Simian virus 40 T-antigen-related cell surface antigen: serological demonstration on simian virus 40-transformed monolayer cells in situ.
Simian virus 40 (SV40)-transformed monolayer cells were analyzed in situ by indirect immunofluorescence microscopy for the postulated cell surface location of SV40 T-antigen-related molecules. With antisera prepared against purified, sodium dodecyl sulfate-denatured SV40 T-antigen, positive surface staining was obtained when the cells had been treated with formaldehyde before immunofluorescence analysis. In contrast, living SV40-transformed cells analyzed in monolayer were surface fluorescence negative. The fixation procedure developed in this study combined with a double staining immunofluorescence technique allowed the simultaneous analysis of the same cells for the expression of both SV40 T-antigen-related surface antigen and nuclear T-antigen. The localization of SV40 T-antigen-related surface antigen on the outer surface of the plasma membrane of formaldehyde-fixed SV40-transformed cells was demonstrated directly by the protein A-mediated binding of Staphylococcus aureus bacteria on formaldehyde-fixed SV40-transformed cells precoated with antiserum against sodium dodecyl sulfate-denatured T-antigen. Both cell surface staining and S. aureus binding were found to be highly specific for SV40 T-antigen-related binding sites. These results indicate that T-antigen-related molecules in a cryptic form are located on the surface of SV40-transformed monolayer cells and can be detected in situ after modification of the cell surface architecture.
D000818:Animals; D000951:Antigens, Neoplasm / Q000032:analysis*; D000954:Antigens, Surface / Q000276:immunology*; D000956:Antigens, Viral / Q000032:analysis*; D000957:Antigens, Viral, Tumor; D002472:Cell Transformation, Viral; D002478:Cells, Cultured; D002522:Chlorocebus aethiops; D006224:Cricetinae; D004622:Embryo, Mammalian; D005347:Fibroblasts / Q000276:immunology; D005455:Fluorescent Antibody Technique; D006367:HeLa Cells; D006801:Humans; D007106:Immune Sera / Q000276:immunology; D007668:Kidney; D051379:Mice; D008807:Mice, Inbred BALB C; D008855:Microscopy, Electron, Scanning; D011817:Rabbits; D051381:Rats; D013539:Simian virus 40 / Q000276:immunology; D013211:Staphylococcus aureus
D000951:Antigens, Neoplasm; D000954:Antigens, Surface; D000956:Antigens, Viral; D000957:Antigens, Viral, Tumor; D007106:Immune Sera