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Volume 52, Issue 2

Detection of Simian Virus 40 T-Antigen-related Antigens by a I-Protein A-binding Assay and by Immunofluorescence Microscopy on the Surface of SV40-transformed Monolayer Cells Free

Abstract

SUMMARY

Simian virus 40 (SV40)-transformed cells express the SV40-specific tumour transplantation antigen (TSTA) on the cell surface and the SV40-coded tumour antigen in their nuclei. TSTA is defined by SV40-specific transplantation immunity, whereas T-antigen (T-Ag) can be detected serologically by indirect immunofluorescence. Both antigens, however, are derived from the A gene of SV40. We therefore analysed SV40-transformed cells for the presence of serologically detectable T-Ag-related molecules. Such antigens could not be detected on the surface of living SV40-transformed cells in monolayers. However, after a short formaldehyde fixation it was possible to stain the cell surfaces of SV40-transformed cells with sera from rabbits immunized with purified SDS-denatured T-Ag, but not with sera from hamsters bearing SV40-induced tumours. T-Ag-related antigens could be detected with both types of antisera by applying a more sensitive I-protein A assay. The T-Ag specificity of the binding of hamster SV40 tumour sera was demonstrated by a I-IgG-blocking assay in which preincubation of formaldehyde-fixed SV40-transformed cells with rabbit anti-SDS-T-Ag serum inhibited the binding of hamster SV40 tumour serum by about 70%. The localization of T-Ag-related antigens on the outside of plasma membranes of formaldehyde-fixed cells was shown by an anti-SDS-T-Ag serum-specific binding of fluorescein isothiocyanate-labelled to the cell surface. Our results are consistent with the hypothesis that SV40 T-Ag-related antigens are involved in the formation of TSTA.

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© Journal of General Virology 1981
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1981-02-01
2024-04-25
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Detection of Simian Virus 40 T-Antigen-related Antigens by a 125I-Protein A-binding Assay and by Immunofluorescence Microscopy on the Surface of SV40-transformed Monolayer Cells
J. Gen. Virol. 52, 301 (1981); https://doi.org/10.1099/0022-1317-52-2-301
/content/journal/jgv/10.1099/0022-1317-52-2-301
/content/journal/jgv/10.1099/0022-1317-52-2-301
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