Brindley P J, Gam A A, Pearce E J, Poindexter R W, Neva F A
Laboratory of Parasitic Diseases, National Institute of Allergy and Infectious Diseases, Bethesda, MD 20892.
Mol Biochem Parasitol. 1988 Apr;28(3):171-80. doi: 10.1016/0166-6851(88)90001-1.
The surface and excretory/secretory (ES) antigens of the infective, filariform larva (L3) of Strongyloides stercoralis were identified. These studies provide a basis for the purification of these proteins as diagnostic allergens for human strongyloidiasis. The Mr values of the surface and ES molecules were determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and autoradiography, fluorography, or silver staining following the recovery of these molecules after the radiolabelling of living parasites. At least 10 surface proteins were radioiodinated extrinsically using chloroglycoluril as the catalyst for iodination, and then extracted with detergents and/or beta-mercaptoethanol. Several surface molecules of the L3 were immunogenic in humans, as determined by immunoprecipitation with sera (IHS) from infected patients. About 30 proteins were present in the ES preparation. Many ES antigens were labelled biosynthetically during the culture of larvae in media supplemented with either [35S]methionine or [14C]glucose. Furthermore, several of the surface proteins of the L3 were found with the ES antigens recoverable by culturing larvae in vitro. About 10 of the ES proteins were immunogenic as determined by immunoaffinity chromatography using IHS; and two of these antigens with Mr 50,000 and 90,000 incorporated [35S]methionine during culture of larvae. Moreover, some ES proteins were allergenic when tested in an in vitro assay of histamine release from basophils from infected humans or monkeys. The isotype of the homocytophilic antibodies involved in this immediate hypersensitivity assay, which is the basis of a diagnostic skin test for human strongyloidiasis, appears to be IgE.
已鉴定出粪类圆线虫感染性丝状蚴（L3）的表面抗原及排泄/分泌（ES）抗原。这些研究为纯化这些蛋白质作为人类粪类圆线虫病的诊断变应原奠定了基础。在对活寄生虫进行放射性标记后回收这些分子，然后通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳及放射自显影、荧光显影或银染法测定表面分子和ES分子的Mr值。至少10种表面蛋白通过使用氯甘脲作为碘化催化剂进行体外放射性碘化，然后用去污剂和/或β-巯基乙醇提取。用感染患者的血清进行免疫沉淀（IHS）测定，结果表明L3的几种表面分子在人体内具有免疫原性。ES制剂中存在约30种蛋白质。在添加了[35S]甲硫氨酸或[14C]葡萄糖的培养基中培养幼虫时，许多ES抗原被生物合成标记。此外，在体外培养幼虫时，发现L3的几种表面蛋白与可回收的ES抗原在一起。用IHS通过免疫亲和层析测定，约10种ES蛋白具有免疫原性；其中两种Mr为50,000和90,000的抗原在幼虫培养期间掺入了[35S]甲硫氨酸。此外，在对感染人类或猴子的嗜碱性粒细胞进行组胺释放的体外试验中，一些ES蛋白具有变应原性。参与这种速发型超敏反应试验（这是人类粪类圆线虫病诊断皮肤试验的基础）的亲同种细胞抗体的同种型似乎是IgE。
