汉滩病毒M和S基因不同拼接方式嵌合基因免疫效果的研究  

Studies on Genetic Immunization of the Chimeric Genes of Hantaan virus M and S Segments that Spliced by Different Ways

作　　者：张芳琳[1] 徐志凯[1] 罗雯[1] 阎岩[1] 吴兴安[1] 刘勇[1] 白文涛[1] 赵茜[1] 王海涛[1] 

机构地区：[1]第四军医大学微生物学教研室,陕西西安710032

出　　处：《中国病毒学》2003年第1期14-17,共4页Virologica Sinica

基　　金：国家自然科学基金资助项目(30070686);国家教育部骨干教师资助计划资助项目

摘　　要：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。Recombinant euckaryotic expression vectors pcDNA3.1-G2S0.7 and pcDNA3.l-SO.7G2 were constructed by cloning chimeric genes containing G2 fragment of M segment and 0.7Kb fragment of S segment into pcDNAS.1 (+). Then BALB/c mice were vaccinated by the two vectors. ELISA results showed that both the vectors could induce specific antibodies against NP and GP. The specific antibody liters stimulated by pcDNA3.1-G2S0.7 were obviously higher that that of pcDNA3.l-SO.7G2. MTT results showed that the stimulation indexes of splenocytes of pcDNA3.1-G2S0.7 to NP and GP were significantly higher than that of control. However, that of pcDNA3.1-S0.7G2 were equal to that of control. It is suggested that the chimeric genes of Hantaan virus containing G2 fragment of M segment and 0.7Kb fragment of S segment could directly specific anti-Hantaan virus humoral immunity and cellular immunity in BALB/c mice. The different splicing ways could affect the immunity of chimeric genes.

关 键 词：汉滩病毒 M基因 S基因 嵌合基因 基因免疫 

分 类 号：R392[医药卫生—免疫学]