Yeh M Y, Hellström I, Brown J P, Warner G A, Hansen J A, Hellström K E
Proc Natl Acad Sci U S A. 1979 Jun;76(6):2927-31. doi: 10.1073/pnas.76.6.2927.
Mouse NS-1 myeloma cells were fused with spleen cells from mice that had been immunized with cells from a human melanoma, M1804. Hybrid cells were grown in selective medium and tested for production of antibody to surface antigens of M1804 cells. Three hybrids that produced antibodies that bound to the melanoma cells but not to autologous skin fibroblasts were cloned. Antibodies produced by two of the clones were cytotoxic to M1804 cells in the presence of rabbit complement. Extensive specificity tests showed that the antibodies produced by the clones bound strongly only to M1804 cells; significant, although weaker, binding occurred with 2 of 11 allogeneic melanomas. Apart from weak binding of the antibody produced by one of the clones to a breast carcinoma, binding assays of five carcinomas, one sarcoma, and fibroblasts from 17 individuals were negative, as were cytotoxic tests of 10 lymphoblastoid cell lines and peripheral blood lymphocytes from 68 normal donors and 12 chronic lymphocytic leukemia patients. This suggests that we have identified one or more determinants of a melanoma-associated antigen(s), whose expression is limited to a small proportion of melanomas.
将小鼠NS-1骨髓瘤细胞与用人类黑色素瘤M1804细胞免疫过的小鼠脾细胞进行融合。杂交细胞在选择性培养基中培养,并检测其针对M1804细胞表面抗原产生抗体的能力。克隆了三个产生能与黑色素瘤细胞结合但不与自体皮肤成纤维细胞结合的抗体的杂交细胞。在兔补体存在的情况下,其中两个克隆产生的抗体对M1804细胞具有细胞毒性。广泛的特异性测试表明,这些克隆产生的抗体仅与M1804细胞强烈结合;与11种异基因黑色素瘤中的2种有显著结合,尽管较弱。除了其中一个克隆产生的抗体与一种乳腺癌有弱结合外,对5种癌、1种肉瘤以及来自17个人的成纤维细胞的结合试验均为阴性,对10种淋巴母细胞系以及来自68名正常供体和12名慢性淋巴细胞白血病患者的外周血淋巴细胞的细胞毒性测试也均为阴性。这表明我们已经鉴定出一种或多种黑色素瘤相关抗原的决定簇,其表达仅限于一小部分黑色素瘤。