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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疫苗诱导猪的保护性免疫作用。方法 构建并大量制备 pc DNA3.1- Sj C2 3质粒 DNA和 pc DNA3.1- p35 ,pc DNA3.1- p40(小鼠 IL - 12的 2个亚单位 )的 DNA疫苗。 30头 2 .5月龄的上海松江本地猪 (阉猪 )分为 A、B、C3组 ,每组 10头。 A组 (Sj C2 3组 )于每头猪两侧臀部肌肉注射 5 0 0 μg pc DNA3.1- Sj C2 3质粒 DNA,B组 (Sj C2 3+IL- 12组 ) ,于每头猪肌肉注射 5 0 0 μg pc DNA3.1- Sj C2 3DNA及 5 0 0 μg pc DNA3.1- p35和 5 0 0μg pc DNA 3.1- p40的混和质粒 DNA ;C组 (对照组 )每头猪肌肉注射 5 0 0μg pc DNA3.1质粒DNA。共免疫 3次 ,每次间隔 3周。末次免疫后 30 d,每猪经背部皮肤攻击感染日本血吸虫尾蚴 6 0 0条。攻击后 45 d剖杀 ,经胸主动脉灌冲 ,并从门静脉收集计算成虫。从肝脏的同一部位切下小块肝组织 ,置 5 % KOH内消化后 ,计算每克肝组织虫卵数 (EPG)。比较各组间减虫率、减雌率和减卵率。在免疫前 ,末次免疫后和攻击后从猪耳静脉采血 ,分离血清 ,用 EL ISA检测抗 Sj C2 3抗体。结果 Sj C2 3组与 Sj C2 3+IL - 12组与质粒对照组相比 ,分别获得 2 9.2 %和 5 8.6 %的减虫率、5 0 .8%和5 8.8%的减雌率以及 48.2 %和 5 6 .4%
Objective To study the protective immunity of porcine Schistosoma japonicum induced by DNA vaccine of 23 kDa membrane protein (Sj C2 3). Methods DNA vaccines of pc DNA3.1-Sj C2 3 plasmid DNA and pcDNA3.1- p35, pc DNA3.1-p40 (2 subunits of mouse IL-12) were constructed and extensively prepared. A total of 30 2.5-month-old Shanghai Songjiang local pigs (barrows) were divided into groups A, B and C3, 10 heads each. Group A (Sj C2 3) injected 500 μg pcDNA3.1-Sj C2 3 plasmid DNA into the buttocks of each pig at the buttocks, and group B (Sj C2 3 + IL-12) The mixed plasmid DNA of 500 μg pc DNA3.1-Sj C2 3DNA and 500 μg pc DNA3.1-p35 and 500 μg pc DNA 3.1-p40 was injected. Group C (control group) was intramuscularly injected with 5 0 0 pcDNA3.1 plasmid DNA. A total of 3 immunizations, each interval of 3 weeks. On the 30th day after the last immunization, 600 pigs were infected with S. japonicum cercarla by attacking the skin of the back. Forty-five days after challenge, the animals were sacrificed, infused via the thoracic aorta, and adult worms collected from the portal vein. Small pieces of liver tissue were cut from the same part of the liver, and the number of eggs per gram of liver tissue (EPG) was calculated after internal digestion with 5% KOH. Compare the worm reduction rate, decrease the female rate and reduce the egg reduction rate among the groups. Blood was collected from porcine ear veins before, after, and after the last immunization, and sera were isolated and anti-Sj C2 3 antibodies were detected by EL ISA. Results Compared with plasmid control group, Sj C2 3 group and Sj C2 3 + IL - 12 group achieved worm reduction rates of 22.6% and 5.8%, reduction rates of 0.58% and 5.88%, and 48.2% % And 56.4%