流感病毒H3N2小鼠适应株的建立及分子机制探究  

Establishment of a mouse-adapted seasonal influenza A H3N2 virus strain and preliminary inquiry its molecular mechanism

作　　者：胡云光[1] 徐娟 胡雅洁[1] 宋杰[1] 乞素冬[1] 秦晓峰 刘龙丁[1] 

机构地区：[1]中国医学科学院北京协和医学院医学生物学研究所 [2]中国医学科学院苏州系统医学研究所

出　　处：《中华实验和临床病毒学杂志》2015年第3期203-206,共4页Chinese Journal of Experimental and Clinical Virology

基　　金：国家自然科学基金面上项目（81373142）;云南省生物重大科技专项（2012ZA009）

摘　　要：目的 建立季节性甲型流感病毒influenza A virus[（A/Hong Kong/1968 （H3N2）]毒株的鼠肺适应株,并对其适应过程的分子机理进行初步探究.方法 以滴鼻的方式使小鼠感染H3N2流感病毒,并在小鼠肺部进行连续传代,通过对小鼠的临床观察、病毒载量及病毒滴度测定、肺部病理改变、细胞因子检测等方面的观察和检测,来获得季节性流感病毒influenza A virus[A/Hong Kong/1968（H3N2）]毒株的鼠肺适应株.结果 人季节性甲型流感病毒influenza A virus[A/Hong Kong/1968 （H3N2）]毒株,经过小鼠肺中连续7次传代以后,病毒毒力逐渐增强,鼠肺中病毒载量提高了4个数量级,病毒滴度上升了2.5lgCCID50,对流感病毒HA、NA两个节段的测序,发现这两个节段上均有一个有义突变.结论 在自然状态下并不感染小鼠的野生季节性流感病毒H3N2毒株可以通过在小鼠肺部中连续传代,得到毒力较高的鼠肺适应株,HA节段中248位的Ile突变为Val,NA节段中444位的Val突变为Ile,这两个突变位点可能与流感病毒毒力相关。Objective To establish adapted strain of seasonal influenza virus influenza A virus [A/ Hong Kong/1968 （H3N2）] in mice and explore the molecular mechanism of adaption preliminary.Methods The mice model was induced by nose dropping with wild type H3N2 influenza virus,by continuous passage in the lung of mice and detection of the clinical manifestation,viral load,virus titer,pathology,cytokines,to get the adapted strain of seasonal influenza A virus [A/Hong Kong/1968 （H3N2）].Results After 7 times continuous passage in the lung of mice,virulence was increased,viral load and virus titer were increased either.Genome sequencing and alignment indicated that the HA and NA gene was mutated.Conclusion The influenza virus H3N2 of mice lung adaption can be acquired from wild type seasonal H3N2 by continuous passage in the lung tissues of mice.Mutation of Ile to Val at residue 248 of HA and Val to Ile at residue 444of NA protein may be the key virulence determinant.

关 键 词：流感病毒A型 小鼠 近交BALBC 病毒培养 

分 类 号：R373[医药卫生—病原生物学]