Analysis of simian virus 40 infection of CV‐1 cells by quantitative two‐color fluorescence with flow cytometry

Wiley - Tập 9 Số 1 - Trang 52-59 - 1988
John M. Lehman1, Judith Laffin1, James W. Jacobberger1, David Fogleman1
1Department of Microbiology and Immunology, Albany Medical College, Albany, New York 12208

Tóm tắt

AbstractQuantitative two‐color fluorescent analysis of Simian virus (SV40) infection of permissive CV‐1 cells was investigated. Analysis included by quantitation of cellular DNA, the early viral tumor (T) antigen with a monoclonal antibody, and late viral (V) antigens with a polyclonal antibody. T antigen was detected in all phases of the cell cycle at 6 and 12 h, after SV40 infection of growth arrested cells. At later time intervals, the percentage of T‐antigen‐positive cells increased with the induction of the cells into successive rounds of DNA synthesis and an increase in tetraploidpolyploid cells. The amount of T antigen per cell increased as the cells entered the successive stages of the cell cycle (G0/G1 → G2 + M → tetraploid S and G2 + M). The V antigen from adsorbed virus was detected immediately after infection. Synthesis of V antigen began in late S and G2 + M phases of the cell cycle. This quantitative analysis allows a definitive determination of antigen per cell in a population correlated with the cell cycle and may be useful in correlating viral and cellular events with transformation.

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Wiley

Tập 9 Số 1

52-59

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