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目的 通过比较分析日本血吸虫不同发育阶段抗原的异同 ,并以致弱尾蚴免疫血清识别不同发育阶段抗原 ,筛选具有保护性免疫作用的抗原分子。方法 分离制备日本血吸虫未成熟虫卵、成熟虫卵、尾蚴、雄性成虫、雌性成虫等不同发育阶段的抗原 ;采用ELISA分别测定正常尾蚴感染兔血清、紫外线照射致弱尾蚴免疫兔血清抗原的抗体水平 ;通过SDS PAGE电泳和免疫印迹技术 ,分析正常尾蚴感染血清与紫外线照射致弱尾蚴免疫兔血清对不同发育阶段的免疫反应性。结果与结论 SDS-PAGE结果显示未成熟虫卵抗原、成熟虫卵抗原、尾蚴抗原、雄性成虫抗原、雌性成虫抗原蛋白之间互有异同。间接ELISA试验显示致弱尾蚴免疫血清同样能诱导宿主产生高滴度的抗体水平。致弱尾蚴免疫血清共筛选出 2 3种具免疫学活性的抗原分子 ,分子量分别为 :2 4 .5kDa、2 8kDa、36kDa、4 3kDa、4 4kDa、4 8kDa、5 4kDa、5 8.5kDa、6 0kDa、6 2kDa、6 6kDa、6 8kDa、70kDa、75kDa、79kDa、85kDa、87kDa、91kDa、95kDa、97kDa、10 5kDa、10 8kDa、110kDa。其中未成熟虫卵和成熟虫卵抗原各占 11种 ,尾蚴抗原分子 4种 ,雄性成虫抗原分子 3种 ,雌性成虫抗原分子 6种 ,提示这些抗原分子可能为抗日本血吸虫病疫苗候选分子的重要靶标
OBJECTIVE: To compare and analyze the similarities and differences of antigens in different developmental stages of Schistosoma japonicum, and to identify the antigens of different developmental stages by immunofluorescence with weak cercariae, to screen antigen molecules with protective immunity. Methods The antigens of different stages of immature eggs, mature eggs, cercariae, male adults and female adults of Schistosoma japonicum were isolated and purified. ELISA was used to detect the serum levels of antibodies against rabbits immunized with cercariae of cercariae by ELISA SDS-PAGE electrophoresis and Western blotting were used to analyze the immune reactivity of immunized rabbit serum from cercariae of cercariae infected with normal cercariae infected with ultraviolet light and different developmental stages. RESULTS AND CONCLUSION SDS-PAGE showed similarities and differences between immature egg antigen, mature egg antigen, cercariae antigen, male adult antigen and female adult antigen protein. Indirect ELISA tests showed that weak cercaria immune sera also induced high titer antibody levels in the host. A total of 23 kinds of antigenic molecules with immunological activity were screened from the immunized sera of weak cercariae. The molecular weights of them were 24.5kDa, 28kDa, 36kDa, 43kDa, 44kDa, 48kDa, 54kDa, 58.5kDa, 6 0 kDa, 62 kDa, 66 kDa, 68 kDa, 70 kDa, 75 kDa, 79 kDa, 85 kDa, 87 kDa, 91 kDa, 95 kDa, 97 kDa, 105 kDa, 108 kDa, 110 kDa. Among them, 11 were immature eggs and mature egg antigen, 4 were cercariae antigen molecules, 3 were male adult antigenic molecules and 6 were female adult antigenic molecules, suggesting that these antigenic molecules may be important for anti-Japanese schistosomiasis vaccine candidate molecules target