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目的 探讨日本血吸虫中国大陆株磷酸丙糖异构酶 (Sj CTPI) DNA疫苗诱导猪的保护性免疫作用。方法 分别构建并制备 pc DNA 3.1- Sj CTPI DNA疫苗及 pc DNA3.1- P35和 P40 (IL - 12的两个亚单位 )的 DNA疫苗。取上海松江本地猪 (自幼阉割 ) 30头 ,分为 A、B、C 3组 ,每组 10头。 A组 (TPI组 )每头猪于两侧臂部肌肉注射 5 0 0 μg Sj CTPI DNA疫苗 ;B组 (TPI+IL- 12组 )每头猪肌肉注射 5 0 0 μg Sj CTPI DNA及 5 0 0 μg P35 DNA疫苗、5 0 0 μg P40 DNA疫苗。C组 (对照组 )每头猪肌肉注射 5 0 0 μg pc DNA3.1质粒 DNA。共免疫 3次 ,每次间歇 3周。末次免疫后 30 d,每猪攻击 6 0 0条日本血吸虫尾蚴 (背部贴片法 )。攻击后 45 d剖杀 ,经胸主动脉灌注 ,并从门静脉处收集成虫计数。从肝脏上同一部位切下小块肝组织 ,置 5 % KOH内溶解 ,后计算每克虫卵数。比较各组间的减虫率、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法测抗体。结果 TPI组和 TPI+IL- 12组与质粒对照组相比 ,分别获得 48.3%和 46 .2 %的减虫率 ,5 3.6 %和5 9.6 %的减雌率 ,49.4%和 6 5 .8%的减卵率。TPI组的 10头猪中有 6头在免疫后可检出抗 r Sj CTPI抗体 ,TPI+IL- 12组 10头猪中有 5头在免疫
Objective To investigate the protective immunity induced by Sj CTPI DNA vaccine of Schistosoma japonicum in Chinese mainland. Methods The DNA vaccine of pcDNA3.1-Sj CTPI DNA vaccine and pcDNA3.1-P35 and P40 (two subunits of IL-12) were constructed and prepared respectively. Take the Shanghai Songjiang local pigs (castration since childhood) 30, divided into A, B, C 3 groups of 10. Group A (TPI) piglets were injected intramuscularly with 500 μg Sj CTPI DNA vaccine in each arm of each group; Group S (TPI + IL-12) were injected intramuscularly with 500 μg Sj CTPI DNA and 50 0 μg P35 DNA vaccine, 500 μg P40 DNA vaccine. Group C (control group) were intramuscularly injected with 500 μg of pc DNA3.1 plasmid DNA. Immunization 3 times, each intermittent 3 weeks. On the 30th day after the last immunization, 600 pigs per kilogram were challenged with cercariae of Schistosoma japonicum (back patch method). Forty-five days after challenge, animals were sacrificed, perfused through the thoracic aorta, and adult worms collected from the portal vein. Small pieces of liver tissue were excised from the same area on the liver, lysed in 5% KOH, and the number of eggs per gram was calculated. Compare the worm reduction rate among groups, reduce the female rate and decrease the egg rate. Blood was collected from the pig’s ear veins before, after, and after the last immunization, the serum was separated and antibodies were measured by EL ISA. Results Compared with the plasmid control group, TPI group and TPI + IL-12 group achieved worm reduction rates of 48.3% and 46.2%, reduction rates of 53.6% and 59.6%, 49.4% and 65.8% respectively % Of the egg reduction rate. Anti-r Sj CTPI antibodies were detected in 6 out of 10 pigs in the TPI group and 5 in 10 pigs in the TPI + IL-12 group