日本血吸虫多价核酸疫苗的构建及免疫保护试验  

Construction and immunoprotection of Schistosoma japonicum multivalent DNA vaccine in mice

作　　者：魏峰[1,2] 翟羽佳[1,2] 刘全[1] 高胜岩[2] 门静涛[1] 商立民[1] 林矫矫[3] 傅志强[3] 朱兴全[4] 

机构地区：[1]军事医学科学院军事兽医研究所,吉林长春130062 [2]吉林农业大学生命科学学院,吉林长春130118 [3]中国农业科学院上海兽医研究所,上海200232 [4]华南农业大学兽医学院,广东广州510642

出　　处：《中国兽医学报》2009年第8期1013-1016,共4页Chinese Journal of Veterinary Science

基　　金：国家科技支撑计划资助项目(2006BAD06A09);国家"973"计划资助项目(2007CB513104)

摘　　要：利用PCR技术获得日本血吸虫GST抗原基因,通过分子克隆技术将GST、FABP基因克隆至pVAX1载体,并将其克隆至含有Sj23基因表达核的pIRESneo载体,构建重组质粒pVAX-GST、pVAX-GST-FABP、pIRESneo-Sj23和pIRESneo-GST-FABP-Sj23。将50只昆明小鼠随机分成5组,每组10只,分别以肌肉注射质粒pIRESneo-GST-FABP-Sj23、pIRESneo-Sj23、pVAX-GST、pVAX-GST-FABP和空白质粒pIRESneo,50μg/只,免疫2次,每次间隔21 d。测定小鼠免疫前后血清抗体水平,30 d经腹部皮肤感染血吸虫尾蚴,45 d后剖杀小鼠,计数各组小鼠虫卵数及成虫数。结果表明,重组质粒pVAX-GST、pIRESneo-Sj23、pVAX-GST-FABP和pIRESneo-GST-FABP-Sj23均能诱导小鼠产生特异性的IgG抗体,pIRESneo-GST-FABP-Sj23免疫组所诱导的IgG水平最高;减虫率分别为26.1%、30.8%、33.2%和47.5%;减卵率分别为25.4%、45.0%、48.4%和69.8%。pIRESneo-GST-FABP-Sj23的减卵率和减虫率明显高于其他DNA疫苗。结果表明,日本血吸虫多价核酸疫苗能产生较强的免疫反应,并能获得较好的免疫保护效果。The DNA vaccines against Schistosoma japonicum pVAX-GST, pVAX-GST-FABP, plRESneo-Sj23 and pIRESneo-GST-FABP-Sj23 were constructed. GST ORF was obtained by PCR from pGEX-4T-1 and cloned into the expression vector pVAX1 to generated pVAX-GST. FABP obtained from pGEX-FABP was directionaly inserted into pVAX-GST to construct the recombinant multivalent DNA vaccine pVAX-GST-FABP,which was subcloned into the expression vector plRESneo containing Sj23 to construct recombinant multivalent plasmid plRESneo-GST- FABP-Sj23. Fifty mice were divided into five groups,which were immunized intramuscularly with plRESneo-GST- FABP-Sj23,plRESneo-Sj23, pVAX-GST, pVAX-GST-FABP and blank plasmid plRESneo, respectively. The immunized mice were challenged withS, japonicurn cercariae 30 days after last vaccination. The antibody levels were determined by ELISA. At day 45 after challenge, all mice were sacrificed and perfused, the numbers of worms and eggs were counted. The results showed that pVAX-GST, plRESneo-Sj23, pVAX-GST-FABP and plRESneo-GST-FABP- Sj23 could induce the production of the specific antibody, the level in the plRESneo-GST-FABP-Sj23 was higher than other groups. The worm reduction rate were 26.1%, 30.8 %, 33.2 % and 47.5 %, respectively; the egg reduction rate were 25.4 %, 45.0 %,48.4 % and 69.8 %. Compared with other groups, the worm reduction rate and egg reduction rate of plRESneo-GST-FABP-Sj23 group was higher than other groups. It is concluded that the multivalent DNA vaccine expressing pIRESneo-GST-FABP-Si23 can provide certain protection against S. japonicum challenge in mice.

关 键 词：日本血吸虫 SJ23 脂肪酸结合蛋白 GST 多价DNA疫苗 

分 类 号：S852.7[农业科学—基础兽医学]