED MICROBIOLOGY, BOKU, VIENNAG. STIEGLERH. KATINGER
BERNHARD-NOCHT-INSTITUTE, HAMBURGK. TENNER-RACZP. RACZ
DEPARTMENT OF MEDICINE, EPPENDORF, HAMBURGJ. VAN LUNZEN
INSTITUTE OF IMMUNOLOGY, HELSINKIS. JOKIRANTA
ROCKEFELLER UNIVERSITY, NEW YORKR. STEINMAN
GERMAN PRIMATE CENTER, GÖTTINGENC. STAHL-HENNING
INSTITUTE OF MED. VIROLOGY, BOCHUMK. ÜBERLA
INSTITUTE OF VIROLOGY, ZÜRICHA. TRKOLA
INSTITUTE OF VIROLOGY, ESSENU. DITTMER
INSTITUTE OF BIOCHEMISTRY, EPALINGENH. ARCHA-ORBEA
SECTION OF IMMUNOLOGY ,ST. GALLENB. LUDEWIG
RETROVIRAL IMMUNOLOGY SECTION, HAMILTONKIM HASENKRUG RON MESSER
Vienna, 19.07.2010
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OPSONIZED!THINK
THANK YOU !
SECTION OF VIROLOGY, INNSBRUCKDORIS WILFLINGSEDERWILFRIED POSCH
ZOLTAN BANKIASIM EJAZVERENA OBERHAUSERCHRISTOPH AMMANNGEORG HUBERBRIGITTE MÜLLAUERHERIBERT STOIBER
SECTION OF MICROBIOLOGY, INNSBRUCKCORNELIA LASS-FLÖRL
DEPT. OF HAEMATOLOGY, INNSBRUCK GÜNTER GASTL
EU, , MFI
Vienna, 19.07.2010
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