Antje Ksienzyk Department of Gene Regulation and Differentiation

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Presentation transcript:

Antje Ksienzyk Department of Gene Regulation and Differentiation

Replication in first organ Primary viremia Secondary viremia Replication dissemination other organs model of systemic virus infection Infection

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

The Cre-Loxp system

The used Cre-Loxp system

MCMV- flox and –rec replicate in the same manner

Experimental set-up

10 6 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 EC Alb-cre: Cre under control of albumin promoter in hepatocytes

Hepatocytes are the main producer cell type Hepatocytes derived MCMV-rec do not disseminate from the liver to other organs (alb-cre)

Alb-cre Influence of route and dose Amount of cre expression, the route and dose of infection have no influence on the results

Alb-cre Virus isolation out of liver i.v. injection C57BL/6 Hc-derived MCMV has potential to replicate in spleen and lung

Bidirectional spread of MCMV between EC and Hc EC and Hc produce virus which can infect neighboring cells No explanation that Hc derived virus can’t disseminate Tie2-cre: MCMV from EC to Hc Alb-cre: MCMV from Hc to EC

MCMV infection is controlled by CD3+ cells Elimination of MCMV is correlated with liver infiltrating T-cells Reason for dissemination stop?

Immunosuppression has no influence on Hc- derived virus dissemination 1 untreated 2, 3  -irradiation 4 immune suppression by cyclophosphamide 5 proinflammatory stimuli. Con A No increase of dissemination after immune suppression and proinflammatory stimuli - untreated + immunodepleted

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???