Date post: | 05-Jan-2016 |
Category: |
Documents |
Upload: | helena-amberlynn-small |
View: | 214 times |
Download: | 0 times |
Virologie
H. STOIBERVIENNA, 19.07.2010
MULTIFACETED INTERACTIONS OF RETROVIRUSES WITH THE COMPLEMENT
SYSTEM
SCHEMATIC OVERVIEW OF IMMUNE RESPONSES UPON VIRAL INFECTIONSVienna, 19.07.2010
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
(Schacker et al. 2000)
EX VIVO DETECTION OF HIV IN LT OF INFECTED PATIENTS Vienna, 19.07.2010
COMPLEMENT-DEPENDENT TRAPPING OF HIV IN THE LT
Kacani et al. JV
Vienna, 19.07.2010
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
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
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
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
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.
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;
%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
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
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
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
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
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
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
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
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
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
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
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
Österreichischen Förderungsgesellschaften FFG