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Generation of Human Monocyte-derived Dendritic Cells from Whole Blood
Published on: December 24, 2016
Unravelling the interaction of human cytomegalovirus with dendritic cells by using SuperSAGE
Martin J Raftery1, Elisabeth Möncke-Buchner, Hideo Matsumura
1Institute of Virology, Charité-Universitätsmedizin, Charitéplatz 1, D-10117 Berlin, Germany. martin.raftery@charite.de
Abstract:
Human cytomegalovirus (HCMV) is a ubiquitous pathogen with a predilection for dendritic cells (DCs). Latently infected myeloid progenitor cells develop into actively infected DCs with impaired functionality, allowing dissemination and transfer of virus throughout the body. However, the viral genes expressed in DCs and their effect on the cellular transcriptome are currently unknown. We investigated human DCs infected with HCMV by using SuperSAGE, allowing us to analyse the transcriptomes of both host and pathogen simultaneously. A small number of viral transcripts were expressed strongly and rapidly post-infection. However, only two were of the immediate-early class, including one with an unknown function. The viral genes expressed reflected the cellular milieu, with the majority having a known or suspected immune-evasion function. Several viral genes identified lack a known function and may fulfil specialized roles within DCs. The cellular response to infection included a strong interferon response, induction of cytokine and anti-apoptotic genes and alterations in genes involved in antigen presentation. We demonstrated the validity of our approach by showing that novel changes first seen in the transcriptome were reflected in the phenotype of HCMV-infected DCs. Delineation of the transcriptional changes underlying the phenotype of HCMV-infected DCs allows a better understanding of how a herpesvirus infects DCs and pinpoints linkages between phenotype and specific viral genes.
Insights
Human cytomegalovirus (HCMV) infects dendritic cells (DCs), impairing their function. This study reveals viral gene expression in DCs and their impact on host cell transcriptomes, offering insights into herpesvirus infection mechanisms.
Area of Science:
- Virology
- Immunology
- Cell Biology
Background:
- Human cytomegalovirus (HCMV) is a common virus that targets dendritic cells (DCs).
- HCMV infection of DCs can impair their immune function, aiding viral spread.
- The specific viral genes active in DCs and their impact on host cell gene expression remain largely unknown.
Purpose of the Study:
- To simultaneously analyze the transcriptomes of HCMV and human DCs.
- To identify viral genes expressed during DC infection and their functional roles.
- To understand the cellular response to HCMV infection in DCs and its link to DC phenotype.
Main Methods:
- Utilized SuperSAGE technology for simultaneous host and viral transcriptome analysis.
- Investigated gene expression profiles in human dendritic cells infected with HCMV.
- Correlated observed transcriptomic changes with the resulting DC phenotype.
Main Results:
- Identified a limited set of strongly expressed viral transcripts, including immediate-early genes.
- Found that most expressed viral genes are involved in immune evasion.
- Observed a significant cellular response, including interferon signaling, cytokine production, and altered antigen presentation genes.
- Several viral genes with unknown functions were identified in infected DCs.
Conclusions:
- HCMV infection profoundly alters the DC transcriptome, influencing immune responses and antigen presentation.
- The study links specific viral gene expression to the altered phenotype of HCMV-infected DCs.
- This research provides a foundation for understanding how HCMV manipulates DCs and evades the immune system.
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