含H5N1-HA基因重组腺病毒疫苗的构建及其诱导免疫应答的初步探讨

Study on the immunogenicity of adeno-vector vaccine against H5N1 influenza A virus

目的构建包含H5N1-HA基因的重组腺病毒疫苗并探讨其免疫效果。方法用Admax系统构建包含H5N1-HA基因的重组腺病毒疫苗，并用PCR、Western—Blot等方法对重组病毒疫苗进行鉴定；疫苗免疫小鼠后，通过H1实验和ELISPOT实验检测其体液免疫和细胞免疫反应，评价其免疫效果。结果成功得到了含有H5N1-HA基因的重组腺病毒疫苗；基因表达鉴定表明，HA基因能够在细胞中进行表达；血凝抑制实验结果显示小鼠产生的针对HA抗体滴度在1：320和1：640之间；ELISPOT结果显示实验组和对照组（PBS）相比斑点数量差异有统计学意义（P〈0．05），以上免疫结果表明重组腺病毒载体疫苗可以诱导小鼠产生良好的特异性体液和细胞免疫反应。结论含H5NA—HA的重组腺病毒疫苗可以诱导小鼠产生良好的免疫反应，为研制人禽流感疫苗打下基础。

Objective To construct adenovirus vector vaccine against H5N1 influenza virus and study on the immunogenicity. Methods In this study, we amplified hemagglutinin （HA） gene sequence of H5N1 influenza virus （A/Anhui/1/2005）, then constructed an adenovirus vector vaccine （Adv-HA）, followed by tests in BALB/c mice for the immunogenicity with the vaccine and immunization strategies. Results The recombinate Adv-HA vaccine could effectively induce both humoral and cellular immunity against human H5N1 influenza virus. Conclusion The Adv-HA vaccination against H5N1 influenza is a potential strategy and worthy of further investigation.