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目的 研究日本血吸虫中国大陆株 2 3k Da膜蛋白 (Sj C2 3) DNA疫苗诱导 BAL B/ c小鼠免疫保护作用以及 IL- 12的免疫增强效果。方法 5 6只 BAL B/ c雌性小鼠随机分为 A、B、C、D4组 ,每组 14只。 A组 (对照组 ) ,于每鼠两腿股四头肌注射 10 0 μg pc DNA3.1质粒 DNA;B组 (Sj C2 3组 ) ,注射 10 0μg pc DNA 3.1- Sj C 2 3质粒 DNA;C组 (Sj C 2 3+IL - 12 DNA组 ) ,注射 10 0μg pc D-NA3.1- Sj C 2 3、10 0μg pc DNA 3.1- p35、10 0μg pc DNA3.1- p40质粒 DNA的混合液 ;D组 (Sj C2 3+r IL- 12蛋白组 ) ,免疫剂量同 Sj C2 3组。共免疫 3次 ,每次免疫间隔 2周 ,剂量和方法同第 1次。于末次免疫后 1个月 ,每鼠攻击感染 (4 5± 2 )条日本血吸虫中国大陆株尾蚴。但 D组的小鼠在攻击感染当天 ,及感染后第 2、4、6天经腹腔注射 0 .2 μg/鼠 r IL- 12蛋白。攻击后 45 d剖杀小鼠 ,计数成虫及肝内虫卵数。于末次免疫后 2周对照组及 Sj C2 3组用 51 Cr释放法测定 Sj C 2 3介导的特异性细胞毒作用 ;用脾细胞培养法检测经重组日本血吸虫 2 3k Da膜蛋白亲水区大片段融合蛋白 (GST- HD)刺激后 ,在免疫后及攻击后小鼠脾细胞 IL - 2、IL - 4、IL - 10和 IFN -γ的水平。分别于攻击前后 2周经小鼠尾静脉采血 ,用 EL I
Objective To study the protective effect of Sj C2 3 DNA vaccine against schistosoma japonicum in Chinese mainland strain on immune protection and immune enhancement of BALB / c mice. Methods Sixty BAL B / c female mice were randomly divided into groups A, B, C and D4, with 14 mice in each group. Group A (control group) received 10 0 μg pcDNA3.1 plasmid DNA into each quadriceps quadriceps muscle. Group B (Sj C2 3 group) received 100 μg pc DNA 3.1-Sj C 2 3 plasmid DNA; Group C (Sj C 2 3 + IL - 12 DNA group), injection of 10 0μg pc D-NA3.1-Sj C 2 3,10 0μg pc DNA 3.1- p35,10 0μg pcDNA3.1-p40 plasmid DNA Liquid; group D (Sj C2 3 + r IL-12 protein group) and immunized dose with Sj C2 3 group. Immunization 3 times, each immunization interval of 2 weeks, the same dose and method with the first. One month after the last immunization, each rat challenge infected (45 ± 2) S. japonicum cercariae in China. However, mice in group D were injected intraperitoneally with 0.2 g / mouse rIL-12 protein on the day of challenge infection and on days 2, 4 and 6 of infection. Forty-five days after challenge, mice were sacrificed and the number of adult and intrahepatic egg masses was counted. At 2 weeks after the last immunization, the control group and Sj C2 3 group were treated with 51 Cr release assay Sj C 2 3-mediated specific cytotoxicity; Spleen cell culture was used to detect the hydrophilic region of the 23 kDa membrane protein of Schistosoma japonicum The levels of IL - 2, IL - 4, IL - 10 and IFN - γ in splenocytes of mice after immunization and challenged with GST - HD were measured. Blood was collected from the tail vein of mice 2 weeks before and after challenge, respectively, using EL I