Dual-Sequencing of dendritic cells infected with cytomegalovirus
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ABSTRACT: Dendritic cells were infected with human cytomegalovirus (CMV) for 4, 8 or 12 hours. Single-cultures of dendritic cells serve as controls. RNA was isolated and processed for next generation sequencing.
Project description:Analysis of dendritic cells (DCs) at 6h and 48h after infection with cytomegalovirus (CMV) versus non infected. CMV tends to induce immunosuppression followed by lasting immunity. DCs appear to play a role in this effect. Results provide insights into CMV/DC interactions and suggest mechanisms for CMV-regulated early immune response. Keywords: gene expression array-based, count
Project description:Analysis of dendritic cells (DCs) at 6h and 48h after infection with cytomegalovirus (CMV) versus non infected. CMV tends to induce immunosuppression followed by lasting immunity. DCs appear to play a role in this effect. Results provide insights into CMV/DC interactions and suggest mechanisms for CMV-regulated early immune response. Keywords: gene expression array-based, count In the present study, immature monocyte-derived dendritic cells were generated from CMV neg healthy blood donors and infected or not with an endotheliotropic CMV strain. Gene expression modulation by the CMV infection was studied in a time course assay (non-infected - NI - vs 6h or 48h post-infection. Patient C has no 48h sample
Project description:Human dendritic cells were coinfected with Aspergillus fumigatus and human cytomegalovirus. Single-cultures or single-infections served as controls. RNA was isolated and processed for next generation sequencing.
Project description:Dendritic cells and A. fumigatus were co-cultured with two different MOIs (1 or 0,5) for 2, 4 or 6 hours. Single-cultures of dendritic cells and A. fumigatus serve as controls. RNA was isolated and processed for next generation sequencing.
Project description:Plasmacytoid dendritic cells (pDCs) can rapidly produce interferons and other soluble factors in response to extracellular viruses or virus mimics such as CpG-containing DNA. pDCs can also recognize live cells infected with certain RNA viruses, but the relevance and functional consequences of such recognition remain unclear. We studied the response of primary DCs to the prototypical persistent DNA virus, the human cytomegalovirus (CMV). Human pDCs responded poorly to free CMV but strongly to live CMV-infected fibroblasts, in a process that involved integrin-mediated adhesion, transfer of viral DNA to pDCs and its recognition through TLR9. Compared to transient polyfunctional responses to CpG or free influenza virus, pDC response to CMV-infected cells was long-lasting, dominated by the production of type I (IFN-I) and type III (IFN-III) interferons, and lacked diversification into functionally distinct populations. Similarly, pDC activation by influenza-infected lung epithelial cells was highly efficient, prolonged and dominated by interferon production. Prolonged pDC activation by CMV-infected cells facilitated the activation of natural killer cells that are critical for CMV control. Finally, patients with CMV viremia harbored phenotypically activated pDCs and increased levels of IFN-I and IFN-III in circulation. Thus, recognition of live infected cells is a common mechanism of virus detection by pDCs that elicits a unique antiviral response program.
Project description:Dendritic cells (DC) play a crucial role in generating and maintaining antiviral immunity. While DC are implicated in the antiviral defense by inducing T cell responses, they can also become infected by Cytomegalovirus (CMV). CMV is not only highly species-specific but also specialized in evading immune protection, and this specialization is in part due to characteristic genes encoded by a given virus. Here, we investigated whether RCMV can infect XCR1+ DC and if infection of DC alters the expression of cell surface markers and migration behavior. We demonstrate that wild-type RCMV and a _vxcl1 mutant virus infect splenic rat DC ex vivo and identify viral assembly compartments. Replication-competent RCMV reduced XCR1 and MHCII surface expression. Further, gene expression of infected DC was analyzed by single cell RNA-sequencing. RCMV infection reverted a state of DC activation that was induced by DC cultivation. On the functional level, we observed impaired chemotactic activity of infected XCR1+ DC compared to mock treated cells. We therefore speculate that as a result of RCMV infection, DC exhibit diminished XCR1 expression and are thereby inhibited from the lymphocyte crosstalk.
Project description:Analysis of microRNA expression patterns in cytomegalovirus infected human fibroblasts for two cultures with differing sensitivity to the virus Non-coding RNAs were cloned from total RNA extracts obtained from 2 human fibroblast lines with differing sensitivity to CMV infection. For each line 3 types of samples are compared: pre-experimantal norm, 3 hours after infection, 3 hours without infection.
Project description:To study the gene expression dynamics in cytomegalovirus (CMV) DNAemic patients after kidney transplantation, we performed RNA sequencing (RNAseq) on 31 DNAemic patients at 4 timepoints (baseline, 1 week post-DNAemia, 1 month post-DNAemia, and a long-term timepoint 9-18 months post-DNAemia) matched with 31 non-DNAemic patients at 2 timepoints (baseline and long-term).