Hillyer G V, Pacheco E
Am J Trop Med Hyg. 1986 Jul;35(4):777-85. doi: 10.4269/ajtmh.1986.35.777.
Schistosoma haematobium soluble egg antigen (ShSEA) was prepared from eggs isolated from the livers of hamsters or mice infected for at least 3 months. Immunoaffinity purified S. haematobium egg antigens (ShSh) were isolated by first passing ShSEA through a column containing anti-S. mansoni hamster IgG coupled to CNBr-activated Sepharose 4B, and recycling the unbound fraction until no more bound material could be eluted with an acid wash. The unbound fraction was then filtered through a second antibody affinity column containing anti-S. haematobium hamster IgG, and in the acid eluate the ShSh antigens were obtained. This antigenic preparation was shown by PAGE to contain at least 6 distinct bands ranging in molecular weight (Mr) from 116 to less than 31 Kd. A 40 Kd polypeptide was identified by both silver staining and EITB as specific for S. haematobium eggs. In addition, a 55 Kd worm-egg shared antigen was identified as a prominent band in EITB expressed during a primary S. haematobium hamster infection. The sera from hamsters harboring patent S. haematobium or S. mansoni infections were reacted by ELISA with ShSh antigens. The anti-Sh sera showed significantly higher absorbance values than the anti-Sm sera, demonstrating that only a minor population of S. mansoni cross-reactive egg antigens is still present in the ShSh antigens. Sera collected weekly for 13 weeks from hamsters with a primary infection of S. haematobium were then tested by ELISA against ShSh, ShSEA and SmSEA antigens. Antibody levels against both ShSEA and SmSEA were shown to increase early in infection (2 weeks). Moreover, antibody levels to ShSh did not increase until week 5 post-infection. These findings suggest that the purification procedure utilized results in the elimination of most of the S. mansoni worm antigens cross-reactive with S. haematobium eggs. The ShSh antigens had shown a high degree of sensitivity and stage-species specificity also suggesting their potential as antigens for the immunodiagnosis of schistosomiasis haematobia.
埃及血吸虫可溶性虫卵抗原(ShSEA)是从感染至少3个月的仓鼠或小鼠肝脏中分离的虫卵制备而来。免疫亲和纯化的埃及血吸虫虫卵抗原(ShSh)的分离方法是,首先将ShSEA通过一个含有与溴化氰活化的琼脂糖4B偶联的抗曼氏血吸虫仓鼠IgG的柱子,然后将未结合部分循环使用,直到用酸洗不再能洗脱结合物质。然后将未结合部分通过第二个含有抗埃及血吸虫仓鼠IgG的抗体亲和柱过滤,在酸洗脱液中获得ShSh抗原。通过聚丙烯酰胺凝胶电泳(PAGE)显示,这种抗原制剂包含至少6条不同的条带,分子量(Mr)范围从116到小于31千道尔顿。通过银染和免疫印迹法(EITB)鉴定出一条40千道尔顿的多肽对埃及血吸虫虫卵具有特异性。此外,在埃及血吸虫仓鼠初次感染期间表达的免疫印迹中,一条55千道尔顿的虫-卵共同抗原被鉴定为一条突出条带。携带显性埃及血吸虫或曼氏血吸虫感染的仓鼠血清通过酶联免疫吸附测定(ELISA)与ShSh抗原反应。抗Sh血清显示出比抗Sm血清显著更高的吸光度值,表明在ShSh抗原中仅存在少量与曼氏血吸虫交叉反应的虫卵抗原。然后,对初次感染埃及血吸虫的仓鼠每周收集13周的血清进行ELISA检测,以检测ShSh、ShSEA和SmSEA抗原。结果显示,针对ShSEA和SmSEA的抗体水平在感染早期(2周)就升高。此外,针对ShSh的抗体水平直到感染后第5周才升高。这些发现表明,所采用的纯化程序导致消除了大多数与埃及血吸虫虫卵交叉反应的曼氏血吸虫虫体抗原。ShSh抗原显示出高度的敏感性和阶段-种特异性,这也表明它们作为埃及血吸虫病免疫诊断抗原的潜力。