Letter | Published:

Simian virus 40 specific proteins on surface of HeLa cells infected with adenovirus 2–SV40 hybrid virus Ad2+ND2

Naturevolume 277pages322324 (1979) | Download Citation

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Abstract

CELLS infected or transformed by simian virus 40 (SV40) express SV40 tumour-specific transplantation antigen (TSTA)1,2. The mechanism of tumour rejection mediated by this antigen is not yet understood, but our knowledge about transplantation rejection suggests that TSTA should be located on the cell surface3. SV40 TSTA has a close molecular relationship to SV40 T-antigen4,5, which is a nuclear antigen, and until now, SV40 specific proteins have not been demonstrated unequivocally in the plasma membranes or on the cell surfaces of SV40 infected or transformed cells. In contrast, in cells infected by the nondefective adenovirus 2–SV40 hybrid virus Ad2+ND2, a virus known to induce SV40 TSTA6 the SV40-specific proteins (MW 56,000 (56K) and 42,000 (42K)) are stably incorporated into the plasma membranes7. These proteins are coded for—at least in part—by the SV40 portion of the hybrid virus genome and correspond to the C-terminal half of SV40 T-antigen8. Furthermore, these proteins are immunoprecipitable from extracts of Ad2+ND2–infected cells using serum from SV40 tumour-bearing hamsters7. Analysis of Ad2+ND2–infected HeLa cells by immunofluorescence microscopy for cell surface fluorescence using hamster SV40 tumour serum, however, failed to give a positive result (W.D., unpublished). A possible explanation is that upon location on the cell surface the structure of these proteins is altered so that they are no longer recognised by the tumour sera. These sera are probably directed against native T-antigen released from the nuclei of necrotic tumour material. We therefore decided to prepare antisera against SDS-denatured T-antigen which are directed against the primary structure of T-antigen rather than against its native conformation. We report here that SV40-specific proteins can be visualised on the cell surface of Ad2+ND2–infected HeLa cells by immunofluorescence microscopy using a rabbit anti-T serum, prepared against purified SDS-denatured T-antigen.

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  1. Max Planck Institute for Biophysical Chemisty, PO Box 968, D-3400, Göttingen, FRG

    • WOLFGANG DEPPERT
    •  & ROBERT PATES

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https://doi.org/10.1038/277322a0

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