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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 GST antigen gene of Schistosoma japonicum was obtained by PCR. The GST and FABP genes were cloned into pVAX1 vector by molecular cloning technology and cloned into pIRESneo vector containing Sj23 gene expression core to construct recombinant plasmids pVAX-GST, pVAX-GST-FABP , PIRESneo-Sj23 and pIRESneo-GST-FABP-Sj23. Fifty Kunming mice were randomly divided into five groups of 10 mice. The plasmids pIRESneo-GST-FABP-Sj23, pIRESneo-Sj23, pVAX-GST, pVAX-GST-FABP and blank plasmid pIRESneo were injected intramuscularly , Immunized 2 times, each interval of 21 d. The level of serum antibody was measured before and after immunization. After 30 days, the cercariae of Schistosoma japonicum were infected by abdominal skin. After 45 days, the mice were sacrificed and the numbers of eggs and number of adults in each group were counted. The results showed that the recombinant plasmids pVAX-GST, pIRESneo-Sj23, pVAX-GST-FABP and pIRESneo-GST-FABP-Sj23 all induced specific IgG antibodies in mice. The immunogenicity induced by pIRESneo-GST-FABP-Sj23 IgG was the highest. The worm reduction rates were 26.1%, 30.8%, 33.2% and 47.5%, respectively. The egg reduction rates were 25.4%, 45.0%, 48.4% and 69.8%, respectively. The ovulation reduction and worm reduction rates of pIRESneo-GST-FABP-Sj23 were significantly higher than those of other DNA vaccines. The results showed that the multivalent nucleic acid vaccine of Schistosoma japonicum could produce a strong immune response and obtain a better immune protection effect.