【摘要】 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)的保护性免疫力和免疫保护性机制。方法　用重组纯化的Calpain抗原免疫小鼠 ,制备抗Calpain血清 ,将机械转化的日本血吸虫童虫和同种鼠激活的嗜酸性粒细胞共同培养 ,观察细胞对血吸虫童虫的黏附及对童虫的杀伤效果。结果　重组Calpain抗原免疫小鼠后 ,产生了极高的抗Calpain特异性抗体 ,并介导了同种鼠的嗜酸性粒细胞对日本血吸虫童虫的黏附。在 3 7°C ,5 %CO2 培养 48h后 ,3 3 .8%的日本血吸虫童虫被杀死 ,与对照组 14 .5 %的自然死亡率相比显著增高 (P <0 .0 1)。结论　Calpain特异性抗体介导了对日本血吸虫童虫的细胞毒作用 ,提示Calpain可能是日本血吸虫的一个疫苗候选分子 ,该分子的重组抗原有可能应用于血吸虫病诊断更多还原

【Abstract】 Objective To explore the efficacy and protective mechanism of calpain, a calcium activated neutral proteinase as a vaccine candidate molecule. Methods Anti calpain sera were prepared by immunized BALB/c mice with recombinant calpain. Anti calpain sera, schistosomula of Schistosoma japonicum , and mouse eosinophils were incubated at 37 °C , 5% CO 2 for 48 hours, and the adherence of eosinophils to schistosomula and its schistosomulacidal efficacy were observed. Results Mice immunized with recombinant calpain produced a high level IgG antibodies specific to the antigen immunized. Immunoblotting analysis showed murine anti recombinant calpain sera bound specifically to recombinant calpain. Ninety six percent of schistosomula were surrounded by cells when incubated with mouse eosinophils, and a significant number of dead schistosomula was observed at 48 hours when incubated with mouse serum and eosinophils as compared with control serum and eosinophils ( P <0.01). Conclusions Calpain antibodies induce antibody dependent cell adherence and killing effect of the schistosomula, suggesting that calpain of Schistosoma japonicum can not only be a vaccine candidate, but also a target antigen for immunodiagnosis of human schistosomiasis.更多还原