日本血吸虫重组线粒体相关蛋白免疫学活性鉴定

分子筛进一步纯化并复性的融合蛋白均能刺激家兔产生特异性抗体 ,粗提包涵体蛋白免疫家兔 ,血清抗体滴度为 1∶2 5 6 0 0 ,复性蛋白及经分子筛纯化的蛋白免疫的家兔血清抗体滴度均为 1∶5 12 0 0 ,Westernblot结果显示 ,粗提包涵体蛋白、复性蛋白及经过分子筛进一步纯化并复性的蛋白均能被重感染兔血清和rSj338/2 6GST特异性免疫兔血清所识别。攻击实验中 ,粗提包涵体蛋白和经分子筛纯化的蛋白分别可诱导小鼠产生 2 7.8% (P <0 .0 1)和 30 .4 % (P <0 .0 1)的减虫率 ,4 0 .4 % (P <0 .0 1)和 4 3.5 % (P <0 .0 1)肝总减卵率。粗提包涵体蛋白中rSj338和经分子筛纯化的蛋白中rSj338分别可诱导小鼠产生 13.9% (P <0 .0 5 )和 2 0 .0 % (P <0 .0 5 )的减虫率 ,30 .0 % (P <0 .0 1)和 4 1.7% (P <0 .0 1)肝总减卵率。结论　获得的日本血吸虫线粒体相关的重组融合蛋白(rSj338/2 6GST)具有良好的免疫学活性 ,重组融合蛋白 (rSj338/2 6GST)及rSj338均可诱导宿主抗血吸虫感染的部分保护力。

Study on the immunogenicity of Schistosoma japonicum mitochondria related protein

Objective To identify the immunogenicity and the potential of using Schistosoma japonicum mitochondria related protein(rSj338) as a candidate vaccine antigen for Schistosomiasis japonica . Methods The expressed recombinant fusion protein(rSj338/26GST) was purified and recognized by S.japonicum heavily infected rabbit sera and rSj338/26GST immunized rabbit sera by Western blotting. The inclusion bodies,the revivified protein and the purified protein from S 12 gel filtration were injected twice into rabbits to produce anti rSj338/26GST antibody which titer was then measured by dot ELISA. Balb/c mice were immunized with the inclusion bodies and the purified protein from S 12 gel filtration and challenged with S. japonicum cercariae to identify the protective immunity produced by rSj338/26GST. Results The inclusion bodies, the revivified protein and the purified protein from S 12 gel filtration could stimulate the rabbits to produce high level of anti rSj338/26GST antibodies and could be recognized by S. japonicum heavily infected rabbit sera and rSj338/26GST immunized rabbit sera. In Balb/c mice, the inclusion bodies could induce 27.8%( P <0.01)worm reduction and 40.4%( P <0.01)total egg reduction in liver; the purified protein from S 12 gel filtration could induce 30.4% ( P <0.01)worm reduction and 43.5%( P <0.01)total egg reduction in liver. The rSj338 in the purified fusion protein could induce 20% worm reduction ( P <0.01).Conclusions The recombinant fusion protein (rSj338/26GST) and rSj338 possess immunogenicity in stimulating rabbits and mice to produce high level of specific antibody and have protective potential capable of being use as a candidate vaccine antigen against S. japonicum infection.