Analysis of Marek's disease tumor-associated surface antigen on MDCC-MSB1-clo.18 cells.
The physico-chemical nature of Marek's disease tumor-associated surface antigen (MATSA) on Marek's disease (MD) lymphoblastoid tumor cell line (MDCC-MSB1-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 MSB1-clo.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 KCl in IECD and the purity of MATSA was increased about 2,500-fold. The purified MATSA was shown to have a molecular weight (Mr) 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 MSB1-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).
Horie
M
M
Department of Epizootiology, Faculty of Veterinary Medicine, Hokkaido University, Sapporo, Japan.
Ohashi
K
K
Kodama
H
H
Mikami
T
T
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D000951:Antigens, Neoplasm / Q000032:analysis* / Q000737:chemistry; D000954:Antigens, Surface / Q000032:analysis* / Q000737:chemistry; D030161:Avian Proteins; D002645:Chickens; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes / Q000276:immunology; D006863:Hydrogen-Ion Concentration; D007106:Immune Sera / Q000276:immunology; D007945:Leukemia, Lymphoid / Q000209:etiology / Q000276:immunology*; D008380:Marek Disease / Q000150:complications / Q000276:immunology*; D008970:Molecular Weight; D009439:Neuraminidase / Q000494:pharmacology; D010504:Periodic Acid / Q000494:pharmacology; D013950:Thymus Gland / Q000276:immunology; D014407:Tumor Cells, Cultured; D014415:Tunicamycin / Q000494:pharmacology
D000911:Antibodies, Monoclonal; D000951:Antigens, Neoplasm; D000954:Antigens, Surface; D030161:Avian Proteins; D000939:Epitopes; D007106:Immune Sera; C066877:Marek's disease tumor-associated surface antigen, avian; D010504:Periodic Acid; D014415:Tunicamycin; C009288:metaperiodate; D009439:Neuraminidase