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Specific glycolipid antigen(s) in SV40‐transformed cell membranes
Author(s) -
Ansel Sandra,
Huet Christian
Publication year - 1980
Publication title -
international journal of cancer
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.475
H-Index - 234
eISSN - 1097-0215
pISSN - 0020-7136
DOI - 10.1002/ijc.2910250616
Subject(s) - glycolipid , trypsinization , microbiology and biotechnology , hamster , antigen , biology , cell culture , staining , membrane , chinese hamster ovary cell , cell , chemistry , biochemistry , trypsin , immunology , genetics , enzyme
Abstract A glycolipid extract was prepared from an SV40‐transformed 1 hamster cell line (EH‐SV) according to the Folch partition procedure. The glycolipids from the aqueous layer were incorporated in liposomal membranes composed of lecithin/sphingomyelin/cholesterol (1:1:2 by weight). This liposomal preparation was inoculated in Syrian hamsters to raise immune sera. The sera were absorbed with trypsinized «normal» hamster cells (EH‐N) and tested on various cell lines by the indirect immunofluorescence technique. When used for staining living cells, the immune serum produced a distinct cellsurface fluorescence with SV40‐transformed cell lines regardless of the cell origin ( e.g. , rat or hamster). No reaction was observed with heterologous Py‐transformed cell lines, spontaneously transformed cells, or sera from non‐immunized hamsters. When used for staining acetone‐fixed cells, the antiglycolipid serum reacted specifically with a thermostable antigen in the nuclear envelope and the cytoplasm of SV40‐transformed cells. The sera lack interfering SV40 T reactivity. The results indicate the presence of related SV40‐specific glycolipid antigen(s) in the plasma membrane, the nuclear membrane and probably other endomembranes of SV40‐transformed cells.