Bowman D D, Mika-Grieve M, Grieve R B
Department of Pathobiological Sciences, School of Veterinary Medicine, University of Wisconsin, Madison 53706.
Exp Parasitol. 1987 Dec;64(3):458-65. doi: 10.1016/0014-4894(87)90060-9.
When maintained in culture, the infective-stage larvae of Toxocara canis produce a group of excretory-secretory antigens. Monoclonal antibodies to these antigens have been produced and partially characterized. Hybridomas were made using spleens from mice that had been given 250 embryonated eggs of T. canis followed by immunization with excretory-secretory antigens. Monoclonal antibodies were first screened against excretory-secretory antigens using an indirect enzyme-linked immunosorbent assay. Those antibodies positive in this assay were then screened against the surfaces of formalin-fixed, infective-stage larvae using an indirect fluorescent antibody assay. The two monoclonal antibodies showing fluorescence were also tested against the surfaces of infective-stage larvae of Toxocara cati, Baylisascaris procyonis, Toxascaris leonina, Ascaris suum, a Porrocaecum sp., and Dirofilaria immitis. One of these two antibodies bound to the surface of T. canis and T. cati while the other bound only to the surface of T. canis; neither were reactive with the other ascaridoid larvae or the larvae of D. immitis. Enzyme-linked immunoelectrotransfer blotting techniques were used to demonstrate that the cross-reactive antibody recognized antigens with molecular weights of about 200 kDa while the more specific monoclonal antibody recognized antigens with approximate molecular weights of 80 kDa. The specificity of these two antibodies for T. canis and T. cati should prove helpful in the development of more specific assays for the diagnosis of visceral and ocular larva migrans.
犬弓首蛔虫感染期幼虫在体外培养时会产生一组排泄-分泌抗原。针对这些抗原的单克隆抗体已被制备并进行了部分特性分析。使用接种过250枚犬弓首蛔虫感染性虫卵的小鼠脾脏，再用排泄-分泌抗原进行免疫后制备杂交瘤。首先采用间接酶联免疫吸附测定法针对排泄-分泌抗原筛选单克隆抗体。然后使用间接荧光抗体测定法针对福尔马林固定的感染期幼虫表面对在该测定中呈阳性的抗体进行筛选。对显示荧光的两种单克隆抗体也针对猫弓首蛔虫、浣熊贝蛔虫、狮弓蛔虫、猪蛔虫、泡尾属某种线虫以及犬恶丝虫的感染期幼虫表面进行了检测。这两种抗体中的一种与犬弓首蛔虫和猫弓首蛔虫的表面结合，而另一种仅与犬弓首蛔虫的表面结合；二者均不与其他类圆线虫幼虫或犬恶丝虫幼虫发生反应。采用酶联免疫电转移印迹技术证明，交叉反应性抗体识别分子量约为200 kDa的抗原，而更具特异性的单克隆抗体识别分子量约为80 kDa的抗原。这两种抗体对犬弓首蛔虫和猫弓首蛔虫的特异性在开发更具特异性的内脏幼虫移行症和眼幼虫移行症诊断检测方法方面应会有所帮助。
