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Analysis of marek's disease tumor‐associated surface antigen on MDCC‐MSB1‐CLO.18 cells
Author(s) -
Home Masato,
Ohashi Kazuhiko,
Kodama Hiroshi,
Mikami Takeshi
Publication year - 1991
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.2910470212
Subject(s) - antigen , monoclonal antibody , microbiology and biotechnology , antibody , biology , affinity chromatography , chemistry , biochemistry , enzyme , immunology
The physico‐chemical nature of Marek's disease tumor‐associated surface antigen (MATSA) on Marek's disease (MD) lymphoblastoid tumor cell line (MDCC‐MSBI‐clo.18) was examined by the cellular enzyme‐linked immunosorbent assay (CELISA) and the sandwich enzyme‐linked immunosorbent assay (ELISA) using an anti‐MATSA immune serum or a monoclonal antibody (MAb) 2B9 developed against MATSA. Our results indicate that MATSA is a glycoprotein and 2B9 recognizes an antigenic site in the protein moiety of MATSA. MATSA was solubilized from MSB I‐do. 18 cells by treatment with 0.5% Nonidet P‐40, and purified by affinity chromatography coupling with 2B9 and further by ion exchange chromatography on diethylaminoethylcellulose (IECD). MATSA was eluted with 0.2 to 0.3 M KCI in IECD and the purity of MATSA was increased about 2,500‐fold. The purified MATSA was shown to have a molecular weight (M r ) of 70,000 by SDS‐PAGE. The reactivity of purified MATSA with anti‐thymus cell serum was examined. MATSA was detectable by anti‐thymus cell serum, although 2B9, which was used to purify MATSA from MSBI‐clo.18 cells, was not reactive to cells prepared from the thymus. However, MATSA was no longer detectable after the absorption of anti‐thymus cell serum by chicken bursa cells. The absorption of anti‐thymus cell serum by chicken red blood cells (RBC) had no effect on the reactivity against MATSA. These results suggest that MATSA may be a lymphocyte‐specific antigen modified during leukemogenesis by Marek's disease virus (MDV).