体外免疫制备抗大肠癌循环抗原的单克隆抗体

用带结肠癌HRT-18细胞株的BALB/c(nu/nu)小鼠的血清,体处免疫BALB/c小鼠的脾细胞.再与Sp2/0细胞融合,经免疫荧光法在人结肠癌石蜡切片上筛选出一组抗结肠癌的单克隆抗体;A15-6,C13-11,H16-8。间接免疫酶法显示这组单抗对结肠癌的阳性率为69％-72％。免疫组化的特点为:癌巢分泌物及其接触的细胞膜顶端多为阳性反应,其他部位呈阴性反应。3抗体对其他类型的组织无反应。可见,这是一组针对血液循环中肿瘤相关抗原的单克隆抗体,有较好的器官特异性,可能有益于大肠癌的临床血清学检测。

Production of Monoclonal Antibodies against Colorectal Cancer Circulating Antigens by in vitro Immunization

Three monoclonal antibodies, A15-6, C13-11 and H16-8 against human color ectal cancer were produced by fusing spleen cells with murine myeloma cells by in vitro immunization using the sera of athymic mice bearing human HRT-18 colorectal cancer cell as immunogen. The reactivity of the three McAbs with colorectal cancer tissues was examined by indirect immuoperoxidase method. The results indicated that the percentage of positive reactions varied with different McAbs:69% for Al5-6; 77% for C13-11 and 72% for H16-8, respectively. Immunoperoxidase staining showed that the secretory products from cancer cells were most prominently stained and apical membrane was also stained in some glandular structure. However, other parts of the cancer cell membrane were not stained. None of these McAbs reacted with cancerous tissues of the stomach, lung, breast, kidney, skin, bladder and nasopharynx. Moreover, they did not react with benign tumors, inflammatory tissues, fetal colons, colonic polyps, normal colon epithelial cells, blood group substances ABH and other normal tissues. After treatment with NaIO4, the reactivities of colorectal cancer tissues with these three McAbs were destroyed. This suggested that antigenic determinants may contain some carbohydrate structure.