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巴西日圆线虫分泌型乙酰胆碱酯酶的免疫细胞化学定位

Immunocytochemical localization of secretory acetylcholinesterase of the parasitic nematode Nippostrongylus brasiliensis.

作者信息

Nakazawa M, Yamada M, Uchikawa R, Arizono N

机构信息

Department of Medical Zoology, Kyoto Prefectural University of Medicine, Japan.

出版信息

Cell Tissue Res. 1995 Apr;280(1):59-64. doi: 10.1007/BF00304511.

Abstract

Various parasitic nematodes secrete acetylcholinesterase (AChE). In this study, the localization of AChE in the nematode Nippostrongylus brasiliensis and the secretory forms of AChE in culture fluid were examined. A thiocholine method revealed that AChE activity was localized in the subventral glands, which have a secretory and excretory function via a duct connected to the excretory pore. By electron microscopy, AChE activity was found mainly in the matrix of secretory granules, and sometimes in the Golgi apparatus in the subventral gland cells. These results show that nematode AChE is produced and stored in the subventral glands. Monoclonal antibodies against AChE of human erythrocytes or electric rays also bound to the nematode subventral gland, suggesting immuno-cross-reactivity of AChE among these species. When AChE activity in the nematode excretory-secretory product was examined by SDS polyacrylamide gel electrophoresis combined with the thiocholine method, intense activity was demonstrated as a single band at 74 kDa. Immunoblot analysis showed specific recognition of this molecule by IgE and IgG1 antibodies, but not by IgG2a antibody, in nematode-infected rat sera. These results indicate that the nematode AChE molecule produced in and secreted from the subventral glands is antigenic for the production of IgE/IgG1 in host animals.

摘要

多种寄生线虫会分泌乙酰胆碱酯酶(AChE)。在本研究中,检测了巴西日圆线虫中AChE的定位以及培养液中AChE的分泌形式。硫代胆碱法显示,AChE活性定位于腹侧下腺,腹侧下腺通过与排泄孔相连的导管具有分泌和排泄功能。通过电子显微镜观察,AChE活性主要存在于分泌颗粒的基质中,有时也存在于腹侧下腺细胞的高尔基体中。这些结果表明,线虫AChE是在腹侧下腺中产生并储存的。针对人红细胞或电鳐的AChE的单克隆抗体也与线虫腹侧下腺结合,表明这些物种之间的AChE具有免疫交叉反应性。当通过SDS聚丙烯酰胺凝胶电泳结合硫代胆碱法检测线虫排泄-分泌产物中的AChE活性时,在74 kDa处有一条强活性条带。免疫印迹分析表明,在感染线虫的大鼠血清中,IgE和IgG1抗体可特异性识别该分子,而IgG2a抗体则不能。这些结果表明,在腹侧下腺中产生并分泌的线虫AChE分子对于宿主动物产生IgE/IgG1具有抗原性。

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