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对具有潜在免疫诊断重要性的普通圆线虫表面抗原的特性分析。

Characterisation of surface antigens of Strongylus vulgaris of potential immunodiagnostic importance.

作者信息

Nichol C, Masterson W J

机构信息

Department of Clinical Veterinary Medicine, University of Cambridge, U.K.

出版信息

Mol Biochem Parasitol. 1987 Aug;25(1):29-38. doi: 10.1016/0166-6851(87)90015-6.

Abstract

When antigens prepared by detergent washes of Strongylus vulgaris and Parascaris equorum were probed in an enzyme-linked immunosorbent assay test with horse sera from single species infections of S. vulgaris and P. equorum, a high degree of cross-reaction between the species was demonstrated. Western blot analysis of four common horse nematode species showed a large number of common antigens when probed with horse infection sera. Antisera raised in rabbits against the four species, including S. vulgaris, were also found to cross-react considerably. Rabbit anti-S. vulgaris sera were affinity adsorbed over a series of affinity chromatography columns, bound with cross-reactive surface antigens, to obtain S. vulgaris-specific antisera and thereby identify S. vulgaris-specific antigens by Western blotting. These studies revealed potentially specific antigens of apparent molecular weights of 100,000, 52,000, and 36,000. Of these bands, only the 52 kDa and 36 kDa appeared to be found on the surface as judged by 125I-labelling of intact worms by the Iodogen method, although neither protein was immunoprecipitated by horse infection sera. Finally, immunoprecipitation of in vitro translated proteins derived from larval S. vulgaris RNA suggests that two proteins may be parasite-derived. These findings are discussed both with respect to the surface of S. vulgaris and to the use of these species-specific antigens in immunodiagnosis.

摘要

当用来自普通圆线虫(Strongylus vulgaris)和马副蛔虫(Parascaris equorum)单物种感染马的血清,在酶联免疫吸附测定试验中检测经去污剂洗涤制备的普通圆线虫和马副蛔虫抗原时,发现这两个物种之间存在高度交叉反应。对四种常见马线虫物种进行的蛋白质印迹分析表明,用马感染血清检测时存在大量共同抗原。还发现用包括普通圆线虫在内的这四种物种免疫家兔产生的抗血清也有相当程度的交叉反应。将兔抗普通圆线虫血清在一系列与交叉反应性表面抗原结合的亲和层析柱上进行亲和吸附,以获得普通圆线虫特异性抗血清,从而通过蛋白质印迹法鉴定普通圆线虫特异性抗原。这些研究揭示了表观分子量分别为100,000、52,000和36,000的潜在特异性抗原。在这些条带中,通过碘代甘氨酸法对完整虫体进行125I标记判断,只有52 kDa和36 kDa的条带似乎出现在表面,尽管这两种蛋白质均未被马感染血清免疫沉淀。最后,对源自普通圆线虫幼虫RNA的体外翻译蛋白质进行免疫沉淀表明,可能有两种蛋白质来自寄生虫。将结合普通圆线虫的表面以及这些物种特异性抗原在免疫诊断中的应用对这些发现进行了讨论。

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