Raleigh J M, Brandon M R, Meeusen E
Centre for Animal Biotechnology, University of Melbourne, Parkville, Victoria, Australia.
Parasite Immunol. 1996 Mar;18(3):125-32. doi: 10.1046/j.1365-3024.1996.d01-63.x.
Three monoclonal antibodies, Hc2, Hc22 and Hc6, were produced against a surface extract of L3 Haemonchus contortus and screened against both free living and parasitic stages of the parasite. Hc2 and Hc22 were of IgG2c isotype and their target epitopes were insensitive to periodate treatment. Hc6 was of IgM isotype and its reactivity was sensitive to periodate treatment of the antigen, suggesting that Hc6 is specific for a carbohydrate epitope. Western blotting of larval extracts and staining of live worms demonstrated that Hc2 was specific for the surface of second stage larvae and that the epitope was still present on the protective sheath of third stage larvae but absent from the L3 cuticle itself. Hc22 was found to be specific for a 70-90 kDa antigen on the surface of third stage infective larvae and cross reacted with a higher molecular weight species present on the surface of third stage Teladorsagia circumcincta. Hc6 showed strong binding to three high molecular weight proteins on immunoblots of third stage larvae only, but in contrast to Hc2 and Hc22, showed no binding to the surface of live larvae. These studies demonstrate unique expression patterns of stage specific antigens on the surface of free living and parasitic H. contortus larvae.
制备了三种针对捻转血矛线虫L3期表面提取物的单克隆抗体,即Hc2、Hc22和Hc6,并对该寄生虫的自由生活阶段和寄生阶段进行了筛选。Hc2和Hc22属于IgG2c同种型,其靶表位对高碘酸盐处理不敏感。Hc6属于IgM同种型,其反应性对抗原的高碘酸盐处理敏感,这表明Hc6对碳水化合物表位具有特异性。对幼虫提取物进行蛋白质印迹分析以及对活虫进行染色表明,Hc2对二期幼虫的表面具有特异性,并且该表位在三期幼虫的保护鞘上仍然存在,但在L3角质层本身不存在。发现Hc22对三期感染性幼虫表面的一种70 - 90 kDa抗原具有特异性,并与存在于三期环形泰勒虫表面的一种分子量更高的物质发生交叉反应。Hc6仅在三期幼虫的免疫印迹上显示与三种高分子量蛋白有强结合,但与Hc2和Hc22不同的是,它不与活幼虫的表面结合。这些研究证明了自由生活和寄生的捻转血矛线虫幼虫表面阶段特异性抗原的独特表达模式。