流感病毒NA—C3d融合基因在非复制型重组腺病毒中的表达及其免疫效果的研究  

Expression of NA of influenza virus and C3d fusion gene in replication-defective recombinant adenovirus and its immune efficacy analysis

作　　者：韩峰[1] 王璇[1] 王艳[1] 赵小东[1] 吴淑华[1] 

机构地区：[1]中国疾病预防控制中心病毒病预防控制所,北京102206

出　　处：《中华实验和临床病毒学杂志》2013年第1期5-7,共3页Chinese Journal of Experimental and Clinical Virology

摘　　要：目的构建表达融合基因NA—C3d的非复制型重组腺病毒并研究其免疫效果。方法将NA-C3d融合基因克隆到穿梭载体pAdTrack—CMV，与腺病毒DNA共转化E.coliBJ5183，经同源重组获得重组腺病毒DNA，将其转染293细胞获得重组腺病毒并检测其免疫效果。结果NA-C3d经PCR证实整合至腺病毒基因组中；重组病毒感染293细胞中经WsternBot检测到融合蛋白；重组病毒经滴鼻途径免疫小鼠，2次免疫后产生明显的免疫应答，血清IgG抗体滴度分别为1：1000和1：100000；经小剂量攻毒实验显示重组腺病毒保护率为100％。结论成功构建表达NA—C3d的非复制型重组腺病毒，该病毒可诱导较好的免疫效果。Objective To construct a replication-defective recombinant adenovirus expressing the fusion gene of neuraminidase （NA） gene in influenza virus A/FM/1/47 and C3d and to evaluate the induced immune efficacy. Methods NA-C3d was cloned into shutter vector pAdTrack-CMV, which was cotransformated with adenovirus DNA into E. coli BJ5183. The recombinant adenovirus gcnomic DNA was generated through homologieal recombination. The recombinant adenovirus was produced by transfecting 293 cell line with the genomic DNA and the induced immune efficacy in mice were analyzed. Results The integration of NA-C3d in tile adenovirus genomie DNA and its expression were confirmed by PCR and Western-Blot assays respectively. After intranasal immunization, the serum IgG was induced at a titer of 1： 1000 and 1：100 000 in BALB/c mice at primary and secondary immunization respectively. The vaccinated mice were completely survived when challenged with wide influenza virus. Conclusion recombinant adcnovirus expressing NA-C3d was successfully constructed and it could induce desired immune efficacy.

关 键 词：流感病毒A型 神经氨酸酶 佐剂 免疫 腺病毒科 

分 类 号：R3[医药卫生—基础医学]