具有HI活性的H1亚型流感病毒特异性单克隆抗体的制备与应用  

Development monoclonal antibodies against the H1 subtype swine influenza virus

作　　者：李印[1] 邓国华[1] 王桂芹[1] 杨焕良[1] 乔传玲[1] 陈化兰[1] 

机构地区：[1]中国农业科学院哈尔滨兽医研究所农业部动物流感重点开放实验室/兽医生物技术国家重点实验室,黑龙江哈尔滨150001

出　　处：《中国预防兽医学报》2010年第4期307-311,共5页Chinese Journal of Preventive Veterinary Medicine

基　　金：国家十五重点科技攻关课题(2004BA519A054)

摘　　要：为了制备具有HI活性的HI亚型流感病毒特异性单克隆抗体(MAb),本研究以H1N1亚型猪流感病毒(SIV)株A/Swine/Guangdong/718/01(H1N1)为免疫原,免疫BALB/c小鼠,经常规细胞融合后,血凝抑制(HI)方法进行检测,融合细胞经稀释克隆纯化后,获得11株能稳定分泌抗血凝素特异性HI MAb的杂交瘤细胞株。鉴定表明,所获MAb与其他具有血凝活性的病毒以及其他14个HA亚型的流感病毒均不具有HI交叉反应,表明这11株MAb均具有良好的流感病毒亚型特异性。其中A6F、2BBF和2BB与其他H1亚型流感病毒分离株的HI试验证实我国不同地区分离株之间的抗原性存在一定差异。11株MAb对H1SIV抗原的HI试验结果显示其HI效价有明显差异。叠加实验表明这些MAb分别识别HA抗原的不同表位。间接免疫荧光试验表明,2BBF、8HB、1DH、7FC和2BB均可与2009年流行H1N1病毒A/California/04/2009HA抗原发生特异性反应。这些MAb特异性的研制为H1亚型流感病毒的疫情病原学快速诊断以及病毒抗原性变异的相关研究提供了物质基础。Eleven monoclonal antibodies（MAbs） against H1 subtype swine influenza virus（SIV） were developed by fusing between sp2/0 myeloma cells and spleen cells of BALB/c mice immunized with the H1 subtype SIV. These MAbs were highly specific to H1 subtype SIV,and did not react with AIV H2 to H15 subtype AIV and others viruses in the hemagglutin inhibition test（HI） . Furthermore,each MAb was directed against different epitope on HA,and exhibited different HI titer against H1 subtype SIV. The MAb A6F,2BBF and 2BB were shown to have varied HI capability against several H1 subtype influenza viruses,indicating that these viruse isolates may have distinct diversity in hemagglutin. The MAb 2BBF,8HB,1DH,7FC and 2BB were shown to react with the HA antigen of A/California/04/2009A（H1N1） in indirect-immunofluorescence assay. These results indicated that the MAbs could be useful in H1 subtype influenza virus surveillance and for the study of antigenic variation of HI viruses.

关 键 词：H1亚型猪流感 血凝抑制 单克隆抗体 间接免疫荧光 

分 类 号：S852.65[农业科学—基础兽医学]