Kondo S, Miyatake S, Iwasaki K, Oda Y, Kikuchi H, Zu Y, Shamoto M, Namba Y
Department of Neurosurgery, Medical School, Kyoto University, Japan.
Neurosurgery. 1992 Apr;30(4):506-11. doi: 10.1227/00006123-199204000-00006.
Three murine monoclonal antibodies, designated GA-17, GB-4, and GC-3, were prepared by the hybridization of murine myeloma cells (NS-1) and spleen cells of BALB/c mice immunized with the crude membrane fraction of cultured human gliosarcoma cells (GI-1). Two of them (GA-17 and GB-4) reacted exclusively with the membrane of glioma cells, and the other (GC-3) reacted with the membrane of glioma cells and a T cell line (MOLT-4). Although these antibodies reacted with almost all of the gliomas, the reactions differed. GA-17 reacted equally well with all glioblastoma (17 cases) and low-grade astrocytoma (10 cases), whereas GB-4 reacted poorly with 7 cases of glioblastoma and GC-3 did not react with 7 cases of low-grade astrocytoma. The antigens, exclusively expressed on the cell surface, were analyzed by surface labeling with 125I followed by a cell lysis and immunoprecipitation with these antibodies. The findings obtained by sodium dodecyl sulfate-polyacrylamide gel electrophoresis revealed that GA-17, GB-4, and GC-3 reacted with Mr 140,000-145,000, Mr 160,000, and Mr 145,000-150,000 proteins, respectively. Some evidence has been obtained indicating that these antigens are composed of the same polypeptide chain (Mr 120,000) with the carbohydrate chains being different.
制备了三种鼠单克隆抗体,分别命名为GA - 17、GB - 4和GC - 3,方法是将鼠骨髓瘤细胞(NS - 1)与用培养的人胶质肉瘤细胞(GI - 1)的粗膜组分免疫的BALB / c小鼠的脾细胞进行杂交。其中两种(GA - 17和GB - 4)仅与胶质瘤细胞膜发生反应,另一种(GC - 3)与胶质瘤细胞膜和一种T细胞系(MOLT - 4)发生反应。尽管这些抗体与几乎所有的胶质瘤都发生反应,但反应情况有所不同。GA - 17与所有胶质母细胞瘤(17例)和低级别星形细胞瘤(10例)反应同样良好,而GB - 4与7例胶质母细胞瘤反应较差,GC - 3与7例低级别星形细胞瘤不发生反应。通过用125I进行表面标记,随后细胞裂解并用这些抗体进行免疫沉淀,对仅在细胞表面表达的抗原进行了分析。十二烷基硫酸钠 - 聚丙烯酰胺凝胶电泳的结果显示,GA - 17、GB - 4和GC - 3分别与分子量为140,000 - 145,000、160,000和145,000 - 150,000的蛋白质发生反应。已获得一些证据表明,这些抗原由相同的多肽链(分子量120,000)组成,但糖链不同。