Northern C, Grove D I, Warton A, Lovegrove F T
Department of Medicine, University of Western Australia, Nedlands.
Clin Exp Immunol. 1989 Mar;75(3):487-92.
Various methods of radioiodination were compared for their efficacy in labelling the surface of Strongyloides ratti infective larvae and adult worms. The Iodogen method was chosen as the optimal technique for this parasite. The surface location of 125-iodine was confirmed with light microscope autoradiography of transverse sections of labelled worms. Stage-specific surface components were identified when the sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) profiles of infective larvae and adult worms were compared. Labelled surface molecules were solubilized with either the non-ionic detergent Triton-X-100, the anionic detergents sodium deoxycholate (DOC) or sodium dodecyl sulphate (SDS), or the cationic detergent cetyl trimethyl-ammonium bromide (CTAB). The CTAB extract yielded most labelled proteins that retained their antigenicity in an immunoprecipitation assay with hyperimmune mouse sera. Immunoprecipitation analysis with stage-specific mouse sera revealed that the surface of infective larvae is immunogenic and that there are no cross-reactions with adult worms. Adult worms resident in the intestine were not found to be immunogenic and showed a complete absence of reactivity. Antigenic determinants shared between S. ratti and S. stercoralis were identified. Patients infected with S. stercoralis precipitated bands with molecular weights 32 and 34 kD which were not reactive with normal sera. These reactions suggest the potential usefulness of the surface of S. ratti as a source of diagnostic antigens.
比较了各种放射性碘化方法对鼠类圆线虫感染性幼虫和成虫表面进行标记的效果。碘珠法被选为针对这种寄生虫的最佳技术。通过对标记虫体横切面的光学显微镜放射自显影,证实了¹²⁵碘的表面定位。比较感染性幼虫和成虫的十二烷基硫酸钠聚丙烯酰胺凝胶电泳(SDS-PAGE)图谱时,鉴定出了阶段特异性表面成分。用非离子去污剂Triton-X-100、阴离子去污剂脱氧胆酸钠(DOC)或十二烷基硫酸钠(SDS),或阳离子去污剂十六烷基三甲基溴化铵(CTAB)溶解标记的表面分子。CTAB提取物产生了大多数标记蛋白,这些蛋白在与超免疫小鼠血清的免疫沉淀试验中保留了其抗原性。用阶段特异性小鼠血清进行的免疫沉淀分析表明,感染性幼虫的表面具有免疫原性,并且与成虫没有交叉反应。未发现寄生于肠道的成虫具有免疫原性,且完全没有反应性。鉴定出了鼠类圆线虫和粪类圆线虫之间共有的抗原决定簇。感染粪类圆线虫的患者沉淀出分子量为32和34kD的条带,这些条带与正常血清无反应。这些反应表明鼠类圆线虫表面作为诊断抗原来源具有潜在用途。