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目的观察日本血吸虫(大陆株)硫氧还蛋白重组抗原在小鼠诱导抗血吸虫感染的免疫保护作用。方法30只雌性C57BL/6小鼠随机分为3组,reSjcTrx免疫组:reSjcTrx(15mg/鼠)和ISA720佐剂乳化后采用小鼠相背部多点皮下注射,共免疫3次,间隔2周;ISA720佐剂对照组:小鼠仅注射ISA720佐剂和生理盐水;感染对照组不作任何处理。于末次免疫后3周,各组小鼠经腹部皮肤感染(30±1)条日本血吸虫尾蚴,感染后6周剖杀,门静脉灌注法收集成虫,计成虫数和每克肝组织虫卵数。在免疫前、攻击感染前和小鼠剖杀前分别采血并分离血清,用ELISA检测重组抗原特异性IgG抗体。并对reSjcTrx进行十二烷基磺酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)和蛋白质印迹(Westernblotting)分析。结果ELISA检测表明,reSjcTrx免疫组小鼠产生特异性IgG抗体应答,并诱导小鼠产生对攻击感染的减虫率和肝组织减卵率分别为22.8%和29.5%,与ISA720佐剂对照组和感染对照组相比,差异均有统计学意义(P﹤0.05)。SDS-PAGE和Westernblotting的结果表明,该重组抗原相对分子质量(Mr)约14000(含载体的6个氨基酸),可被日本血吸虫感染兔血清和reSjcTrx免疫小鼠血清所识别。结论reSjcTrx免疫小鼠可产生一定的抗血吸虫感染的保护作用。
Objective To observe the immunoprotective effect of thioredoxin recombinant antigen of Schistosoma japonicum (Chinese strain) on the induction of anti-schistosome infection in mice. Methods Thirty female C57BL / 6 mice were randomly divided into 3 groups: reSjcTrx immunized group: reSjcTrx (15mg / mouse) and ISA720 adjuvant. ISA720 adjuvant control group: mice injected only with ISA720 adjuvant and saline; infected control group without any treatment. At 3 weeks after the last immunization, mice in each group were infected with cercariae of Schistosoma japonicum (30 ± 1) through the skin of the abdomen. The mice were sacrificed at 6 weeks after infection, and the number of adult worms and the number of eggs per gram of liver tissue were counted by portal vein perfusion. Before immunization, before infection and before the mice were sacrificed, serums were collected and sera were separated, and the recombinant antigen-specific IgG antibodies were detected by ELISA. ReSjcTrx was analyzed by SDS-PAGE and Western blotting. Results ELISA test showed that mice immunized with reSjcTrx produced specific IgG antibody response and induced mice to reduce their worm infestation rate and the rate of reduction of liver tissue by 22.8% and 29.5%, respectively. Compared with ISA720 adjuvant control group and Compared with the control group, the differences were statistically significant (P <0.05). SDS-PAGE and Western blotting results showed that the relative molecular mass of the recombinant antigen (Mr) was about 14000 (containing 6 amino acids of the vector), which was recognized by the sera of rabbits infected with Schistosoma japonicum and the mice immunized with reSjcTrx. Conclusion The reSjcTrx immunized mice can produce certain protective effect against schistosomiasis infection.