一株H5N2亚型禽流感病毒细胞适应株的培育及免疫原性研究  

Cultivation of a cell-adapted H5N2 subtype avian influenza virus strain and its immunogenicity analysis

作　　者：张凯[1] 高瑞娟[1] 黄武[1] 黄永兴[1] 吕嘉敏 罗开健[1] 

机构地区：[1]华南农业大学兽医学院,广东广州510642

出　　处：《中国兽医科学》2014年第2期129-133,共5页Chinese Veterinary Science

基　　金：公益性行业(农业)科研专项(201303044)

摘　　要：为获得可在MDCK细胞上高水平复制的H5N2亚型禽流感病毒(AIV)疫苗候选株,将1株H5N2亚型AIV在MDCK细胞上进行传代驯化,检测各代次病毒的血凝(HA)效价以及组织培养半数感染量(TCID50),选取细胞培养病毒液与鸡胚培养病毒液分别制备油乳剂灭活疫苗,免疫4周龄SPF鸡,于免疫后不同时间检测抗体效价,免疫后第21天测定疫苗的免疫保护情况。结果显示,病毒经过传代后逐步适应了MDCK细胞,其HA效价趋于稳定,最高时与鸡胚传代病毒(9lb)一致,TCID50接近亲本毒株,HA、NA基因序列与亲本毒株基本保持一致,免疫后第21天细胞毒免疫组的HI抗体平均效价(8.5lb)与鸡胚毒株免疫组的效价(9lb)基本相当,而且细胞毒免疫组能对H5N1亚型AIV毒株产生良好的免疫保护作用,抑制免疫鸡排毒。结果表明,该细胞适应株的培育为H5N2亚型AIV细胞疫苗的研制奠定了基础。To obtain a high-yield H5N2 subtype avian influenza virus（AIV） candidate vaccine strain in MDCK cells,a HSN2 subtype AIV strain was serially passaged in MDCK cells. HA titer and TCID50 of each passage were determined. The supernatant of virus culture and the chicken embryo allantoic virus cul-ture were prepared for the development of oil-emulsion inactivated vaccine,and 4-weeks-old SPF chickens were immunized. Then HI antibody titer was detected at different time points after immunization and the protection was evaluated on day 21 post-vaccination. In result,the virus gradually adapted to MDCK cells through serial passages. The cell-adapted virus shared similar biological properties with the parental strain, such as HA,TCID50 and genetic stability. The HI antibody titer had no significant difference between cell-adapted virus vaccine（8.51b）and chicken embryo vaccine（91b） on day 21 post-vaccination,and cell-adapted virus vaccine could induce desirable protection against the H5N1 strain. The results lay a solid foundation for the future development of HSN2 subtype AIV cell vaccine.

关 键 词：禽流感病毒 H5N2亚型 细胞疫苗 

分 类 号：S852.659.5[农业科学—基础兽医学]