日本血吸虫重组蛋白LHD-Sj23-GST诱导的细胞免疫应答和免疫保护效果  

Cellular immune response and immuno-protective efficacy induced by the recombinant protein LHD-Sj23-GST of Schistosoma japonicum

作　　者：朱珠[1] 傅志强[1] 洪炀[1] 韩艳辉[1,2] 张旻[1] 李东[1] 石耀军[1] 林矫矫[1] 

机构地区：[1]中国农业科学院上海兽医研究所,上海200241 [2]南京农业大学动物医学院,江苏南京210095

出　　处：《中国兽医科学》2013年第1期36-41,共6页Chinese Veterinary Science

基　　金：国家自然科学基金资助项目(31172315);公益性行业(农业)科研专项(200903036)

摘　　要：为探讨日本血吸虫重组蛋白LHD-Sj23-GST与3种不同类型佐剂(弗氏佐剂、ISA 206佐剂、ISA70M佐剂)配伍后免疫小鼠诱导的细胞免疫应答及免疫保护效果,应用流式细胞术检测并比较该重组蛋白配伍3种佐剂3次免疫后小鼠脾细胞中的CD4+、CD8+T细胞及细胞因子IFN-γ、IL-4、IL-10水平;3次免疫后小鼠人工攻击感染日本血吸虫尾蚴,6周后经肝门静脉灌注冲虫,计数各组平均虫体数并计算减虫率。与PBS对照组相比,LHD-Sj23-GST/FA和LHD-Sj23-GST/70M免疫组的CD4+T细胞显著升高(P<0.05),LHD-Sj23-GST/206免疫组未呈现明显变化;3种不同佐剂配伍LHD-Sj23-GST免疫组CD8+T细胞水平均显著降低(P<0.05);与PBS对照组相比,免疫组的IFN-γ及IL-10水平升高,而IL-4水平降低。LHD-Sj23-GST/FA、LHD-Sj23-GST/206和LHD-Sj23-GST/70M免疫组与PBS对照组相比,分别获得65.52%、51.72%和51.72%的减虫率;而相较于各自的佐剂对照组,则分别获得58.76%,26.31%,54.28%的减虫率。结果表明,用重组蛋白LHD-Sj23-GST配伍3种不同类型佐剂免疫小鼠,均刺激产生了偏向Th1型的细胞免疫应答,且都对小鼠诱导了较高的免疫保护作用。To investigate the cellular immune response and protective efficacy induced by the recombinant protein LHD-Sj23-GST（large hydrophilic domain of 23 ku antigen of Schistosoma japonicum fused with pGEX expression protein）,mice were vaccinated with the recombinant protein emulsified with three different adjuvants CFA/IFA,ISA 206 and ISA 70M.The percentage of CD4＋ and CD8＋ T cells,IFN-γ cells,IL-4 cells and IL-10 cells from splenocytes in all test groups were analyzed through flow cytometry.The mice were challenged with cercariae of S.japonicum and perfused at 6 weeks post infection,the worms were collected and counted,the worm reduction rates were calculated and the protective efficacies were evaluated.Flow cytometric analyses revealed that the percentage of CD4＋ T cells was significantly higher（P0.05） in mice from LHD-Sj23-GST/FA and LHD-Sj23-GST/70M groups,but not obviously changed in LHD-Sj23-GST/206 group,and the CD8＋ T cells were significantly lower（P 0.05） in mice from all the three LHD-Sj23-GST vaccinated groups when compared with the PBS control group.In addition,-the percent of Th1 cytokine-dependent IFN-γ cells as well as IL-10 cells were significantly higher,and Th2 cytokine-dependent IL-4 cells was significantly lower in the LHD-Sj23-GST-vaccinated mice.Animal vaccination showed that 65.52%,51.72% and 51.72% worm reduction rates（P0.05） were obtained in the LHD-Sj23-GST/FA,LHD-Sj23-GST/206 and LHD-Sj23-GST/70M groups respectively when compared with the PBS control group,while 58.76%,26.31% and 54.28% worm reduction rates（P 0.05） were observed when compared with each respective adjuvant control group.The results suggested that the recombinant protein LHD-Sj23-GST emulsified with the three different adjuvants induced significantly immuno-protective and obvious Th1 cellular immuno-response in BALB/c mice.

关 键 词：日本血吸虫 重组蛋白LHD-Sj23-GST 佐剂 细胞免疫应答 免疫保护效果 

分 类 号：S852.735[农业科学—基础兽医学]