Y and blood group B type 2 glycolipid antigens accumulate in a human gastric carcinoma cell line as detected by monoclonal antibody. Isolation and characterization by mass spectrometry and NMR spectroscopy

• Published in: The Journal of biological chemistry 1987-01, Vol.262 (1), p.372-379
• Main Authors: Blaszczyk-Thurin, M, Thurin, J, Hindsgaul, O, Karlsson, K A, Steplewski, Z, Koprowski, H
• Format: Article
• Language: English
• Subjects: ABO Blood-Group System - immunology; Adenocarcinoma - immunology; Animals; Antibodies, Monoclonal - immunology; Antibody Specificity; Antigens, Neoplasm - immunology; Antigens, Neoplasm - isolation & purification; Biological and medical sciences; Cell Line; Cell physiology; Cell transformation and carcinogenesis. Action of oncogenes and antioncogenes; Chemical Phenomena; Chemistry; Epitopes - immunology; Fucose; Fundamental and applied biological sciences. Psychology; Glycolipids - immunology; Humans; Magnetic Resonance Spectroscopy; Mass Spectrometry; Mice; Molecular and cellular biology; Stomach Neoplasms - immunology
• ISSN: 0021-9258; 1083-351X
• DOI: 10.1016/S0021-9258(19)75937-7

A monoclonal antibody (mAb), BR55-2, was generated from mice immunized with MCF-7 human breast carcinoma cells. This mAb specifically detected glycolipids with the Y determinant Fuc alpha 1---2Gal beta 1---4GlcNAc(3---1 alpha Fuc)-beta 1---3Gal beta 1---4Glc beta 1---1 Cer and the Y-related B-active difucosylated determinant Gal alpha 1---3Gal(2---1 alpha Fuc) beta 1---4GlcNAc(3---1 alpha Fuc) beta 1---3Gal beta 1---4Glc beta 1---1 Cer, but was not reactive with related monofucosylated glycolipids of type 2 chain (X-antigen, blood group H), type 1 chain (Lea antigen, blood group H and B) or with difucosylated type 2 and type 1 chain structures (A blood group antigen or blood group B and Leb, respectively). A series of glycolipids with Y and blood group B type 2 determinants were detected in human gastric adenocarcinoma cell line KATO III with mAb BR55-2 and with a previously characterized anti-blood group B mAb PA83-52 (Hansson, G. C., Karlsson, K.-A., Larson, G., McKibbin, J. M., Blaszczyk, M., Herlyn, M., Steplewski, Z., and Koprowski, H. (1983) J. Biol. Chem. 258, 4091-4097). The isolated antigens were structurally characterized by mass spectrometry of permethylated and permethylated-reduced derivatives and by proton NMR spectroscopy. In a chromatogram binding assay, mAb BR55-2 and mAb PA83-52 detected minor components with slower mobility than the Y-6 and blood group B-7-type 2 structures. The detection of a B type 2 determinant is the first chemical evidence for the presence of an autologous difucosyl blood group B type 2 antigen in human adenocarcinoma cells.