Enterovirus type 71-immunized chicken egg yolk immunoglobulin has cross antiviral activity against coxsackievirus A16 in vitro
Enterovirus type 71-immunized chicken egg yolk immunoglobulin has cross antiviral activity against coxsackievirus A16 in vitro
复制标题
肠道病毒71型免疫鸡蛋黄免疫球蛋白体外对柯萨奇病毒A16具有交叉抗病毒活性
DOI：
10.3892/etm.2019.7529
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发表时间：
2019-07-01
影响因子：
2.7
通讯作者：
Wei, Jingchen
中科院分区：
文献类型：
--
作者：
Gao, Enyi;Wu, Shuwen;Wei, Jingchen
为探索肠道病毒引起的手足口病（HFMD）的交叉被动免疫治疗方法，研究了肠道病毒71型（EV 71）和柯萨奇病毒A16型（CVA 16）中和抗体的体外交叉抗病毒活性。用EV 71抗原免疫白色来亨鸡，从鸡蛋黄中制备特异性分离免疫球蛋白（IgY）。纯化后的IgY经SDS-PAGE、ELISA、Western blotting和双向免疫琼脂扩散试验鉴定。通过体外对横纹肌肉瘤（RD）细胞的细胞病变效应，测定IgY的抗病毒活性和剂量反应。结果表明，与阴性对照组相比，免疫后第7天IgY水平升高，第7周达到峰值，并持续4周。Western blotting和双向免疫琼脂扩散试验结果表明，该IgY通过靶向EV 71和CVA 16的包膜蛋白VP 0、VP 1和VP 3，在EV 71和CVA 16中具有交叉结合特性。中和试验结果表明，IgY对EV 71和CVA 16毒株在RD细胞中的感染性具有剂量依赖性的交叉阻断作用。综上所述，这些发现表明IgY在体外对EV 71和CVA 16具有交叉抗病毒活性，并且可能被开发为EV 71和CVA 16诱导的HFMD的被动免疫治疗。
To exploit a cross passive immunotherapy for enterovirus-induced hand-foot-and-mouth disease (HFMD), the cross antiviral activity of a neutralizing antibody against enterovirus 71 (EV71) and coxsackievirus A16 (CVA16) was investigated in vitro. White Leghorn specific-pathogen-free chickens were immunized with EV71 antigens and a specific isolated immunoglobulin (IgY) was prepared from the chicken egg yolk. IgY was further purified and characterized by SDS-PAGE, ELISA, western blotting and bidirectional immune agar diffusion testing. The antiviral activity and dose-response of the IgY were determined by assessing the cytopathic effect in rhabdomyosarcoma (RD) cells in vitro. It was indicated that the levels of IgY were increased at day 7, peaked at week 7 and were maintained at a higher level for 4 weeks following immunization when compared with the negative control. The results of western blotting and bidirectional immune agar diffusion testing revealed that the IgY had cross-binding properties in EV71 and CVA16 strains through targeting the envelope proteins (VP0, VP1 and VP3) of EV71 and CVA16. Neutralization assay results indicated that the infectivity of EV71 and CVA16 strains in RD cells was cross-blocked by IgY in a dose-dependent manner. To conclude, these findings indicate that IgY has cross antiviral activity against EV71 and CVA16 in vitro, and could potentially be developed as a passive immunotherapy for EV71- and CVA16-induced HFMD.