A testis-specific antigen of the C57BL/6 mouse.
Spleen cells from female mice of the C57BL/6 strain iso-immunized with an homogenate from 3-week-old mice testis were fused with P3U1 cells. After cloning, two hybridomas, producing IgM, were obtained. Tissue specificity of the monoclonal antibody (Moab) in ascitic fluid was investigated by indirect immunofluorescence. Moab 1A1 reacted specifically with the cytoplasm of spermatogonia, spermatocytes and spermatids, but not with spermatozoa. Testicular antigen recognized by Moab 1A1 (Moab1A1-TA) was prepared by tissue sonication and then subjected to gel filtration. Moab1A1-TA detected in the void volume by ELISA was analyzed by SDS-PAGE and Western blotting. Immuno-staining of membrane filters revealed a broad area within the 45-205 kD range. Moab1A1-TA was treated with proteolytic enzymes, but no changes were observed after Western blotting. Thus, Moab1A1-TA was further digested by peptidases and glycolytic enzymes, electrophoresed using cellulose acetate membranes and immuno-stained with Moab 1A1. Evidence obtained from these experiments strongly suggests that Moab1A1-TA consists of an acidic peptide and a carbohydrate molecule. The antigenicity would be included in the carbohydrate epitope. Moreover, partial digestion of Moab1A1-TA by keratanase indicates that the lacto-series structure is included in the antigenic carbohydrate moiety.
Mikami
O
O
Department of Genetics, Hyogo College of Medicine, Japan.
Sakamoto
H
H
Yamamoto
Y
Y
Furuyama
J
J
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D000941:Antigens / Q000276:immunology / Q000302:isolation & purification*; D000939:Epitopes / Q000276:immunology; D006023:Glycoproteins / Q000276:immunology / Q000302:isolation & purification; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL / Q000276:immunology*; D013737:Testis / Q000276:immunology*
D000911:Antibodies, Monoclonal; D000941:Antigens; D000939:Epitopes; D006023:Glycoproteins