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Retroviral Infection of Murine Embryonic Stem Cell Derived Embryoid Body Cells for Analysis of Hematopoietic Differentiation
Published on: October 20, 2014
Permissive and restricted virus infection of murine embryonic stem cells
Rachael Wash1, Sabrina Calabressi1, Stephanie Franz1
1Wellcome Trust Sanger Institute, Wellcome Trust Genome Campus, Hinxton, Cambridge, CB10 1SA, UK.
Abstract:
Recent RNA interference (RNAi) studies have identified many host proteins that modulate virus infection, but small interfering RNA 'off-target' effects and the use of transformed cell lines limit their conclusiveness. As murine embryonic stem (mES) cells can be genetically modified and resources exist where many and eventually all known mouse genes are insertionally inactivated, it was reasoned that mES cells would provide a useful alternative to RNAi screens. Beyond allowing investigation of host-pathogen interactions in vitro, mES cells have the potential to differentiate into other primary cell types, as well as being used to generate knockout mice for in vivo studies. However, mES cells are poorly characterized for virus infection. To investigate whether ES cells can be used to explore host-virus interactions, this study characterized the responses of mES cells following infection by herpes simplex virus type 1 (HSV-1) and influenza A virus. HSV-1 replicated lytically in mES cells, although mES cells were less permissive than most other cell types tested. Influenza virus was able to enter mES cells and express some viral proteins, but the replication cycle was incomplete and no infectious virus was produced. Knockdown of the host protein AHCYL1 in mES cells reduced HSV-1 replication, showing the potential for using mES cells to study host-virus interactions. Transcriptional profiling, however, indicated the lack of an efficient innate immune response in these cells. mES cells may thus be useful to identify host proteins that play a role in virus replication, but they are not suitable to determine factors that are involved in innate host defence.
Insights
Murine embryonic stem (mES) cells can identify host proteins affecting virus replication, but their innate immune response is weak. These cells offer a new tool for studying virus-host interactions, complementing RNA interference (RNAi) methods.
Area of Science:
- Virology
- Stem Cell Biology
- Host-Pathogen Interactions
Background:
- RNA interference (RNAi) studies face limitations due to off-target effects and transformed cell lines.
- Murine embryonic stem (mES) cells offer genetic manipulability and access to gene-knockout resources, presenting an alternative to RNAi screens.
- mES cells are poorly characterized for their response to viral infections.
Purpose of the Study:
- To characterize the response of mES cells to herpes simplex virus type 1 (HSV-1) and influenza A virus infections.
- To evaluate the potential of mES cells for studying host-virus interactions and identifying host factors.
- To assess the suitability of mES cells for investigating innate host defense mechanisms.
Main Methods:
- Infection of mES cells with HSV-1 and influenza A virus.
- Monitoring viral replication and protein expression.
- Genetic knockdown of host protein AHCYL1 in mES cells.
- Transcriptional profiling to analyze innate immune responses.
Main Results:
- HSV-1 replicated lytically in mES cells, albeit with lower permissivity compared to other cell types.
- Influenza A virus entered mES cells and expressed viral proteins, but replication was incomplete, yielding no infectious virus.
- Knockdown of AHCYL1 reduced HSV-1 replication, demonstrating mES cells' utility for identifying host factors.
- Transcriptional profiling revealed an inefficient innate immune response in mES cells.
Conclusions:
- mES cells are a valuable tool for identifying host proteins that influence viral replication.
- mES cells are not suitable for studying factors involved in innate host defense due to their limited immune response.
- mES cells provide a complementary approach to RNAi for host-pathogen interaction studies.
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