James S L, Pearce E J, Sher A
J Immunol. 1985 May;134(5):3432-8.
A single intradermal injection of frozen and thawed schistosomula in conjunction with the bacterial adjuvant Mycobacterium bovis strain Bacille Calmette Guerin, Phipps substrain (BCG) induced significant levels of resistance to challenge Schistosoma mansoni infection in C57BL/6 mice. Immunization with the aqueous fraction remaining after 100,000 X G centrifugation of the larval lysate was also protective under these conditions, suggesting that some immunogenic determinants may not be membrane associated. Frozen-thawed cercariae and soluble components of adult worms also protected against challenge infection in these experiments. These observations indicate that soluble immunogens are present in both early and late developmental stages of the parasite, and therefore may be good candidate antigens for an immunochemically defined vaccine against schistosomiasis. Induction of humoral reactivity against soluble or membrane antigens was examined in mice protected against cercarial challenge by prior exposure to frozen-thawed larvae, soluble larval, or soluble adult antigens plus BCG. Animals that were immunized with frozen-thawed larvae produced low but significant levels of antibodies against larval surface antigens when examined by indirect immunofluorescence or by immunoprecipitation of surface-labeled schistosomula. Mice immunized with soluble antigens, however, showed negligible antibody reactivity against surface membrane antigens. Because mice immunized with soluble antigens were resistant to challenge infection, these results strongly suggest that anti-surface membrane reactivity is not required in the mechanism of protective immunity in this model. Sera from mice immunized with either total freeze-thaw larval lysate or soluble schistosome extracts all showed strong reactivity against soluble antigens, as detected by ELISA. Western blot analysis showed these antisera to react with a restricted number of high m.w. antigens that were present both in schistosomula and in adult worms. These antigens are therefore likely to play a major role in the development of resistance in this model as immunogens and/or as targets of protective immune response.
将冷冻解冻的血吸虫童虫与细菌佐剂牛分枝杆菌卡介苗(Phipps亚株,BCG)进行单次皮内注射,可诱导C57BL/6小鼠对曼氏血吸虫攻击感染产生显著水平的抗性。在这些条件下,用幼虫裂解物经100,000×G离心后剩余的水相部分进行免疫也具有保护作用,这表明一些免疫原性决定簇可能与膜无关。在这些实验中,冷冻解冻的尾蚴和成虫的可溶性成分也能预防攻击感染。这些观察结果表明,可溶性免疫原存在于寄生虫的早期和晚期发育阶段,因此可能是针对血吸虫病的免疫化学定义疫苗的良好候选抗原。在预先暴露于冷冻解冻的幼虫、可溶性幼虫或可溶性成虫抗原加BCG而免受尾蚴攻击的小鼠中,检测了针对可溶性或膜抗原的体液反应性的诱导情况。当通过间接免疫荧光或表面标记的血吸虫童虫的免疫沉淀检测时,用冷冻解冻的幼虫免疫的动物产生了低但显著水平的针对幼虫表面抗原的抗体。然而,用可溶性抗原免疫的小鼠对表面膜抗原的抗体反应可忽略不计。由于用可溶性抗原免疫的小鼠对攻击感染具有抗性,这些结果强烈表明在该模型的保护性免疫机制中不需要抗表面膜反应性。用总冷冻解冻幼虫裂解物或可溶性血吸虫提取物免疫的小鼠血清,通过ELISA检测均显示出对可溶性抗原的强反应性。蛋白质印迹分析表明,这些抗血清与血吸虫童虫和成虫中都存在的数量有限的高分子量抗原发生反应。因此,这些抗原作为免疫原和/或保护性免疫反应的靶标,可能在该模型的抗性发展中起主要作用。