Characterization of human single-chain antibodies against highly pathogenic avian influenza H5N1 viruses : mimotope and neutralizing activity
Characterization of human single-chain antibodies against highly pathogenic avian influenza H5N1 viruses : mimotope and neutralizing activity
复制标题
抗高致病性禽流感 H5N1 病毒的人单链抗体的表征：模拟表位和中和活性
DOI：
10.1093/jb/mvq084
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发表时间：
2010
影响因子：
2.7
通讯作者：
K.
中科院分区：
文献类型：
--
作者：
Yang;J.;Yoshida;R.;Kariya;Y.;Zhang;X.;Hashiguchi;S.;Nakashima;T.;Suda;Y.;Takada;A.;Ito;Y.;Sugimura;K.
目前迫切需要开发新的治疗靶点和策略来控制人类高致病性禽流感（HPAI）H5 N1病毒感染。中和重组人抗体将为H5 N1病毒感染的免疫治疗和疫苗开发关键模拟表位的确定提供重要的试剂。在这项研究中，我们已经确定了一个抗H5特异性的单链抗体克隆，3D 1从人的单链抗体展示噬菌体库。3D 1在流式细胞术中阻断H5-Fc与MDCK细胞的结合，并在微量中和测定中中和H5 N1亚型甲型流感病毒。利用噬菌体噬菌体展示肽库，Ph.D-12，确定模拟表位在H5的#128-131和#204-211，这是硅酸结合区。与该结果一致，3D 1结合由杆状病毒载体产生的重组糖结合结构域（#50G-#272E）。3D 1抗体采用种系基因VH 1 -23。由于该抗体是第一个在糖结合表位上明确定义的人源抗H5单链抗体，它允许我们研究该区域中的氨基酸取代对确定唾液酸α 2，6-半乳糖（SA α2，6 Gal）或唾液酸α 2，3-半乳糖（SA α2，3Gal）的结合特异性的影响，为有效的H5 N1大流行性疫苗的开发提供新的见解。
The development of new therapeutic targets and strategies to control highly pathogenic avian influenza (HPAI) H5N1 virus infection in humans is urgently needed. Neutralizing recombinant human antibodies would provide important agents for immunotherapy on human H5N1 virus infection and definition of the critical mimotope for vaccine development. In this study, we have characterized an anti-H5-specific scFv clone, 3D1 from the human-scFv-displaying phage library. 3D1 blocked the binding of H5-Fc to MDCK cells in flow cytometry and neutralized H5N1 subtype influenza A viruses in a microneutralization assay. Employing a peptide-displaying phage library, Ph.D-12, the mimotope was determined to be at #128-131 and #204-211 of H5, which are silic acid-binding regions. In consistency with this result, 3D1 binds the recombinant sugar-binding domain (#50G-#272E) produced by a baculovirus vector. The 3D1 antibody employs the germline gene VH1-23. As this antibody is the first human anti-H5 scFv clearly defined on the sugar-binding epitope, it allows us to investigate the influence of amino acid substitutions in this region on the determination of the binding specificity to either sialic acid α2,6-galactose (SA α2,6Gal) or sialic acid α2,3-galactose (SA α2,3Gal) providing new insight for the development of effective H5N1 pandemic vaccines.