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

Abstract

Quantitative 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.