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Infectivity of the DNA from four isolates of JC virus
Abstract:
The infectivity of JC virus DNA was demonstrated in its most permissive cell culture, primary human fetal glial cells. The amount of infectivity observed in these heterogeneous cultures varied considerably between batches of cells. Contrary to results obtained with the papovaviruses simian virus 40 and BK virus, the calcium technique (F. L. Graham and A. J. van der Eb, Virology 52:456--467, 1973) was found to be more efficient at promoting JC virus DNA infectivity than the DEAE-dextran method (J. H. McCutchan and J. S. Pagano, J. Natl. Cancer Inst. 41:351--357, 1968): maximum infectivity titers of 4 x 10-(4) and 6 x 10(3) fluorescent cell units per microgram of DNA, respectively. These values represent an approximate recovery of infectivity from virus of between 0.02 and 0.14%. Comparisons of infectivity of DNAs obtained from four isolates of JC virus and which differed in their degrees of heterogeneity did not reveal significant differences. The JC virus DNA was not infectious in primary human fetal lung and kidney cells.
Insights
JC virus DNA infectivity was confirmed in human fetal glial cells. The calcium technique proved more effective than DEAE-dextran for JC virus DNA transfection, yielding higher infectivity titers.
Area of Science:
- Virology
- Molecular Biology
- Cell Biology
Background:
- JC virus is a human polyomavirus.
- Understanding JC virus DNA infectivity is crucial for studying its pathogenesis.
- Primary human fetal glial cells are the most permissive cells for JC virus infection.
Purpose of the Study:
- To demonstrate the infectivity of JC virus DNA in cell culture.
- To compare the efficiency of different DNA transfection methods for JC virus.
- To assess the infectivity of JC virus DNA from various isolates.
Main Methods:
- JC virus DNA was transfected into primary human fetal glial cells.
- Calcium technique and DEAE-dextran methods were used for transfection.
- Infectivity was measured by fluorescent cell units (FCU).
Main Results:
- JC virus DNA demonstrated infectivity in primary human fetal glial cells, with variable results between cell batches.
- The calcium technique yielded higher infectivity titers (4 x 10^-4 FCU/µg DNA) compared to DEAE-dextran (6 x 10^3 FCU/µg DNA).
- JC virus DNA infectivity was not observed in primary human fetal lung and kidney cells.
Conclusions:
- JC virus DNA is infectious in its most permissive cell type, primary human fetal glial cells.
- The calcium technique is a more efficient method for JC virus DNA transfection than DEAE-dextran.
- JC virus DNA infectivity is specific to certain cell types and does not differ significantly between heterogeneous JC virus DNA isolates.