汉坦病毒嵌合基因G2S0.7在昆虫细胞中融合表达产物的免疫学研究

Studied on immunological properties of Hantaan virus fusion protein G2S0.7

在前期工作基础上,对汉坦病毒糖蛋白(GP)和核蛋白(NP)的嵌合基因G2S0.7表达产物的免疫学特性进行进一步的研究。将汉坦病毒含有G2S0.7嵌合基因的重组杆状病毒在昆虫细胞中融合表达并免疫BALB/c小鼠,用ELISA、微量细胞培养中和实验及淋巴细胞增殖实验检测免疫应答效果,以研究嵌合基因的免疫效果。结果表明,用该融合蛋白免疫小鼠,可诱导产生抗汉坦病毒NP及GP特异性的抗体,抗体效价分别为1∶3200及1∶200;同时融合蛋白还可刺激机体产生低水平的中和抗体和明确的淋巴细胞增殖反应。说明汉坦病毒G2S0.7嵌合基因表达的融合蛋白既可刺激机体产生特异性的抗汉坦病毒体液免疫应答,也可刺激机体产生特异性的细胞免疫应答,为进一步进行汉坦病毒基因工程疫苗的研究奠定了实验基础。

To investigate the immunological properties of Hantaan virus fusion protein G2S0.7.Fusion protein G2S0.7was expressed in insect cells and BALB/c mice were immunized by it.The immune response was tested by ELISA,microcell-culture neutralizing experiment and T lymphocyte proliferation test(MTT assay).The results showed that the fusion protein of sf9cells infected by Bac-G2S0.7can stimulate high-titer combining antibodies against NP(1∶3200)and GP(1∶200)respectively detected by ELISA,which suggested both GP and NP immune response were acti-vated.The low-titer neutralization antibody reaction and clear lymphocyte proliferation were detected simultaneity.It is suggested that the chimeric gene of Hantaan virus can directly stimulate BALB/c mice to produce specific hu-moral immunity and cellular immunity in against hantaan virus.It would provide theoretical and experimental evi-dences for the development of genetic engineering vaccine efficient against HFRS.