Key Laboratory of Molecular Virology & Immunology, Institut Pasteur of Shanghai, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, 411 Hefei Road, Shanghai 200025, China.
J Virol Methods. 2012 Dec;186(1-2):193-7. doi: 10.1016/j.jviromet.2012.06.025. Epub 2012 Jul 4.
Enterovirus 71 (EV71) is a member of the Picornaviridae family and one of the main causative agents of hand, foot, and mouth disease (HFMD). Currently, EV71 infection is prevalent in the Asia-Pacific regions where it affects millions of children under the age of five, causing significant morbidity and mortality. No specific vaccine or antiviral drugs are available for EV71. The development of murine monoclonal antibodies (mAbs) with potent neutralization effects on EV71 is described. Mab-secreting hybridomas were generated from mice immunized with EV71 recombinant virus-like particles. Three IgG1 mAbs, D5, H7, and C4, capable of binding to and neutralizing EV71, were identified. In ELISA and Western blot assays, these mAbs reacted with recombinant VP1 protein, but not with VP0. They also detected cells infected with EV71 by immunofluorescent staining. In addition, these three mAbs had potent EV71 neutralization capacity, with 95% inhibitory concentrations of 0.3125, 0.3125, and 1.25 μg/ml for D5, H7, and C4, respectively. The presented data demonstrate that the anti-EV71 mAbs are not only promising candidates for development into humanized mAb for treatment but also useful reagents for development of diagnostic tests.
肠道病毒 71 型(EV71)属于小 RNA 病毒科肠道病毒属,是手足口病(HFMD)的主要病原体之一。目前,EV71 感染在亚太地区流行,每年有上百万 5 岁以下儿童感染,导致较高的发病率和死亡率。EV71 目前尚无特异性疫苗和抗病毒药物。本研究旨在开发对 EV71 具有高效中和作用的鼠源性单克隆抗体(mAbs)。我们用 EV71 重组病毒样颗粒免疫小鼠,制备分泌 mAbs 的杂交瘤细胞株。通过鉴定,获得了 3 株能够结合并中和 EV71 的 IgG1 型 mAbs,分别命名为 D5、H7 和 C4。ELISA 和 Western blot 结果表明,这些 mAbs 与重组 VP1 蛋白反应,而与 VP0 无反应。免疫荧光染色结果显示,这些 mAbs 可检测到感染 EV71 的细胞。此外,这 3 株 mAbs 对 EV71 均具有较强的中和活性,D5、H7 和 C4 的 95%抑制浓度(IC50)分别为 0.3125、0.3125 和 1.25 μg/ml。研究结果表明,这些抗-EV71 mAbs 不仅是开发治疗用人源化 mAb 的有前途的候选物,也是开发诊断检测试剂的有用试剂。