不同亚型禽流感病毒红细胞凝集特性分析  

Analysis of erythrocyte agglutination characteristics of different subtypes of avian influenza virus

作　　者：李梓[1] 刘佳 李希妍[1] 杨磊[1] 陈志肖 吴宇超 王大燕[1] 刘丽琦[1] Li Zi;Liu Jia;Li Xiyan;Yang Lei;Chen Zhixiao;Wu Yuchao;Wang Dayan;Liu Liqi(National Influenza Center,National Institute for Viral Disease Control and Prevention,Chinese Center for Disease Control and Prevention,Beijing 102206 China)

机构地区：[1]中国疾病预防控制中心病毒病预防控制所国家流感中心,北京102206

出　　处：《中华实验和临床病毒学杂志》2021年第1期68-74,共7页Chinese Journal of Experimental and Clinical Virology

基　　金：国家科技重大专项"艾滋病和病毒性肝炎等重大传染病防治"(2018ZX10711-001)。

摘　　要：目的了解不同亚型禽流感病毒与不同动物来源红细胞的凝集特性,为流感环境样本监测工作选择更适宜的检测用红细胞。方法选取2009-2016年我国禽流感环境监测中分离到的不同亚型的禽流感毒株,采用红细胞凝集试验,选用5种动物红细胞(鸡、火鸡、豚鼠、马和绵羊)进行检测;应用流式细胞仪检测不同动物红细胞表面唾液酸受体的表达及类型,通过氨基酸序列分析病毒血凝素蛋白受体结合区(receptor binding domain,RBD)的特征。结果本研究共检测了14种亚型的28株禽流感病毒。结果显示所有毒株均能与火鸡和豚鼠红细胞发生凝集,其余红细胞均出现不能与某些毒株产生凝集的现象;其中1株H9N2(A/环境/安徽/43762/2015)毒株与鸡红细胞不产生凝集反应;1株H1N1(A/环境/山东/76972/2014)和2株H9N2(A/环境/重庆/79449/2014和A/环境/安徽/43762/2015)毒株与马红细胞不产生凝集反应;2株H9N2(A/环境/重庆/79449/2014和A/环境/安徽/43762/2015)和2株H13N8(A/环境/青海湖/166/2012和A/环境/青海湖/13/2012)毒株与绵羊红细胞不产生凝集反应。流式检测结果显示在火鸡、鸡和豚鼠红细胞表面可检测到两种类型唾液酸受体α2,3和α2,6,但两种受体表达比例不同;马和绵羊红细胞表面唾液酸受体只检测到表达α2,3。序列分析提示病毒RBD关键区域的氨基酸替换可能是影响病毒红细胞结合特性的重要因素。结论在禽流感病毒检测中,火鸡和豚鼠红细胞在红细胞凝集试验中敏感性最好。不同来源的红细胞其受体表达及类型会显著影响不同亚型禽流感病毒的凝集反应,同时RBD关键区域的氨基酸变化也会影响病毒与不同红细胞的结合。Objective To understand the agglutination characteristics of different subtypes of avian influenza viruses,we selected erythrocytes from different sources to find suitable erythrocytes for influenza environmental sample detection.Methods Different subtypes of avian influenza viruses,which were isolated from environmental sample between 2009 and 2016 in China,were selected to do hemagglutination assay using 5 animal erythrocytes(chicken,turkey,guinea pig,horse,and sheep).Flow cytometry was used to detect expression level and type of sialic acid receptors of different erythrocytes,and the characteristics of the receptor binding domain(RBD)of the viral hemagglutinin protein were analyzed by amino acid sequence.Results In this study,a total of 28 strains of avian influenza virus including 14 subtypes were detected.The result showed that all viruses could agglutinate with turkey and guinea pig erythrocytes and the rest three erythrocytes were unable to produce agglutination with some viruses;among them,one H9N2 virus(A/environment/Anhui/43762/2015)did not agglutinate with chicken erythrocytes,one H1N1 virus(A/environment/Shandong/76972/2014)and two H9N2 viruses(A/environment/Chongqing/79449/2014 and A/environment/Anhui/43762/2015)did not agglutinate with horse erythrocytes,two viruses of H9N2(A/environment/Chongqing/79449/2014 and A/environment/Anhui/43762/2015)and two viruses of H13N8(A/environment/Qinghai Lake/166/2012 and A/environment/Qinghai Lake/13/2012)did not agglutinate with sheep erythrocytes.The result of flow cytometry showed that two sialic acid receptors,α-2,3 andα-2,6,were detected on the surface of erythrocytes of turkey,chicken and guinea pig,but the expression ratios of the two receptors were different.Only the expression ofα-2,3 sialic acid receptors was detected in horse and sheep erythrocytes.Sequence analysis suggested that amino acid substitution in key regions of viral hemagglutinin protein RBD may be an important factor affecting the binding properties of different erythrocytes.Conclusions

关 键 词：禽流感病毒 红细胞 受体 受体结合区 

分 类 号：S852.65[农业科学—基础兽医学]