日本血吸虫疫苗候选分子Calpain的原位表达

Location and characteristic of Calpain,candidate molecule of Schistosoma japonicumvaccine

目的 探讨日本血吸虫疫苗候选分子———钙离子激活的中性蛋白激酶 (Calpain)在日本血吸虫体内的表达部位及其特征。方法 用重组纯化的Calpain抗原免疫大鼠 ,制备抗Calpain血清 ,从感染鼠灌流分离的日本血吸虫成虫被切片 ,用羊抗大鼠IgG抗体作为二抗进行免疫组织化学分析 ,观察Calpain蛋白在日本血吸虫成虫体内的表达部位。 结果 重组Calpain抗原免疫大鼠后 ,产生了一个极高的抗Calpain特异性抗体 ,并在日本血吸虫成虫切片上识别自然的日本血吸虫Calpain ,且大多数表达在成虫真皮层。结论 日本血吸虫钙离子激活的中性蛋白激酶 (Calpain)主要分布在成虫肌肉和真皮层 ,提示Cal pain可能是日本血吸虫的一个疫苗候选分子 ,并提示这个分子的重组抗原有可能应用于血吸虫病诊断。

Aim To detect the location of calpain ,a calcium-activated neutral proteinase as a vaccine candidate molecule in adult of Schistosoma japonicum.Methods Anti calpain sera were prepared by immunized rat with recombinant calpain adult worm sections of Schistosoma japonicum were analysed by immunohistochemistry with Anti calpain sera,the expression place of calpain in adult worms of Schistosoma japonicum were observed.Results Rat immunized by recombinant calpain produced a high level IgG antibodies specific to the immunized antigen.Immunoblotting analysis showed rat anti recombinant calpain sera binding specifically to recombinant calpain.The schistosome calpain were localized in the surface of syncytial epithelium and underlying musculature.Conclusion Most of the schistosome calpain localize in the surface of syncytial epithelium,the calpain of Schistosoma japonicum seems to be not only a vaccine candidate,but also a target as antigen for immunodiagnosis of human schistosomiasis