Related Experiment Videos
A model for persistent murine coronavirus infection involving maintenance via cytopathically infected cell centres
G Macintyre1, F Wong, R Anderson
1Department of Microbiology and Infectious Diseases, University of Calgary, Alberta, Canada.
Abstract:
The relatively cell impermeable hygromycin B was found to inhibit viral but not cellular protein synthesis when added to cultures of murine hepatitis virus (MHV)-infected or mock-infected mouse L-2 fibroblasts. Membrane permeability, as judged by influx of sodium ions, has previously been demonstrated to be an MHV E2 glycoprotein-mediated, cytopathic effect of MHV infection in L-2 cells. It is therefore likely that the selective effect of hygromycin B on viral protein synthesis is a reflection of an increased drug penetration into virus-infected cells. Using hygromycin B as a marker for MHV-induced cell membrane cytopathology, the effects of drug treatment on a persistent MHV infection in mouse LM-K fibroblasts were investigated. MHV persistence in LM-K cells, which normally involves a steady state infection of 0.1 to 1% of the cells in culture, was found to be cured by hygromycin B treatment, as measured by the elimination of infectious virus from the supernatant medium. Hygromycin B also resulted in the eradication of MHV-specific RNA from LM-K cells, arguing against the presence of a non-cytopathically or latently infected subpopulation of cells.
Insights
Hygromycin B selectively inhibits viral protein synthesis in murine hepatitis virus (MHV)-infected cells. This drug effectively cured persistent MHV infections by eliminating both the virus and its RNA from cell cultures.
Area of Science:
- Virology
- Cell Biology
- Drug Discovery
Background:
- Murine hepatitis virus (MHV) infection causes cytopathic effects in L-2 fibroblasts, including increased membrane permeability.
- MHV infection selectively inhibits cellular protein synthesis, while viral protein synthesis remains unaffected.
Purpose of the Study:
- To investigate the selective inhibition of viral protein synthesis by hygromycin B in MHV-infected cells.
- To evaluate hygromycin B as a potential therapeutic agent for persistent MHV infections.
- To use hygromycin B as a marker for MHV-induced cell membrane cytopathology.
Main Methods:
- Treatment of MHV-infected mouse L-2 fibroblasts with hygromycin B.
- Assessment of protein synthesis inhibition in viral and cellular components.
- Investigation of hygromycin B's effect on persistent MHV infection in mouse LM-K fibroblasts.
- Measurement of infectious virus and MHV-specific RNA elimination.
Main Results:
- Hygromycin B inhibited viral protein synthesis but not cellular protein synthesis in MHV-infected cells.
- Increased membrane permeability in infected cells likely enhances hygromycin B penetration.
- Hygromycin B treatment cured persistent MHV infection in LM-K cells, eliminating infectious virus and MHV-specific RNA.
- Evidence suggests no non-cytopathic or latent infected cell subpopulation.
Conclusions:
- Hygromycin B demonstrates selective antiviral activity against MHV.
- Hygromycin B can effectively eradicate persistent MHV infections.
- The drug's efficacy is linked to MHV-induced membrane alterations, highlighting a potential therapeutic strategy.