Antje Ksienzyk Department of Gene Regulation and Differentiation [email protected].
-
Upload
gillian-andrews -
Category
Documents
-
view
222 -
download
0
Transcript of Antje Ksienzyk Department of Gene Regulation and Differentiation [email protected].
![Page 2: Antje Ksienzyk Department of Gene Regulation and Differentiation aks08@helmholtz-hzi.de.](https://reader036.fdocuments.in/reader036/viewer/2022062309/56649f2a5503460f94c43aa2/html5/thumbnails/2.jpg)
Replication in first organPrimary viremia
Secondary viremia
Replication
dissemination
other organs
model of systemic virus infection
Infection
![Page 3: Antje Ksienzyk Department of Gene Regulation and Differentiation aks08@helmholtz-hzi.de.](https://reader036.fdocuments.in/reader036/viewer/2022062309/56649f2a5503460f94c43aa2/html5/thumbnails/3.jpg)
MCMV
-Herpesviridae herpesvirus
-Cytomegalovirus-MCMV: mouse cytomegalovirus
HCMV :major risk factor for immunoincompetent and immunocompromised patients
(AIDS patients and organ transplant recipients)
MCMV: represents the rodent model
MCMV replicates in a broad spectrum of cell types: vascular endothelial cells and
hepatocytes
![Page 4: Antje Ksienzyk Department of Gene Regulation and Differentiation aks08@helmholtz-hzi.de.](https://reader036.fdocuments.in/reader036/viewer/2022062309/56649f2a5503460f94c43aa2/html5/thumbnails/4.jpg)
The Cre-Loxp system
![Page 5: Antje Ksienzyk Department of Gene Regulation and Differentiation aks08@helmholtz-hzi.de.](https://reader036.fdocuments.in/reader036/viewer/2022062309/56649f2a5503460f94c43aa2/html5/thumbnails/5.jpg)
The used Cre-Loxp system
![Page 6: Antje Ksienzyk Department of Gene Regulation and Differentiation aks08@helmholtz-hzi.de.](https://reader036.fdocuments.in/reader036/viewer/2022062309/56649f2a5503460f94c43aa2/html5/thumbnails/6.jpg)
MCMV- flox and –rec replicate in the same manner
![Page 7: Antje Ksienzyk Department of Gene Regulation and Differentiation aks08@helmholtz-hzi.de.](https://reader036.fdocuments.in/reader036/viewer/2022062309/56649f2a5503460f94c43aa2/html5/thumbnails/7.jpg)
Experimental set-up
![Page 8: Antje Ksienzyk Department of Gene Regulation and Differentiation aks08@helmholtz-hzi.de.](https://reader036.fdocuments.in/reader036/viewer/2022062309/56649f2a5503460f94c43aa2/html5/thumbnails/8.jpg)
106 PFU of MCMV flox
Analyses of MCMV-rec and MCMV-flox plaque forming units
EC and HC are important cell types for MCMV replication
Tie2-cre: Cre under control of Tie2 (endothelial tyrosinkinase receptors: only in EC) promoter in vascular ECAlb-cre: Cre under control of albumin promoter in hepatocytes
![Page 9: Antje Ksienzyk Department of Gene Regulation and Differentiation aks08@helmholtz-hzi.de.](https://reader036.fdocuments.in/reader036/viewer/2022062309/56649f2a5503460f94c43aa2/html5/thumbnails/9.jpg)
Hepatocytes are the main producer cell type
Hepatocytes derived MCMV-rec do not disseminate from the liver to other organs (alb-cre)
![Page 10: Antje Ksienzyk Department of Gene Regulation and Differentiation aks08@helmholtz-hzi.de.](https://reader036.fdocuments.in/reader036/viewer/2022062309/56649f2a5503460f94c43aa2/html5/thumbnails/10.jpg)
Alb-cre
Influence of route and dose
Amount of cre expression, the route and dose of infection have no influence on the results
![Page 11: Antje Ksienzyk Department of Gene Regulation and Differentiation aks08@helmholtz-hzi.de.](https://reader036.fdocuments.in/reader036/viewer/2022062309/56649f2a5503460f94c43aa2/html5/thumbnails/11.jpg)
Alb-cre
Virus isolation out of liver
i.v. injection
C57BL/6
Hc-derived MCMV has potential to replicate in spleen and lung
![Page 12: Antje Ksienzyk Department of Gene Regulation and Differentiation aks08@helmholtz-hzi.de.](https://reader036.fdocuments.in/reader036/viewer/2022062309/56649f2a5503460f94c43aa2/html5/thumbnails/12.jpg)
Bidirectional spread of MCMV between EC and Hc
EC and Hc produce virus which can infect neighboring cellsNo explanation that Hc derived virus can’t disseminate
Tie2-cre: MCMV from EC to HcAlb-cre: MCMV from Hc to EC
![Page 13: Antje Ksienzyk Department of Gene Regulation and Differentiation aks08@helmholtz-hzi.de.](https://reader036.fdocuments.in/reader036/viewer/2022062309/56649f2a5503460f94c43aa2/html5/thumbnails/13.jpg)
MCMV infection is controlled by CD3+ cells
Elimination of MCMV is correlated with liver infiltrating T-cellsReason for dissemination stop?
![Page 14: Antje Ksienzyk Department of Gene Regulation and Differentiation aks08@helmholtz-hzi.de.](https://reader036.fdocuments.in/reader036/viewer/2022062309/56649f2a5503460f94c43aa2/html5/thumbnails/14.jpg)
Immunosuppression has no influence on Hc- derived virus dissemination
1 untreated2, 3 -irradiation4 immune suppression by cyclophosphamide 5 proinflammatory stimuli. Con A
No increase of dissemination after immune suppression and proinflammatory stimuli
- untreated+ immunodepleted
![Page 15: Antje Ksienzyk Department of Gene Regulation and Differentiation aks08@helmholtz-hzi.de.](https://reader036.fdocuments.in/reader036/viewer/2022062309/56649f2a5503460f94c43aa2/html5/thumbnails/15.jpg)
summary
- Hepatocytes and EC are target cells of MCMV in vivo (replication)
- HC are important target cells but are not involved in virus dissemination
-Infected EC contribute to viremic dissemination by serving as transport vehicle and
by uptake and release of virus
- Hc derived MCMV never leads to a second viremia (unlike to former publications)
-But what is the reason for that?
-Independent on immune cells or irradiation
-Independent on ability to infect other cells
- IFN???