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盘尾丝虫一种免疫显性糖蛋白抗原的特性及其在其他丝虫线虫和秀丽隐杆线虫中的同源物

Characterisation of an immunodominant glycoprotein antigen of Onchocerca volvulus with homologues in other filarial nematodes and Caenorhabditis elegans.

作者信息

Tree T I, Gillespie A J, Shepley K J, Blaxter M L, Tuan R S, Bradley J E

机构信息

Department of Biology, Imperial College, London, UK.

出版信息

Mol Biochem Parasitol. 1995 Feb;69(2):185-95. doi: 10.1016/0166-6851(94)00204-z.

Abstract

The full-length cDNA corresponding to an Onchocerca volvulus antigen, OvMBP/11, which had been selected as a serodiagnostic tool was isolated, sequenced, and the native antigen encoded by the cDNA characterised. The cDNA encodes a protein of 20.5 kDa (termed Ov 20) containing a putative signal peptide. Southern blot analysis indicates that there is only a single O. volvulus gene corresponding to Ov 20 but it has significant sequence similarity to genes corresponding to two 20.5-kDa predicted proteins of Caenorhabditis elegans. Homologues of the Ov 20 gene were detected at high stringency by Southern blot in the other Onchocerca species O. gibsoni, and O. gutturosa and at lower stringency in the related filarial nematodes Brugia malayi and Acanthocheilonema viteae. The Ov 20 native antigen has two molecular mass forms, 20 and 22 kDa, in all the life cycle stages studied. These isoforms have different levels of N-linked glycosylation on a peptide backbone of 17.5 kDa. Immunolocalisation and in situ hybridisation studies demonstrated that Ov 20 is transcribed and translated in the body wall of adult females and also in microfilariae, third and fourth stage larvae. Antigen was detected in the supernatant of in vitro cultured adult female nematodes. The B. malayi and A. viteae homologues are antigenically cross-reactive to Ov 20, share the same size peptide backbone but differ in their degree of glycosylation.

摘要

分离、测序了与盘尾丝虫抗原OvMBP/11对应的全长cDNA,该抗原已被选作血清诊断工具,并对cDNA编码的天然抗原进行了特性分析。该cDNA编码一种20.5 kDa的蛋白质(称为Ov 20),含有一个推定的信号肽。Southern印迹分析表明,只有一个与Ov 20对应的盘尾丝虫基因,但它与秀丽隐杆线虫两个预测的20.5 kDa蛋白质对应的基因具有显著的序列相似性。通过Southern印迹在其他盘尾丝虫物种吉氏盘尾丝虫和喉瘤盘尾丝虫中以高严谨度检测到Ov 20基因的同源物,在相关丝虫马来布鲁线虫和旋盘尾丝虫中以较低严谨度检测到。在所研究的所有生命周期阶段,Ov 20天然抗原具有两种分子量形式,即20 kDa和22 kDa。这些同工型在17.5 kDa的肽主链上具有不同水平的N-连接糖基化。免疫定位和原位杂交研究表明,Ov 20在成年雌性虫体壁以及微丝蚴、第三和第四期幼虫中进行转录和翻译。在体外培养的成年雌性线虫的上清液中检测到抗原。马来布鲁线虫和旋盘尾丝虫的同源物与Ov 20具有抗原交叉反应性,共享相同大小的肽主链,但糖基化程度不同。

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