Jaffe C L, Rachamim N
Department of Biophysics, MacArthur Center for Molecular Biology of Tropical Diseases, Weizmann Institute of Science, Rehovot, Israel.
Infect Immun. 1989 Dec;57(12):3770-7. doi: 10.1128/iai.57.12.3770-3777.1989.
Monoclonal antibodies were produced against gamma-irradiated amastigotes of Leishmania major. Five antibodies (T16 through T20) were selected which reacted in enzyme-linked immunoassays with the intracellular stage of the parasite. These antibodies did not react with promastigotes of L. major or Leishmania donovani. One of the monoclonal antibodies (T16) reacted with amastigotes of Leishmania mexicana amazonensis and L. donovani. Western blotting (immunoblotting) and immunoprecipitation of [35S]methionine-labeled amastigotes demonstrates that T16 reacted with multiple L. major amastigote components between 12 and 180 kilodaltons. Antibody T20 was shown to recognize a low-molecular-mass doublet (less than 26 kilodaltons) in both [14C]leucine- and [35S]methionine-labeled amastigotes. A protein of less than 180 kilodaltons was also weakly recognized by T17, T19, and T20 in metabolically labeled amastigotes. This protein reacted strongly with T16. The reactive antigens could be identified on the surface of amastigotes isolated from the lesions of infected mice and on newly transformed amastigotes within 24 h after the infection of mouse peritoneal macrophages by promastigotes. These monoclonal antibodies should prove useful for the diagnosis of L. major in human tissue biopsies.
制备了针对经γ射线照射的硕大利什曼原虫无鞭毛体的单克隆抗体。筛选出了五种抗体（T16至T20），它们在酶联免疫测定中与该寄生虫的细胞内阶段发生反应。这些抗体不与硕大利什曼原虫或杜氏利什曼原虫的前鞭毛体发生反应。其中一种单克隆抗体（T16）与亚马逊利什曼原虫和杜氏利什曼原虫的无鞭毛体发生反应。对[35S]甲硫氨酸标记的无鞭毛体进行蛋白质印迹法（免疫印迹）和免疫沉淀表明，T16与分子量在12至180千道尔顿之间的多种硕大利什曼原虫无鞭毛体成分发生反应。抗体T20在[14C]亮氨酸和[35S]甲硫氨酸标记的无鞭毛体中均能识别一种低分子量双峰（小于26千道尔顿）。在代谢标记的无鞭毛体中，T17, T19和T20也能微弱识别一种小于180千道尔顿的蛋白质。该蛋白质与T16反应强烈。在从感染小鼠病变部位分离出的无鞭毛体表面以及前鞭毛体感染小鼠腹腔巨噬细胞后24小时内新转化的无鞭毛体表面，均可鉴定出反应性抗原。这些单克隆抗体应有助于在人体组织活检中诊断硕大利什曼原虫感染。
