Nickell S P, Keane M, So M
Department of Immunology and Infectious Diseases, Johns Hopkins School of Hygiene and Public Health, Baltimore, Maryland 21205.
Infect Immun. 1993 Aug;61(8):3250-8. doi: 10.1128/iai.61.8.3250-3258.1993.
We previously reported the isolation from immune mice of a panel of murine clonal T-cell lines which specifically recognize antigens expressed by the trypomastigote stage of the protozoan parasite Trypanosoma cruzi, the causative agent of human Chagas' disease. Our analysis indicated that distinct clones which recognize common as well as strain-specific antigenic determinants were represented. The immunoprotective potential of several of these T-cell clones was demonstrated by adoptive transfer of protection to naive syngeneic recipients. Here we report that these T-cell clones are all of the TH1 phenotype, as determined from their lymphokine secretion patterns. Significant levels of stimulatory activity for each clone were detected in trypomastigote supernatants, and the release of this activity was time and temperature dependent. Seven of 10 T-cell clones tested responded to nitrocellulose-immunoblotted trypomastigote proteins in the range of 90 to 47 kDa; no fewer than six distinct epitopes residing on at least five distinct polypeptide species were recognized by this panel of clones. Two clones (2G8 and 4B10) previously shown to protect in vivo responded to immunoblotted proteins in the range of 65 to 53 and 90 to 80 kD, respectively. Stimulatory activity for the latter clone was shown to be expressed on the surface of trypomastigotes and to bind specifically to wheat germ agglutinin, indicating that its target antigen is an 85-kDa trypomastigote surface glycoprotein.
我们先前报道过,从免疫小鼠中分离出一组鼠源克隆T细胞系,它们能特异性识别原生动物寄生虫克氏锥虫(人类恰加斯病的病原体)无鞭毛体阶段所表达的抗原。我们的分析表明,这些克隆代表了识别共同抗原决定簇以及菌株特异性抗原决定簇的不同克隆。通过将保护作用过继转移给同基因的未免疫受体,证实了其中几个T细胞克隆具有免疫保护潜力。在此我们报道,根据其淋巴因子分泌模式确定,这些T细胞克隆均为TH1表型。在无鞭毛体的培养上清液中检测到每个克隆都有显著水平的刺激活性,且这种活性的释放与时间和温度有关。在接受检测的10个T细胞克隆中,有7个对硝酸纤维素免疫印迹法检测的90至47 kDa范围内的无鞭毛体蛋白有反应;这组克隆识别出至少5种不同多肽上存在不少于6个不同的表位。先前显示在体内具有保护作用的两个克隆(2G8和4B10),分别对65至53 kDa和90至80 kDa范围内的免疫印迹蛋白有反应。已证明后一个克隆的刺激活性在无鞭毛体表面表达,并能与麦胚凝集素特异性结合,这表明其靶抗原是一种85 kDa的无鞭毛体表面糖蛋白。