陈勍, 郭颖. 丝状病毒进入抑制剂的细胞水平评价体系的建立J. 药学学报, 2015,50(12): 1538-1544.
引用本文: 陈勍, 郭颖. 丝状病毒进入抑制剂的细胞水平评价体系的建立J. 药学学报, 2015,50(12): 1538-1544.
CHEN Qing, GUO Ying. Establishment of a cell-based filovirus entry inhibitor evaluation systemJ. Acta Pharmaceutica Sinica, 2015,50(12): 1538-1544.
Citation: CHEN Qing, GUO Ying. Establishment of a cell-based filovirus entry inhibitor evaluation systemJ. Acta Pharmaceutica Sinica, 2015,50(12): 1538-1544.

丝状病毒进入抑制剂的细胞水平评价体系的建立

Establishment of a cell-based filovirus entry inhibitor evaluation system

  • 摘要: 埃博拉病毒属于丝状病毒家族,可引起人类恶性传染病。本文旨在建立可用于高通量筛选评价丝状病毒进入抑制剂的细胞水平重组病毒模型。基于重组病毒技术,共建立了三株丝状重组病毒模型,分别是扎伊尔型埃博拉重组病毒、苏丹型埃博拉重组病毒和马尔堡重组病毒。该模型以丝状病毒表面唯一负责病毒进入的糖蛋白为病毒表面蛋白,包裹带有荧光素酶报告基因的HIV内核,组装为丝状重组病毒颗粒。当病毒感染细胞后,病毒感染程度可通过检测细胞中荧光素酶活性获得。此模型靶点明确,安全性好,当进行丝状病毒进入抑制剂筛选时,可通过与水泡性口膜炎重组病毒联用判断化合物的特异性。应用此模型对文献报道的丝状病毒进入抑制剂进行评价,数据与文献报道相符,模型构建成功。丝状重组病毒进入模型的建立对于抗丝状病毒药物的快速筛选和研发有重要意义。

     

    Abstract: Ebola virus, the cause of severe and fatal hemorrahagic fever in humans, belongs to filovirus family. This study was designed to establish a cell-based screening and evaluation system in the pharmacological study of antivirus compounds. Three reporter systems were established with recombinant pseudoviral luciferase of HIV core(pNL4-3.Luc.R-E-) packed with filovirus glycoprotein(EBOV-Zaire GP/HIV-luc, EBOV-Sudan GP/HIV-luc and Marburg GP/HIV-luc), which are required for virus entry of cells. The level of filovirus entry was determined by the expression of luciferase reporter gene in the infected cells. For screening of filovirus entry inhibitors, the vesicular stomatitis G packed pseudovirions(VSVG/HIV-luc) was used to determine the compound specificity. The results of known filovirus entry inhibitors demonstrated successful establishment of the new model systems, which would be useful in high throughput screening of anti-filovirus drugs in the future.

     

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