PbTIP多克隆抗体对恶性疟原虫生长抑制作用的初步研究

摘要

To preparepolyclonal anti-T cell immunomodulatory protein of Plasmodium berghei and exam effect of it on growth of Plasmodium falciparum.P&TIP gene was constructed in pET32a( + ) vector with a His tag and successfully expressed by E.Coli.The PbTlP-His fusion protein was characterized by SDS-PAGE and Western blots.The polyclonal PbTIP antibody was produced by BALB/c mouse immunized by purified PbTIP-His fusion protein, and the effect of it on growth of Plasmodium falciparum was determined by infection ratio.PbTIP-His fusion protein was successfully expressed in the form of inclusion bodies as evidenced by SDS-PAGE and western blot results.Anti-PbTIP sera were collected in mice enhanced immunized by PbTIP-His protein at a titer of 1:105, which inhibited growth of P.f parasites as evidenced by reduced infection ratio in RBC infected by Plasmodium falciparum.The construction and purification of PbTIP-His fusion protein together with poly-anti-PbTIP could be used for further investigation in Plasmodium berghei, especially for the further study of PbTIP in parasite-host relationship.%首先构建了伯氏疟原虫TIP样蛋白(以下简称PbTIP)的原核表达载体pET32a/PbTIP,在大肠杆菌中成功诱导表达了带His标签的PbTIP融合蛋白.SDS-PAGE分析显示该融合蛋白主要以包涵体形式表达.Western-Blot鉴定虽然证明该融合蛋白表达正确,但即便在变性条件下该蛋白的亲和层析纯化效果也不佳.以PbTIP融合蛋白经过初次和三次加强免疫获得的小鼠PbTIP多克隆抗体滴度达到1∶105以上.进一步的研究表明,小鼠PbTIP抗血清对体外培养恶性疟原虫的生长具有一定的抑制作用,这显示PbTIP多克隆抗体对恶性疟原虫感染具有一定的交叉保护作用.以上实验结果为后续PbTIP在疟原虫免疫抑制中的功能及其机制研究奠定了一定的基础.