论文部分内容阅读
目的在本室前期工作的基础上,进一步进行汉滩病毒M基因G2片段与S基因0.7Kb片段嵌合基因基因免疫的研究。方法构建汉滩病毒76-118株M基因G2片段与S基因5′端700bp片段的嵌合基因真核表达载体pcDNA3.1-G2S0.7。用该质粒免疫Balb/c小鼠,并用ELISA法及淋巴细胞增殖实验,检测基因免疫后的免疫应答效果。结果限制性内切酶鉴定结果表明真核表达载体的构建正确。用pcDNA3.1-G2S0.7直接免疫小鼠,可诱导产生抗汉滩病毒核蛋白NP及糖蛋白GP特异性的抗体,抗体效价分别为1200及180。淋巴细胞增殖实验表明,嵌合基因免疫小鼠脾细胞对NP及GP的增殖指数,均明显高于对照组。结论汉滩病毒M基因G2片段及S基因0.7kb片段的嵌合基因,既可刺激机体产生特异性的抗汉滩病毒体液免疫应答,也可刺激机体产生特异性的细胞免疫应答,本研究为进一步进行汉滩病毒基因疫苗的研究奠定了实验基础。
OBJECTIVE To further study the gene immunization of chimeric gene of Hantaan virus M gene G2 fragment and S gene 0.7Kb fragment on the basis of previous work in our hospital. Methods The eukaryotic expression vector pcDNA3.1-G2S0.7 of chimeric gene of Hantaan virus 76-118 M gene G2 fragment and S gene 5 ’end 700bp fragment was constructed. Balb / c mice were immunized with the plasmid and the effect of immune response after gene immunization was tested by ELISA and lymphocyte proliferation assay. Results The results of restriction endonuclease identification showed that the eukaryotic expression vector was constructed correctly. Direct immunization of mice with pcDNA3.1-G2S0.7 induced anti-Hantaan virus nucleoprotein NP and glycoprotein GP-specific antibodies with antibody titers of 1200 and 180, respectively. Lymphocyte proliferation experiments showed that the spleen cells of chimeric gene immunized mice NP and GP proliferation index were significantly higher than the control group. Conclusion The chimeric gene of Hantaan virus M gene G2 fragment and S gene 0.7kb fragment can not only stimulate the body to produce specific anti-Hantaan virus humoral immune response, but also stimulate the body to produce specific cellular immune response Further research on Hantaan virus gene vaccine laid the foundation for the experiment.