Stephens R S, Tam M R, Kuo C C, Nowinski R C
J Immunol. 1982 Mar;128(3):1083-9.
Nineteen independent hybrid cell lines that produce monoclonal antibodies to Chlamydia trachomatis surface antigens were prepared by the fusion of mouse myeloma cells with lymphocytes of mice that were immunized with C. trachomatis immunotypes B, C, and L2. Seven serologically distinct reaction patterns were detected by microimmunofluorescence (micro-IF) of elementary body (EB) preparations when culture fluids were tested against a panel of 18 chlamydial serotyping reference strains. These reaction patterns demonstrated genus-, species-, subspecies-, and type-specific distributions. Additionally, these antibodies were tested in parallel against reticulate body (RB) preparations of several chlamydial strains. Monoclonal antibodies that reacted with genus-specific antigens reacted preferentially with RB, whereas antibodies that reacted to species-, subspecies-, or type-specific antigens reacted equivalently to both RB and EB. Physiochemical characterization of antigens recognized by the different monoclonal antibodies was assessed by heat treatment, pronase digestion, periodate oxidation, and immuno-blot techniques. The genus-specific antigen was a heat-stable, pronase-resistant, and relatively periodate-sensitive component of less than 10,000 m.w. The species-, subspecies-, and type-specific antigens were heat stable, pronase sensitive, and periodate resistant. The antibodies that detected species- and subspecies-specific antigens predominantly reacted in immuno-blots with the 40,000 m.w. major outer membrane protein. These monoclonal antibodies now provide a new approach for the precise serologic classification and detection of different C. trachomatis strains.
通过将小鼠骨髓瘤细胞与用沙眼衣原体免疫型B、C和L2免疫的小鼠淋巴细胞融合,制备了19个产生抗沙眼衣原体表面抗原单克隆抗体的独立杂交细胞系。当用培养液检测一组18种衣原体血清型参考菌株时,通过对原体(EB)制剂进行微量免疫荧光(微量IF)检测,发现了7种血清学上不同的反应模式。这些反应模式显示出属、种、亚种和型特异性分布。此外,还将这些抗体与几种衣原体菌株的网状体(RB)制剂进行了平行检测。与属特异性抗原反应的单克隆抗体优先与RB反应,而与种、亚种或型特异性抗原反应的抗体对RB和EB的反应相同。通过热处理、链霉蛋白酶消化、高碘酸盐氧化和免疫印迹技术评估了不同单克隆抗体识别的抗原的理化特性。属特异性抗原是一种热稳定、抗链霉蛋白酶且相对对高碘酸盐敏感的成分,分子量小于10,000。种、亚种和型特异性抗原热稳定、对链霉蛋白酶敏感且对高碘酸盐有抗性。检测种和亚种特异性抗原的抗体在免疫印迹中主要与40,000分子量的主要外膜蛋白反应。这些单克隆抗体现在为精确的血清学分类和检测不同的沙眼衣原体菌株提供了一种新方法。