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目的 研究日本血吸虫中国大陆株磷酸丙糖异构酶 (Sj CTPI) DNA疫苗诱导 BAL B/ c小鼠的保护性免疫作用。方法 将 39只 BAL B/ c小鼠随机分成 3组 :A组 (对照组 ) ,肌注 pc DNA3.1质粒 DNA10 0 μg/鼠 ;B组 (实验组 ) ,每鼠肌注 pc DNA3.1- Sj CTPI10 0 μg;C组 (加强组 ) ,每鼠肌注 pc DNA3.1- Sj CTPI10 0 μg及 pc DNA3.1- P35和 pc DNA3.1-P40的混合物 10 0 μg。每两周免疫 1次 ,共计 3次。末次免疫 4周后 ,每鼠用 45条尾蚴进行攻击 ,45 d后剖杀小鼠 ,计数成虫以及每鼠肝组织内虫卵数 ;于攻击前每组各取两只小鼠采用 51 Cr释放法测定 Sj CTPI介导的特异性细胞毒作用 ;EL ISA检测攻击前后的抗 TPI的抗体水平。 结果 用 EL ISA检测抗 TPI抗体的结果表明 ,攻击前 A组的 10份血清均为阴性 ,B组 5 / 10份血清出现弱阳性 ,C组也有 6 / 10份血清出现弱阳性反应。51 Cr释放法测定细胞毒活性结果表明 ,A、B、C组细胞毒活性分别为 9.1%、2 7.6 %和 5 4.4%。与对照组相比 ,实验组的减虫率、减卵率分别为 30 .2 %、5 2 .9% ;加强组的减虫率、减卵率分别为 32 .7%、47.0 %。 结论 进一步证实了 Sj CTPI DNA疫苗作为抗血吸虫病核酸疫苗的潜力。
Objective To study the protective immunity of BALB / c mice induced by Sj CTPI DNA vaccine of Chinese mainland Schistosoma japonicum. Methods Thirty-nine BALB / c mice were randomly divided into three groups: group A (control group), intramuscular pcDNA3.1 plasmid DNA of 100 μg / mouse, group B (experimental group), intramuscular pcDNA3.1 - Sj CTPI10 0 μg; Group C (enhanced group), intramuscular injection of pcDNA3.1-Sj CTPI10 0 μg per mouse and a mixture of pcDNA3.1- P35 and pcDNA3.1-P40 10 0 μg. Immunized once every two weeks for a total of 3 times. Four weeks after the last immunization, each mouse was challenged with 45 cercariae. After 45 days, the mice were killed and the number of eggs in the adult and liver tissues of each mouse was counted. Two mice from each group were challenged with 51 Cr Method to determine the specific cytotoxicity mediated by Sj CTPI. The levels of anti-TPI antibody before and after challenge were detected by ELISA. Results The results of ELISA assay of anti-TPI antibody showed that all the 10 serums in group A before challenge were negative, 5/10 serums in group B were weakly positive, and 6/10 serums in group C showed a weak positive reaction. The results of 51 Cr release assay showed that the cytotoxic activities of group A, B and C were 9.1%, 7.66% and 5.44%, respectively. Compared with the control group, the worm reduction rate and the oviposition rate of the experimental group were 30.2% and 52.2%, respectively. The worm reduction and reduction rates of the experimental group were 32.7% and 47.0%, respectively. The conclusion further confirms the potential of the Sj CTPI DNA vaccine as a DNA vaccine against schistosomiasis.