H5N1流感疫苗株在Vero细胞中的研究[英文]  

Development of prepared H5N1 influenza vaccine strain in Vero cells

作　　者：鹿文葆[1] 罗德炎[1] 杨鹏辉[1] 刘坤[1] 刑丽[1] 王希良[1] 刘秀梵[2] 

机构地区：[1]军事医学科学院微生物流行病学研究所病原微生物生物安全国家重点实验室,北京100071 [2]扬州大学农业部畜禽传染病学重点实验室,225009

出　　处：《免疫学杂志》2010年第4期281-284,289,共5页Immunological Journal

基　　金：supported by grants(2006AA02A204)from the National“863”great item~~

摘　　要：目的产生在Vero(非洲绿猴肾)细胞中能高适应生长的重配H5N1流感病毒疫苗株,并对其生物学特性进行初步测定。方法修饰A/Puerto Rico/8/34(H1N1)(PR8)的NS基因为Vero细胞适应型,并合成NS基因片段,构建质粒pHW2000-NS;通过RT-PCR方法,扩增疫苗株A/Anhui/01/2005(H5N1)的HA、NA基因片段,构建质粒pHW2000-HA、pHW2000-NA;以PR8的其它5个内部基因作为骨架,按照1+2+5模式8质粒共转染Vero细胞拯救流感病毒,并对拯救病毒的生物学特性进行检测。结果由Vero拯救出具有高适应生长特性的H5N1亚型人禽流感病毒疫苗株,并检测了重配疫苗株的生物学特性。结论成功从Vero细胞拯救了具有Vero细胞高适应性生长特性的H5N1亚型人禽流感病毒疫苗株,为应用Vero细胞大规模生产流感疫苗奠定了基础。The aim was to generate an attenuated and high-yielding H5N1 HPAI virus candidate vaccine from African green monkey kidney（Vero） cells by reverse genetics and to analyze its phenotype characters.The NS gene of A/Puerto Rico/8/34（H1N1） （PR8） was modified with the aim of Vero-adapting,and then synthesized to construct plasmid pHW2000;hemagglutinin（HA） and neuramindase（NA） genes of A/Anhui/01/2005（H5N1） amplified by RT-PCR were cloned into the plasmid pHW2000 for constructing pHW2000-HA and pHW2000-NA.The three plasmids above were cotransfected into Vero cell with other 5 plasmids from PR8 virus recovery system in order to generate the virus.The reassortant H5N1 influenza virus vaccine candidate strain contained the HA antigen phenotype of A/Anhui/01/2005（H5N1） was propagated in Vero cell in a highly-adapted manner.The growth trend of the reassortant virus in Vero cells was significantly improved in comparison to those of viruses rescued with PR8 recovery system.The reassortant H5N1 influenza virus with high adaptability could replicate in Vero cells.And the successfully recovery of H5N1 influenza virus vaccine strain paved the way for large scale produce of influenza vaccine in Vero cells which will benefit influenza epidemic preventing.

关 键 词：H5N1流感疫苗 反向遗传学 VERO细胞 

分 类 号：R392[医药卫生—免疫学]