日本血吸虫Calpain疫苗候选分子抗感染的免疫机制  

Immunoprotective Mechanism of Calpain as Vaccine Candidate against Schistosoma japonicum Infection

作　　者：张仁利[1] 黄达娜[1] 高世同[1] 耿艺介[1] 吴少庭[1] 胡章立[2] 张顺祥[1] 程锦泉[1] 

机构地区：[1]深圳市疾病预防控制中心,深圳518020 [2]深圳大学生命科学院,深圳518060

出　　处：《热带医学杂志》2006年第12期1260-1263,共4页Journal of Tropical Medicine

基　　金：国家教委留学回国人员科研启动基金(No.2001[345])

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BALB/c小鼠,制备抗Calpain血清,抗Calpain血清与机械转化的日本血吸虫童虫和来自于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粘附及对童虫的杀伤效果;RT-PCR分析Calpain抗原免疫BALB/c小鼠一氧化氮激酶(iNos.eNos.nNos)mRNA的表达;ELISA分析免疫小鼠体外培养脾细胞产生的细胞因子IFN-γ和IL-4。结果重组Calpain抗原免疫小鼠后,产生了一个极高的抗Calpain特异性抗体,此抗体介导了同种鼠的嗜酸性粒细胞对日本血吸虫童虫的粘附,37℃、5%CO2培养48h后,33.8%的日本血吸虫童虫被杀死,与控制组14.5%的自然死亡率相比有一个显著性的增高(P<0.01);RT-PCR测定重组Calpain抗原免疫1周小鼠iNosmRNA的表达显著性的增强,培养24h脾细胞上清中IFN-γ浓度高于对照组。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,Calpain抗原能够诱导IFN-γ的产生,提示Calpain诱导Th1的免疫应答反应来抗日本血吸虫的感染。Objective To evaluate Calpain, a calcium-activated neutral proteinase as a vaccine candidate against Schistosoma japonicum infection. Method Anti-calpain sera was prepared by immunizing BALB/c mice with recombinant calpain. Anti-calpain sera, schistosomula of S-japonicum and mouse eosinophils were culture at 37℃, 5% CO,. for 48 hours, and the schistosomulacidal was determined, mRNA of iNos. nNos and enos were decteced by RT-PCR in immunized BALB/c mice, and concentrations of IFN-γ and IL-4 in spleen cell of culture of immunizing mice were analyzed by ELISA. Result Mice immunized by recombinant calpain produced a high level of anti-calpain IgG. Immunoblotting analysis showed mouse anti-calpain serum could specifically binds to recombinant calpain. Ninety six percent of schistosomula were covered by cells when incubated with mouse eosinophils, and A significant number of death of schistosomula was observed at 48 hours when incubated with mouse anti-serum and eosinophils as comparing with control serum and eosinophils（P 〈 0.01 ）. Both iNos mRNA level and concentration of IFN-γin immunized mice were significant increased. Conclusion Anti-calpain antibodies can induce antibody dependent cell-adherence and killing of the schistosomula, which suggested immunization with calpain of S. japonicum may cause Thl immune reaction due to IFN-γ increase in immunized mice.

关 键 词：日本血吸虫 免疫机理 细胞毒 CALPAIN TH1免疫应答 

分 类 号：R383.24[医药卫生—医学寄生虫学]