抗阴道毛滴虫单克隆抗体的制备及其特性

Preparation and Characterization of Monoclonal Antibodies to T. Vaginalis

用杂交瘤技术建立了6株能稳定分泌抗阴道毛滴虫单克隆抗体的细胞株,选取其中3株进行了分析。3株单抗的效价分别是:ⅣA12B1和ⅣA22B1为250000ⅡB2A1为1250000。ⅣA12B1和ⅡB2A为IgG3抗体,ⅣA2281为IgGl抗体。位点分析及免疫酶染色发现,ⅣA12B1和ⅣA22B1抗同一抗原决定簇,所针对的抗原主要分布于膜上;ⅡB2A1针对的抗原主要分布于胞浆中。3株单抗针对的抗原均为糖蛋白。特异性试验发现,这3株单抗与人白细胞、阴道上皮细胞、白色念珠菌、大肠杆菌及人毛滴虫均无交叉反应。用制备的单克隆抗体,建立了试断滴虫性阴道炎的胶乳凝集试验(LAT)。研究表明,LAT是一种敏感、特异、简单和快速的方法。

Six hybrid cell lines producing monoclonal antibodies to T. vaginalis were established and three of them were characterized. The ascites produced by the three hybrid cell lines (Ⅱ B2A1, ⅣA12B1 and ⅣA22Bl) contained high-titred antibodies to T. vaginalis with one producing IgG1 and two producing IgG3. The cross-inhibition between the three McAbs revealed that both of ⅣA12B1 and ⅣA22Bl shared an antigenic determinant which was found to be present on membrane/flagella and I B2A1 recognized another antigenic determinant which existed in cytoplasm/f lagella by immunocytochem-istry of intact organisms. The antigens recognized by the three McAbs were found to be Sensitive to periodate oxidation. None of the three McAbs was cross-reactive to human white blood cells, vaginal epithelial cells, Hela cells, T. hominis, E. coli and C. albicans. The latex agglutination test established with McAb IVA22B1 showed a good correspondence with microscopic examination in more than 147 patients. The results reveal that preparation of the three McAbs may provide a potentially useful tool for diagnosis of Trichomonal vaginitis.