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Characterization of JC papovavirus adapted to growth in human embryonic kidney cells
Abstract:
Human papovavirus JC virus was adapted to growth in human embryonic kidney (HEK) cells. After eight passages, the HEK-adapted JC virus produced high virus yields and was capable of forming plaques in HEK monolayer cultures. Eleven plaque-purified stocks were prepared and characterized. Biologically, the plaque-purified virus induced tumor and viral antigens in HEK cells earlier and in a higher percentage of cells than uncloned virus. Cytopathic changes were also evident sooner and were more extensive. The DNA from uncloned as well as plaque-purified isolates was analyzed by restriction endonuclease cleavage followed by gel electrophoresis. The DNA from uncloned HEK-adapted virus was heterogeneous. Plaque-purified virus isolates yielded DNA which, although much less heterogeneous than the uncloned stock, still consisted of two or more species of viral DNA.
Insights
Human embryonic kidney (HEK) cell-adapted JC virus demonstrated enhanced infectivity and replication. Plaque-purified JC virus isolates showed improved biological activity and viral DNA heterogeneity, indicating genetic adaptation.
Area of Science:
- Virology
- Molecular Biology
- Cell Biology
Background:
- JC virus (JCV), a human polyomavirus, is known to cause progressive multifocal leukoencephalopathy (PML) in immunocompromised individuals.
- Adapting JCV to grow in specific cell lines can alter its biological properties and facilitate research.
Purpose of the Study:
- To adapt human JC virus (JCV) for efficient growth in human embryonic kidney (HEK) cells.
- To characterize the biological and genetic properties of HEK-adapted JCV isolates.
Main Methods:
- JC virus was serially passaged in HEK cells to achieve adaptation.
- Plaque purification was performed to isolate distinct viral variants.
- Biological activity was assessed by measuring viral antigen production, tumor induction in HEK cells, and cytopathic effects.
- Viral DNA was analyzed using restriction endonuclease digestion and gel electrophoresis.
Main Results:
- HEK-adapted JCV achieved high virus yields and plaque-forming ability after eight passages.
- Plaque-purified isolates exhibited earlier and more pronounced induction of viral antigens and tumors in HEK cells compared to uncloned virus.
- Analysis of viral DNA revealed heterogeneity in the uncloned HEK-adapted stock, with plaque-purified isolates showing reduced, but still present, heterogeneity (two or more DNA species).
Conclusions:
- JC virus can be adapted to efficiently replicate in human embryonic kidney cells.
- Adaptation and subsequent plaque purification result in viral variants with enhanced biological activity and altered, though still heterogeneous, viral DNA profiles.