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Clonal analysis of mammalian cell cultures persistently infected with Japanese encephalitis virus
Abstract:
More than 200 cells were cloned from populations of mammalian cells persistently infected with Japanese encephalitis virus. Only four cloned cultures contained cells that had viral antigen measurable by immunofluorescence and that released infectious virus, yet all clones harbored virus-specific RNA. Superinfection of cloned cells with wild-type Japanese encephalitis virus did not produce cytopathic effects, but resulted in production of viral antigen and infectious virus in formerly nonproducing clones. Cocultivation of nonproducer clone cells with normally permissive cells did not induce virus production, nor did treatment of nonproducer clones with various inhibitors of DNA, RNA, or protein synthesis. It is suggested that the cloning procedure may have selected for a particular subpopulation of cells and that defective virus is also involved in establishment and maintenance of persistent infection.
Insights
Cloning persistently infected mammalian cells revealed that most clones harbored Japanese encephalitis virus RNA but not infectious virus. Superinfection or cell fusion did not induce virus release, suggesting defective virus and cell subpopulations in persistent infections.
Area of Science:
- Virology
- Cell Biology
- Immunology
Background:
- Persistent viral infections can involve complex interactions between viruses and host cells.
- Understanding the mechanisms of persistent Japanese encephalitis virus (JEV) infection is crucial for disease control.
Purpose of the Study:
- To investigate the characteristics of cloned mammalian cell populations persistently infected with JEV.
- To identify factors contributing to the maintenance of persistent JEV infection.
Main Methods:
- Cloning of over 200 cell populations from JEV-infected mammalian cells.
- Detection of viral antigen by immunofluorescence.
- Assay of infectious virus release.
- Detection of virus-specific RNA.
- Superinfection experiments with wild-type JEV.
- Cocultivation of nonproducer cells with permissive cells.
- Treatment with synthesis inhibitors (DNA, RNA, protein).
Main Results:
- Most cloned cultures (over 200) harbored JEV-specific RNA, but only a few released infectious virus or showed viral antigen.
- Superinfection with wild-type JEV induced viral antigen and infectious virus production in previously nonproducing clones.
- Cocultivation or treatment with synthesis inhibitors did not induce virus production in nonproducer clones.
Conclusions:
- The cloning process may select for specific cell subpopulations within persistent JEV infections.
- Defective JEV strains likely play a role in the establishment and maintenance of persistent infections.
- Further research is needed to elucidate the precise mechanisms of JEV persistence.