从cDNA感染性克隆产生冷适应重配A型人流感病毒及免疫原性的初步研究  

Generation of a cold-adapted reassortant influenza A virus from cDNA and research on its immunogenicity

作　　者：叶艺[1] 杨鹏辉[2] 罗德炎[2] 吴朔[1] 周朋[1] 刘秀梵[3] 王希良[2] 于三科[1] 

机构地区：[1]西北农林科技大学动物医学院,陕西杨陵712100 [2]军事医学科学院微生物流行病研究所病原微生物生物安全国家重点实验室,北京100071 [3]扬州大学农业部畜禽传染病学重点开放实验室,江苏扬州225009

出　　处：《免疫学杂志》2008年第4期373-377,共5页Immunological Journal

基　　金：国家科技攻关资助项目(2004BA519A70)

摘　　要：目的从cDNA感染性克隆产生冷适应重配A型人流感病毒株,并对其生物学特性和免疫原性进行初步测定。方法采用RT-PCR技术,分别扩增2006至2007年疫苗株A/New Caledonia/20/99的HA、NA全长基因片段,构建转录表达双向载体pMDV-A-HA、pMDV-A-NA,将这2个质粒与冷适应拯救系统的6个质粒共转染COS-1细胞,并对拯救的重配病毒的生物学特性和免疫原性进行初步鉴定。结果拯救出具有冷适应表型和温度敏感型的A型人流感弱毒株,初步测定了重配病毒的生物学特性和免疫原性。结论成功拯救具有冷适应和温度敏感表型的A型人流感弱毒株,并初步测定其免疫原性,为我国弱毒疫苗株的拯救和弱毒疫苗的发展奠定了基础。Objective To generate the cold-adapted reassortant influenza A virus by reverse genetics from cDNAs and to analyze its phenotype characters and immunogenicity. Methods The hemagglutinin （HA） and neuramindase （NA） genes of A/New Caledonia/20/99 strain were amplified by RT-PCR, and then cloned into the plasmid pAD3000. The constructed bidirectional expression vectors, pMDV-A-HA and pMDV-A-NA, were transfected into the COS-1 cell together with other 6 plasmids from cold-adapted virus recovery system. The phenotype characters and immunogenicity of the reassortant influenza A vires were analyzed. Results The reassortant influenza A vires was recovered with cold-adapted phenotype and temperature sensitivity. And its bionomics and immunogenicity was analyzed. Conclusion The reassortant influenza A vires with cold-adapted phenotype and temperature sensitivity is recovered successfully and its immunogenicity is determined, which lays the foundation for recovering the human influenza attenuated strain and developing the influenza attenuated vaccine.

关 键 词：反向遗传学 冷适应 弱毒疫苗株 重配病毒 免疫原性 

分 类 号：R373[医药卫生—病原生物学]