Analysis of simian virus 40 infection of CV-1 cells by quantitative two-color fluorescence with flow cytometry.
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 tetraploid-polyploid 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.
Lehman
J M
JM
Department of Microbiology and Immunology, Albany Medical College, New York 12208.
Laffin
J
J
Jacobberger
J W
JW
Fogleman
D
D
D000818:Animals; D000957:Antigens, Viral, Tumor / Q000096:biosynthesis; D002453:Cell Cycle; D002460:Cell Line; D002522:Chlorocebus aethiops; D004279:DNA, Viral / Q000032:analysis; D005434:Flow Cytometry / Q000379:methods*; D005455:Fluorescent Antibody Technique; D007668:Kidney / Q000166:cytology* / Q000382:microbiology; D008722:Methods; D013539:Simian virus 40; D014412:Tumor Virus Infections / Q000378:metabolism*
D000957:Antigens, Viral, Tumor; D004279:DNA, Viral