IR@PKUHSC  > 北京大学基础医学院  > 病原生物学系
学科主题基础医学
A 3C(pro)-dependent bioluminescence imaging assay for in vivo evaluation of anti-enterovirus 71 agents
Guo, Zhiwei1; Zhong, Xiaoyan1; Lin, Lexun1; Wu, Shuo1; Wang, Tianying1; Chen, Yang1; Zhai, Xia1; Wang, Yan1; Wu, Heng1; Tong, Lei1; Han, Yelu1; Pan, Bo1; Peng, Yihong3; Si, Xiaoning1; Zhang, Fengmin1; Zhao, Wenran2; Zhong, Zhaohua1
关键词Enterovirus 71 Split luciferase 3C protease In vivo imaging
刊名ANTIVIRAL RESEARCH
2014
DOI10.1016/j.antiviral.2013.11.002
101期:1页:82-92
收录类别SCI
文章类型Article
WOS标题词Science & Technology
类目[WOS]Pharmacology & Pharmacy ; Virology
资助者Natural Science Foundation of China (NSFC) ; Natural Science Foundation of China (NSFC)
研究领域[WOS]Pharmacology & Pharmacy ; Virology
关键词[WOS]COXSACKIEVIRUS B3 REPLICATION ; MOUTH-DISEASE ; ANTIVIRAL ACTIVITY ; KINASE INHIBITORS ; INFECTION MODEL ; CLINICAL-TRIAL ; DOUBLE-BLIND ; HAND ; FOOT ; CHILDREN
英文摘要

Enterovirus 71 (EV71), a member of Picornaviridae, is one of the major pathogens of human hand, foot and mouth disease. EV71 mainly infects children and causes severe neurological complications and even death. The pathogenesis of EV71 infection is largely unknown, and no clinically approved vaccine or effective treatment is available to date. Here we described a novel bioluminescence imaging approach for EV71 detection. In this approach, a plasmid-based reporter was constructed to express the fusion protein AmN(Q/G)BC, a split firefly luciferase mutant, which can be specifically cleaved by EV71 protease 3C(pro). Upon cleavage, the splitting fusion protein restores luciferase activity. Our test confirmed that AmN(Q/G)BC was specifically cleaved by 3C(pro) and EV71 and restored the luciferase activity to a degree that corresponds to the 3C(pro) and virus doses in cells and mice. The anti-EV71 effect of GW5074 and U0126, two mitogen-activated protein kinase (MAPK) inhibitors, was evaluated using this approach to validate its application of screening anti-EV71 agents. We found that the AmN(Q/G)BC reporter efficiently monitored the inhibitory effect of GW5074 and U0126 on EV71 infection under in vitro and in vivo conditions. The data from AmN(Q/G)BC reporter were consistent with Western blotting and histopathology examination. Taken together, this real-time imaging approach can quantitatively monitor the efficacy of anti-EV71 agents and is valuable for anti-EV71 drug screening and evaluation, especially, under in vivo conditions. (C) 2013 Elsevier B.V. All rights reserved.

语种英语
所属项目编号81271825 ; 31270198 ; 81101234 ; 81101235
资助者Natural Science Foundation of China (NSFC) ; Natural Science Foundation of China (NSFC)
WOS记录号WOS:000329772700011
Citation statistics
Cited Times:4[WOS]   [WOS Record]     [Related Records in WOS]
文献类型期刊论文
版本出版稿
条目标识符http://ir.bjmu.edu.cn/handle/400002259/63545
Collection北京大学基础医学院_病原生物学系
作者单位1.Harbin Med Univ, Dept Microbiol, Harbin 150081, Peoples R China
2.Harbin Med Univ, Dept Cell Biol, Harbin 150081, Peoples R China
3.Peking Univ, Hlth Sci Ctr, Dept Microbiol, Beijing 100191, Peoples R China
Recommended Citation
GB/T 7714
Guo, Zhiwei,Zhong, Xiaoyan,Lin, Lexun,et al. A 3C(pro)-dependent bioluminescence imaging assay for in vivo evaluation of anti-enterovirus 71 agents[J]. ANTIVIRAL RESEARCH,2014,101(1):82-92.
APA Guo, Zhiwei.,Zhong, Xiaoyan.,Lin, Lexun.,Wu, Shuo.,Wang, Tianying.,...&Zhong, Zhaohua.(2014).A 3C(pro)-dependent bioluminescence imaging assay for in vivo evaluation of anti-enterovirus 71 agents.ANTIVIRAL RESEARCH,101(1),82-92.
MLA Guo, Zhiwei,et al."A 3C(pro)-dependent bioluminescence imaging assay for in vivo evaluation of anti-enterovirus 71 agents".ANTIVIRAL RESEARCH 101.1(2014):82-92.
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