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目的 :观察本室所构建日本血吸虫大陆株 31kDa组织蛋白BDNA疫苗在BALB/c小鼠体内的不同免疫途径的免疫保护效果。方法 :用纯化的空白质粒载体VR1 0 1 2、重组质粒VR1 0 1 2 Sj31免疫BALB/c鼠 ,将 36只 6～ 8周龄BALB/c鼠随机分为 3组 ,每组 1 2只 ,对照组 (A组 )肌肉注射空白质粒载体VR1 0 1 2 ,实验组 (B组 )皮下注射重组质粒VR1 0 1 2 Sj31 ,C组肌肉注射重组质粒VR1 0 1 2 Sj31。质粒剂量均为 1 0 0 μg ,每隔 2w免疫 1次 ,共免疫 3次 ,第 3次免疫后第 2 1d ,每只鼠经腹部感染 1 0条尾蚴 ,4 2d后剖杀计数各小鼠成虫数和每克肝卵数 ,观察诱导产生的减虫和减卵效果 ,并用ELISA分析小鼠血清中的抗体。结果 :ELISA分析表明 ,第 3次免疫后小鼠出现特异性IgG抗体。与空白质粒对照组比较 ,2个实验组的减虫率分别为 1 5 .0 % (P >0 .0 5 )和 2 5 .0 % (P <0 .0 5 ) ,减卵率分别为 38.2 2 %和 5 4.90 % (P <0 .0 0 1 )。与皮下免疫组相比 ,VR1 0 1 2 /Sj31肌肉免疫组的减卵率分别为 2 6 .99% (P <0 .0 0 1 )。结论 :DNA疫苗VR1 0 1 2 Sj31能诱导小鼠产生一定水平的抗日本血吸虫感染保护作用 ,肌肉注射的保护效果大于皮下注射。
OBJECTIVE: To observe the immunoprotective effect of 31kDa cathepsin B DNA vaccine of Schistosoma japonicum constructed in this study in BALB / c mice. Methods: BALB / c mice were immunized with the purified plasmid vector VR1 0 1 2 and the recombinant plasmid VR1 0 1 2 Sj31. Thirty-six BALB / c mice aged 6-8 weeks were randomly divided into 3 groups (n = 12 each) The control group (group A) was intramuscularly injected with plasmid vector VR1 0 1 2, the experimental group (group B) was subcutaneously injected with the recombinant plasmid VR1 0 1 2 Sj31, and the group C was intramuscularly injected with the recombinant plasmid VR1 0 1 2 Sj31. The plasmid dose was 100 μg, immunized once every 2 weeks and co-immunized three times. On the 21st day after the third immunization, 10 cercariae were infected in the abdomen, and the mice were sacrificed after 42 days Number of adults and number of liver cells per gram, the induced reduction and ovulation reduction effects were observed, and the antibody in the mouse serum was analyzed by ELISA. Results: ELISA analysis showed that the mice developed specific IgG antibodies after the third immunization. Compared with the blank plasmid control group, the worm reduction rates of the two experimental groups were 15.0% (P> 0.05) and 25.0% (P <0.05), respectively. The egg reduction rates were 38.2 2% and 5 4.90% (P <0. 001). Compared with the subcutaneous immunization group, the egg reduction rates of VR1 0 1 2 / Sj31 muscle immunization group were respectively 26.99% (P <0.001). Conclusion: The DNA vaccine VR1 0 1 2 Sj31 can induce mice to produce a certain level of protection against Schistosoma japonicum infection. The intramuscular injection is more effective than subcutaneous injection.