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目的 观察本课题组所构建日本血吸虫大陆株二价 DNA疫苗 VR10 12 - Sj GST- Sj31和VR10 12 - Sj GST- Sj32的免疫保护效果。方法 用纯化质粒免疫昆明鼠 :6 0只小鼠分为 5组 ,2个对照组的小鼠分别于股四头肌注射生理盐水 10 0μl或空质粒 VR10 12 10 0μg,3个实验组的小鼠则同法分别注射 VR10 12 - Sj GST、VR10 12 - Sj GST- Sj31和 VR10 12 - Sj GST- Sj32各 10 0 μg,每隔 3周免疫1次 ,共免疫 3次 ,以间接免疫荧光法观察 VR10 12 - Sj GST- Sj31、VR10 12 - Sj GST- Sj32在肌肉组织中的表达后 ,每只鼠经腹部感染 10条尾蚴 ,4 5 d后剖杀计数各小鼠成虫数及肝卵数 ,并用 EL ISA分析小鼠血清中的抗体。结果 EL ISA分析表明 ,第 3次免疫后小鼠出现特异性 Ig G抗体。与生理盐水组比较 ,3个实验组的减虫率分别为 33.90 %、33.90 %及 2 7.14 % (P <0 .0 1) ,减卵率分别为6 1.86 %、74 .88%及 6 8.87% (均为 P<0 .0 1) ,每雌肝减卵率分别为 4 4 .6 7%、5 9.4 0 %、5 4 .32 % (P<0 .0 1)。与 VR10 12 - Sj GST组相比 ,VR10 12 - Sj GST- Sj31和 VR10 12 - Sj GST- Sj32组的减卵率分别为34.19% (P<0 .0 1)、18.5 1% (P<0 .0 1) ,每雌肝减卵率为 2 6 .6 2 %、17.4 6 % (P<0 .0 1)。结论 DNA疫苗 VR10 12 -
Objective To observe the immunoprotective effect of the bivalent DNA vaccine VR10 12 - Sj GST-Sj31 and VR10 12 - Sj GST-Sj32 constructed by our team. Methods Kunming mice were immunized with the purified plasmids: 60 mice were divided into 5 groups. The mice in 2 control groups were injected with 10 0μl normal saline or empty plasmid VR10 12 10 0μg in quadriceps femoris, respectively Rats were injected with the same method of VR10 12 - Sj GST, VR10 12 - Sj GST-Sj31 and VR10 12 - Sj GST-Sj32 each 10 0 μg, immunized once every 3 weeks, three times immunized with indirect immunofluorescence After the expression of VR10 12 - Sj GST - Sj31, VR10 12 - Sj GST - Sj32 in muscle tissue, 10 cercariae were infected in the abdomen and the number of adult mice and the number of liver eggs , And antibodies in mouse serum were analyzed by EL ISA. Results EL ISA analysis showed that specific immunoglobulin G (IgG) antibodies appeared after the third immunization. Compared with the saline group, the worm reduction rates of the three experimental groups were 33.90%, 33.90% and 21.74% (P <0.01) respectively, and the egg reduction rates were 6 1.86%, 74.88% and 6 8.87 % (P <0. 01). The rate of egg loss per female was 44.76%, 54.04% and 54.32% (P <0.01), respectively. Compared with VR10 12 - Sj GST group, the egg reduction rates of VR10 12 - Sj GST - Sj31 and VR10 12 - Sj GST - Sj32 were 34.19% (P <0.01) and 18.5% (P <0) .0 1), and the rate of egg reduction per female liver was 26.62% and 17.4% (P <0.01). Conclusion DNA Vaccine VR10 12 -