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Dissecting Host-virus Interaction in Lytic Replication of a Model Herpesvirus
Published on: October 7, 2011
Interactions of murine gammaherpesvirus 68 with B and T cell lines
N P Sunil-Chandra1, S Efstathiou, A A Nash
1Department of Pathology, University of Cambridge, United Kingdom.
Abstract:
Murine gammaherpesvirus is a natural pathogen of wild rodents. We have established that in vivo the virus persists in B lymphocytes in a latent form and therefore has similar biological properties to Epstein-Barr virus and related gamma-I-herpesviruses. In this report we have established a persistent infection in mouse myeloma (B) cells (NSO cell line), but not in mouse thymoma (T) cells (BW 5147 cell line). The virus persists indefinitely in myeloma cells, without any apparent cytopathic effect, but with the production of infectious virus. We demonstrate that ACV abolishes the productive infection, but large numbers of cells harbor the virus in a latent form, as determined by an infectious center assay. Analysis of the viral DNA has shown that during a persistent infection linear virus genomes predominated, with low levels of circular DNA also present. Treatment of cells with ACV results in a significant reduction of linear genomes, but has no effect on the level of circular DNA molecules. These data provide further evidence to support our earlier observations on B cells as the site of latency and provides an in vitro model with which to study the molecular basis of MHV-68 latency/persistence.
Insights
Murine gammaherpesvirus (MHV-68) establishes persistent infections in mouse myeloma cells, not T cells. This provides an in vitro model for studying MHV-68 latency and persistence in B lymphocytes.
Area of Science:
- Virology
- Immunology
- Cell Biology
Background:
- Murine gammaherpesvirus (MHV-68) is a natural pathogen of wild rodents.
- In vivo, MHV-68 establishes a latent infection primarily within B lymphocytes, similar to Epstein-Barr virus.
- Understanding the mechanisms of gammaherpesvirus latency is crucial for developing antiviral strategies.
Purpose of the Study:
- To establish an in vitro model for studying MHV-68 latency and persistence.
- To investigate the cellular tropism of MHV-68 in vitro.
- To analyze the impact of antiviral treatment on viral DNA forms during persistent infection.
Main Methods:
- Establishing persistent MHV-68 infection in mouse myeloma (NSO) and thymoma (BW 5147) cell lines.
- Assessing viral persistence and production using infectious center assays.
- Analyzing viral DNA forms (linear and circular genomes) using molecular techniques.
- Treating infected cells with acyclovir (ACV) to evaluate its effect on productive infection and viral DNA.
Main Results:
- MHV-68 established a persistent infection in mouse myeloma (B) cells but not in thymoma (T) cells.
- The virus persisted indefinitely in myeloma cells without significant cytopathic effect, producing infectious virions.
- Acyclovir (ACV) treatment abolished productive infection, but latent viral genomes remained detectable.
- During persistent infection, linear viral genomes predominated, with low levels of circular DNA; ACV reduced linear genomes but not circular DNA.
Conclusions:
- Mouse myeloma cells provide a suitable in vitro model for studying MHV-68 latency.
- B lymphocytes are confirmed as the primary site for MHV-68 latency.
- The differential effect of ACV on linear and circular viral DNA suggests distinct roles in viral persistence.
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