Institute of Medical and Veterinary Science, Adelaide, SA, Australia.
J Virol Methods. 2009 May;157(2):161-7. doi: 10.1016/j.jviromet.2008.12.016. Epub 2009 Jan 17.
Four IgG(1kappa) monoclonal antibodies (mAbs) against Influenza A/Chicken/Vietnam/8/2004 (H5N1) virus are described. Three of these showed neutralizing activities against H5N1 strains from clades 1, 2 and 3 using a retroviral pseudotype or live virus microneutralization assay. In the pseudotype assay, the IC(90) neutralizing titre range was >1600-51,200, and with the microneutralization was 80> or =10,240. MAb 1C1 showed strong neutralizing activities in both assays. All four mAbs reacted specifically to the immunogen by immunohistochemical staining and to A/Hong Kong/483/1997 (H5N1) and A/Thailand/1(KAN-1)/2004 (H5N1)-infected MDCK cells by immunofluorescence. ELISA titrations of the mAbs showed specificity for H5N1 haemagglutinin (HA) and no cross-reactivity to 15 other Influenza A subtypes. Only mAbs 1C1 and the non-neutralizing 1F7 reacted with HA(1), the cleaved subunit of HA, by Western blot. These results suggest that the mAbs recognize distinct or overlapping epitopes and will be useful reagents for construction of specific rapid point-of-care assays or for therapeutic use.
描述了针对甲型流感/鸡/越南/8/2004（H5N1）病毒的四种 IgG(1kappa)单克隆抗体 (mAb)。其中三种 mAb 在使用逆转录病毒假型或活病毒微量中和测定法时对来自 1、2 和 3 类群的 H5N1 株具有中和活性。在假型测定法中，IC(90)中和效价范围为>1600-51,200，在微量中和测定法中为 80>或 =10,240。 mAb 1C1 在两种测定法中均显示出强大的中和活性。所有四种 mAb 均通过免疫组织化学染色特异性地针对免疫原，以及通过免疫荧光特异性地针对 A/香港/483/1997（H5N1）和 A/泰国/1(KAN-1)/2004（H5N1）感染的 MDCK 细胞进行反应。 mAb 的 ELISA 滴定显示出对 H5N1 血凝素 (HA) 的特异性，与 15 种其他甲型流感亚型没有交叉反应性。只有 mAb 1C1 和非中和性的 1F7 通过 Western blot 与 HA(1)（HA 的裂解亚单位）反应。这些结果表明，mAb 识别独特或重叠的表位，将成为用于构建特定即时护理点测定法或用于治疗用途的有用试剂。
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