Cochrane A H, Santoro F, Nussenzweig V, Gwadz R W, Nussenzweig R S
Proc Natl Acad Sci U S A. 1982 Sep;79(18):5651-5. doi: 10.1073/pnas.79.18.5651.
Nine monoclonal antibodies against surface antigens of sporozoites of the simian malaria parasite Plasmodium knowlesi were produced by fusion of plasmacytoma cells with spleen cells of a mouse immunized with the parasites. Immunoprecipitation of extracts of [35S]methionine-labeled sporozoites with seven of the monoclonals identified the same three polypeptides with apparent molecular weights of 52,000 (Pk52), 50,000 (Pk50) and 42,000 (Pk42). These antigens also were recognized by serum of a rhesus monkey immunized with and protected against P. knowlesi sporozoites. Pulse--chase experiments indicated that the higher molecular weight proteins are precursors of Pk42. As shown by trypsin treatment of viable sporozoites, Pk42 is a surface antigen whereas Pk52 and Pk50 appear to be intracellular. Three of the monoclonal antibodies also reacted with a membrane antigen of sporozoites of another simian malaria, P. cynomolgi, and one monoclonal antibody reacted with sporozoites of human malaria, P. falciparum. When assayed for sporozoite neutralizing activity, most of the antibodies and their Fab fragments, which recognize Pk52, Pk50, and Pk42, abolished parasite infectivity.
通过将骨髓瘤细胞与用疟原虫免疫的小鼠脾细胞融合,制备了9种针对猴疟原虫诺氏疟原虫子孢子表面抗原的单克隆抗体。用其中7种单克隆抗体对[35S]甲硫氨酸标记的子孢子提取物进行免疫沉淀,鉴定出3种表观分子量分别为52,000(Pk52)、50,000(Pk50)和42,000(Pk42)的相同多肽。这些抗原也能被用诺氏疟原虫子孢子免疫并获得保护的恒河猴血清识别。脉冲追踪实验表明,分子量较高的蛋白质是Pk42的前体。如用胰蛋白酶处理活子孢子所示,Pk42是一种表面抗原,而Pk52和Pk50似乎是细胞内抗原。其中3种单克隆抗体也与另一种猴疟——食蟹猴疟原虫的子孢子膜抗原发生反应,1种单克隆抗体与人类疟疾疟原虫——恶性疟原虫的子孢子发生反应。在检测子孢子中和活性时,大多数识别Pk52、Pk50和Pk42的抗体及其Fab片段消除了寄生虫的感染性。