CAJ | 학술논문

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H1 SⅣ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/04/2009 HA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.
위료제비구유HI활성적HI아형류감병독특이성단극륭항체(MAb),본연구이H1N1아형저류감병독(SIV)주A/Swine/Guangdong/718/01(H1N1)위면역원,면역BALB/c소서,경상규세포융합후,혈응억제(HI)방법진행검측,융합세포경희석극륭순화후,획득11주능은정분비항혈응소특이성HI MAb적잡교류세포주.감정표명,소획MAb여기타구유혈응활성적병독이급기타14개HA아형적류감병독균불구유HI교차반응,표명저11주MAb균구유량호적류감병독아형특이성.기중A6F、2BBF화2BB여기타H1아형류감병독분리주적HI시험증실아국불동지구분리주지간적항원성존재일정차이.11주MAb대H1 SⅣ항원적HI시험결과현시기HI효개유명현차이.첩가실험표명저사MAb분별식별HA항원적불동표위.간접면역형광시험표명,2BBF、8HB、1DH、7FC화2BB균가여2009년류행H1N1병독A/California/04/2009 HA항원발생특이성반응.저사MAb특이성적연제위H1아형류감병독적역정병원학쾌속진단이급병독항원성변이적상관연구제공료물질기출.
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 AJV 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.