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小鼠单克隆抗体的制备及宿主对曼氏血吸虫尾蚴髋臼前腺蛋白酶免疫反应的证据

Preparation of mouse monoclonal antibodies and evidence for a host immune response to the preacetabular gland proteinase of Schistosoma mansoni cercariae.

作者信息

Pino-Heiss S, Petitt M, Beckstead J H, McKerrow J H

出版信息

Am J Trop Med Hyg. 1986 May;35(3):536-43. doi: 10.4269/ajtmh.1986.35.536.

Abstract

Forty-six monoclonal antibodies were produced against the preacetabular gland secretions of Schistosoma mansoni cercariae by two different immunization protocols. These antibodies were of both the IgG and the IgM classes. One IgM monoclonal antibody (Ia4D6) was further characterized. It was specific to the cercarial stage by ELISA and showed specific binding to the 30,000 Mr proteinase in crude cercarial secretions by Western blot analysis. Preincubation of this antibody with purified cercarial proteinase resulted in inhibition of proteolytic activity, and it mediated complement-dependent cytotoxicity to cercariae in vitro. Immunoperoxidase staining of cercariae localized this antibody to vesicles visible within the preacetabular glands and their secretory ducts, and to secreted material. ELISA and Western blot analysis also showed that sera from infected mice and patients with schistosomiasis reacted with the cercarial proteinase. These studies demonstrate that a proteinase secreted into the host by invading cercariae is immunogenic and provide a monoclonal antibody probe for further characterization of its structure and function.

摘要

通过两种不同的免疫方案,制备了46种针对曼氏血吸虫尾蚴前吸盘腺分泌物的单克隆抗体。这些抗体属于IgG和IgM两类。对一种IgM单克隆抗体(Ia4D6)进行了进一步鉴定。通过ELISA法,它对尾蚴阶段具有特异性,并且通过蛋白质印迹分析显示其与粗制尾蚴分泌物中的30,000 Mr蛋白酶具有特异性结合。该抗体与纯化的尾蚴蛋白酶预孵育会导致蛋白水解活性受到抑制,并且它在体外介导对尾蚴的补体依赖性细胞毒性。尾蚴的免疫过氧化物酶染色将该抗体定位到前吸盘腺及其分泌导管内可见的小泡以及分泌物质上。ELISA和蛋白质印迹分析还表明,感染小鼠和血吸虫病患者的血清与尾蚴蛋白酶发生反应。这些研究表明,入侵的尾蚴分泌到宿主体内的一种蛋白酶具有免疫原性,并为进一步表征其结构和功能提供了一种单克隆抗体探针。

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