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本文在前期工作的基础上,构建了汉滩病毒76-118株M基因G2片段与S基因5’端0.7Kb片段的嵌合基因真核表达载体pcDNA3.1-G2S0.7及pcDNA3.1-S0.7G2;用该质粒免疫BALB/c小鼠。结果表明两种质粒免疫小鼠可同时诱导产生抗汉滩病毒核蛋白(NP)及糖蛋白(GP)特异性的抗体,且前者刺激产生的抗体效价明显高于后者。淋巴细胞增殖实验表明,pcDNA3.1-G2S0.7组免疫小鼠脾细胞时NP及GP的增殖指数均明显高于空载体对照组,而pcDNA3.1-S0.7G2组未检测到其淋巴细胞有明显的增殖。这说明汉滩病毒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的抗汉滩病毒体液免疫应答,也可刺激机体产生特异的细胞免疫应答。不同拼接方式对嵌合基因免疫效果有很大影响,嵌合基因G2S0.7这种拼接方式明显优于S0.7G2。
Based on the previous work, we constructed the eukaryotic expression vectors pcDNA3.1-G2S0.7 and pcDNA3.1-G2S0 of chimeric gene of G2 fragment of Hantavirus 76-118 M and 0.7Kb fragment of S gene 5 ’ S0.7G2; BALB / c mice were immunized with this plasmid. The results showed that both of the plasmid-immunized mice could simultaneously induce antibodies specific to NP and GP of Hantaan virus and the antibody titer produced by the former was significantly higher than the latter. Lymphocyte proliferation experiments showed that the proliferation index of NP and GP in spleen cells of mice immunized with pcDNA3.1-G2S0.7 were significantly higher than that of empty vector control group, while the pcDNA3.1-S0.7G2 group did not detect lymphocytes Significant proliferation. This indicates that the chimeric gene of the Hantaan virus M gene G2 fragment and the S gene 0.7Kb fragment can both stimulate the body to produce specific anti-Hantaan virus humoral immune response and stimulate the body to produce a specific cellular immune response. Different splicing methods have a great influence on the immunogenicity of chimeric genes. The splicing method of chimeric gene G2S0.7 is obviously better than that of S0.7G2.