日本血吸虫GST、Sj23及GST-Sj23 RNA颗粒抗原的制备及表达研究

目的制备日本血吸虫自杀性RNA颗粒抗原,并研究其在真核细胞内的表达,以期探讨新型抗原的研制方法,为制备抗血吸虫疫苗奠定基础。方法将编码日本血吸虫主要抗原GST、Sj23及编码两种抗原的嵌合基因(GST-Sj23)分别克隆到自杀性RNA疫苗载体SFV3.spider内,以体外转录法制备编码上述三种抗原的mRNA。同法合成编码SFV病毒表膜蛋白质(S蛋白质和C蛋白质)的mRNA,然后将编码上述三种抗原的mRNA分别与编码病毒膜蛋白质的mRNA混合,以电穿孔法导入BHK21细胞,制备含有日本血吸虫GST、Sj23抗原及GST-Sj23嵌合体抗原mRNA的假病毒颗粒。最后,对制备出的假病毒颗粒进行真核细胞感染实验,以间接免疫荧光法验证上述抗原的真核表达情况。结果三种RNA颗粒对BHK21细胞均具有很好的感染性,所编码的蛋白质在BHK21细胞内鉴定高效表达。

Generation of GST, Sj23 and GST-Sj23 recombinant RNA particle antigens of Schistosoma japonicum and their expressions in mammalian cells

To generate the recombinant suicide RNA-based antigens of Schistosoma japonicurn and their expressions in mammalian cells, genes encoding two schistosomal key antigens （GST and Sj23） and a hybrid gene GST-Sj23 were cloned into Semliki forest virus （SFV） vaccine vector and the recombinant suicidal RNA virus particles containing the schistosomal mRNA were packaged by transforming BHK21 cells with mRNA encoding the viral structural proteins, together with mRNA of either GST, Sj23 or the hybrid proteins. RNA particles were harvested and tested for their ability to deliver the antigen coding mRNA into mammalian cells. Immuno-fluorescence assay was used to confirm the expression of the recombinant proteins after virus in-fection in BHK21 cells. It was demonstrated that the parasitic antigens could be expressed and displayed on the infected cell surface after addition of the recombinant virus particles into culture of BHK21 cells.