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Persistent infection with mouse hepatitis virus, JHM strain in DBT cell culture
Abstract:
After inoculation with JHM strain into DBT cell monolayers, a persistently infected DBT cell culture was established without producing typical cytopathic changes after about 15th passages. By immunofluorescence virus specific antigen was demonstrated in 10 to 15% DBT cells. This persistently infected culture (JHM-CC) was resistant to superinfection with parental JHM, but such resistance was not shown against vesicular stomatitis virus. JHM-CC virus produced small plaques on DBT cell monolayers. Temperature sensitive (TS) mutant, defective interfering (DI) particle or interferon was not detected in the JHM-CC. To intracerebral inoculation with JHM-CC virus, cortisone treated ICR mice survived without showing clinical signs, however, demyelinating lesions were produced in the brain and spinal cord of them.
Insights
This study established a persistent JHM virus infection in DBT cells, showing resistance to superinfection. The JHM-CC virus caused demyelination in mice brains despite the absence of clinical signs.
Area of Science:
- Virology
- Neuroscience
- Cell Biology
Background:
- Persistent viral infections can alter cellular behavior and immune responses.
- Murine hepatitis virus (JHM strain) is known to cause demyelination.
Purpose of the Study:
- To establish and characterize a persistently infected DBT cell culture with the JHM strain of virus.
- To investigate the biological properties of the persistently infected JHM-CC virus and its effects in a mouse model.
Main Methods:
- DBT cell monolayers were inoculated with the JHM strain, leading to a persistently infected culture (JHM-CC) after passaging.
- Immunofluorescence was used to detect viral antigens.
- Superinfection resistance assays were performed.
- Plaque assays were conducted on DBT cells.
- Temperature sensitivity, defective interfering particles, and interferon production were assessed.
- Intracerebral inoculation into cortisone-treated ICR mice was performed.
Main Results:
- A persistently infected DBT cell culture (JHM-CC) was established, showing viral antigen in 10-15% of cells without cytopathic effects.
- JHM-CC culture exhibited resistance to superinfection with parental JHM virus but not vesicular stomatitis virus.
- JHM-CC virus formed small plaques.
- No temperature-sensitive mutants, defective interfering particles, or interferon were detected.
- Mice inoculated with JHM-CC survived without clinical signs but developed demyelinating lesions in the brain and spinal cord.
Conclusions:
- Persistent JHM virus infection can be established in cell culture, altering viral properties.
- The JHM-CC virus can induce demyelination in the central nervous system of immunosuppressed mice without causing overt disease.