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针对克氏锥虫上鞭毛体鞭毛部分的单克隆抗体:补体介导的对锥鞭毛体的溶细胞活性及对小鼠的被动免疫保护作用

Monoclonal antibodies against the flagellar fraction of epimastigotes of Trypanosoma cruzi: complement-mediated lytic activity against trypomastigotes and passive immunoprotection in mice.

作者信息

Segura E L, Bua J, Rosenstein de Campanini A, Subias E, Esteva M, Moreno M, Ruiz A M

出版信息

Immunol Lett. 1986 Oct;13(4):165-71. doi: 10.1016/0165-2478(86)90050-7.

Abstract

Trypanosoma cruzi, the causative agent of Chagas' disease, is widely spread in Central and South America. The present report describes three monoclonal antibodies (mAbs) directed against the flagellar fraction of epimastigotes (Ffe) of the parasite, Tulahuén strain. The three mAbs were of IgG1 isotype. Indirect immunofluorescence assays revealed that the three mAbs bind to epimastigotes, the FCH-F8-1 and -3 bind to blood trypomastigotes (BT) and FCH-F8-1 is the only one that binds to amastigotes. Three different proteins of the parasite were recognized by the mAbs in immunoprecipitation assays: an 85 kDa of BT with the FCH-F8-1 mAb, a 40 kDa of Ffe with the FCH-F8-2 mAb, and a 90 kDa of Ffe and of BT with the FCH-F8-3 mAb. Positive complement mediated lysis (CML) of BT and metacyclic forms, obtained from the insect vector feces, were shown by the FCH-F8-1, while FCH-F8-3 only showed CML against the metacyclic trypomastigotes. FCH-F8-2 did not mediate any CML activity. Passive transference of the mAbs to BALB/c mice conferred protection, in terms of survival, ranging from 50 (FCH-F8-2 and -3) to 80% (FCH-F8-1) against a challenge with 1 X 10(3) BT. These results suggest that FCH-F8-1 may be a useful tool to purify proteins in order to investigate their role in immunoprotection experiments.

摘要

克氏锥虫是恰加斯病的病原体,广泛分布于中美洲和南美洲。本报告描述了三种针对该寄生虫图拉韦恩株无鞭毛体鞭毛部分(Ffe)的单克隆抗体(mAb)。这三种单克隆抗体均为IgG1亚型。间接免疫荧光分析表明,这三种单克隆抗体与无鞭毛体结合,FCH-F8-1和-3与血液锥鞭毛体(BT)结合,而FCH-F8-1是唯一与无鞭毛体结合的抗体。在免疫沉淀试验中,单克隆抗体识别出该寄生虫的三种不同蛋白质:FCH-F8-1单克隆抗体识别出一种85 kDa的血液锥鞭毛体蛋白,FCH-F8-2单克隆抗体识别出一种40 kDa的无鞭毛体鞭毛部分蛋白,FCH-F8-3单克隆抗体识别出一种90 kDa的无鞭毛体鞭毛部分蛋白和血液锥鞭毛体蛋白。FCH-F8-1显示出对血液锥鞭毛体和从昆虫媒介粪便中获得的循环后期形式的阳性补体介导裂解(CML),而FCH-F8-3仅对循环后期锥鞭毛体显示出CML。FCH-F8-2未介导任何CML活性。将单克隆抗体被动转移到BALB/c小鼠体内,在存活方面提供了保护,针对1×10³个血液锥鞭毛体的攻击,保护率从50%(FCH-F8-2和-3)到80%(FCH-F8-1)不等。这些结果表明,FCH-F8-1可能是纯化蛋白质以研究其在免疫保护实验中作用的有用工具。

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