A human monoclonal antibody recognizing a surface antigen on stomach cancer cells.
Lymph-node lymphocytes of a patient with stomach cancer were fused with the mouse-human heterohybridoma, HM-5. A clone (2F9) was isolated that showed stable production of an IgM antibody reactive with NUGC-4 stomach cancer cell line. This antibody reacted predominantly with a cell surface antigen on cell lines originating from gastro-intestinal cancer and adenocarcinoma of lung, whereas it was not generally reactive with other types of cancers, or with normal kidney cells or fibroblasts. Biotin-labeled 2F9 antibody clearly stained cell smears and the nude mouse tumor of NUGC-4, but it did not show a positive reaction with stomach cancer tissues obtained from more than 10 patients, indicating that the antigen detected is very weakly expressed on tumor cells or on a limited number of stomach cancers. The antigen shed from NUGC-4 cell line was detected in the culture supernatant. 2F9 antibody precipitated a glycoprotein with a molecular weight of over 200 kilodaltons as well as a possible glycolipid, from NUGC-4 cells labeled with [3H]glucosamine or [35S]-H2SO4. Periodic acid treatment of the tissue section decreased reactivity with 2F9 antibody, but heat, neuraminidase or protease treatment did not. These results suggested that the epitope is present on a carbohydrate moiety not containing sialic acid, and that a part of the antigen molecule is sulfated.
Yoshikawa
K
K
Department of Microbiology, Aichi Medical University, Japan.
Furukawa
K
K
Ueda
R
R
Iwasa
S
S
Lloyd
K O
KO
Notake
K
K
Takahashi
T
T
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000918:Antibody Specificity; D000951:Antigens, Neoplasm / Q000032:analysis* / Q000276:immunology; D000954:Antigens, Surface / Q000032:analysis* / Q000276:immunology; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes / Q000032:analysis; D005333:Fetus / Q000276:immunology; D005455:Fluorescent Antibody Technique; D006801:Humans; D006825:Hybridomas / Q000276:immunology; D007124:Immunoenzyme Techniques; D007075:Immunoglobulin M / Q000276:immunology; D007163:Immunosorbent Techniques; D008214:Lymphocytes / Q000276:immunology; D008562:Membrane Glycoproteins / Q000276:immunology; D051379:Mice; D008819:Mice, Nude; D008970:Molecular Weight; D009368:Neoplasm Transplantation; D010504:Periodic Acid / Q000494:pharmacology; D013274:Stomach Neoplasms / Q000276:immunology*; D014407:Tumor Cells, Cultured
D000911:Antibodies, Monoclonal; D000951:Antigens, Neoplasm; D000954:Antigens, Surface; D000939:Epitopes; D007075:Immunoglobulin M; D008562:Membrane Glycoproteins; D010504:Periodic Acid