Cui H, Qian H N, Feng J, Fu T Y, Wei P, Fu Z Y
Gynecologic Oncology Research Laboratory, People's Hospital, Beijing Medical University.
Chin Med J (Engl). 1990 Jun;103(6):478-84.
Lymphocytes from regional lymph nodes of patients with ovarian carcinoma were immortalized by fusing them with a nonsecreting cell line of murine myeloma (Sp2/0-Ag14). The fusion rate was 0-87.5%. By early cloning and recloning, a hybrid cell line, named HMD4, was established. It has stably secreted human IgG for more than 15 months. Chromosome analysis showed the characteristics of human-mouse hybridoma. The cells of HMD4 were injected into the abdominal cavities of nude mice and 2-3 weeks later large quantities of ascites were obtained. Human IgG of lambda light chain was detected and purified from the ascites. The specificity of HMD4 human McAb was tested by ABC or PAP immunoperoxidase stainings of paraffin-embedded tissue sections, cryostat sections, cell smears of various tissues and different cancer cell lines. 60.5% (26/43) of epithelial ovarian cancers was positive, while nonepithelial ovarian cancers, most cancers from other organs and almost all nonmalignant and normal tissues were negative. The molecular weight of the antigen recognized by HMD4 was 55 KDa determined by Western blotting. The problems of maintaining the IgG secreting function of human-mouse hybridoma and its screening were also discussed.
将卵巢癌患者区域淋巴结中的淋巴细胞与小鼠骨髓瘤的非分泌细胞系（Sp2/0-Ag14）融合，使其永生化。融合率为0 - 87.5%。通过早期克隆和再克隆，建立了一个名为HMD4的杂交细胞系。它已稳定分泌人IgG超过15个月。染色体分析显示了人 - 鼠杂交瘤的特征。将HMD4细胞注入裸鼠腹腔，2 - 3周后获得大量腹水。从腹水中检测并纯化出λ轻链的人IgG。通过对石蜡包埋组织切片、冰冻切片、各种组织的细胞涂片以及不同癌细胞系进行ABC或PAP免疫过氧化物酶染色，检测HMD4人单克隆抗体的特异性。60.5%（26/43）的上皮性卵巢癌呈阳性，而非上皮性卵巢癌、大多数其他器官的癌症以及几乎所有非恶性和正常组织均为阴性。通过蛋白质印迹法确定HMD4识别的抗原分子量为55 kDa。还讨论了维持人 - 鼠杂交瘤IgG分泌功能及其筛选的问题。
