Construction of coxsackievirus B5 viruses with luciferase reporters and their applications <i>in vitro</i> and <i>in vivo</i>

Shangrui Guo; Meng Xun; Tingting Fan; Xinyu Li; Haoyan Yao; Xiaozhen Li; Bo Wu; Hang Yang; Chaofeng Ma; Hongliang Wang

doi:10.1016/j.virs.2023.05.010

August 2023

. doi: 10.1016/j.virs.2023.05.010

Citation: Shangrui Guo, Meng Xun, Tingting Fan, Xinyu Li, Haoyan Yao, Xiaozhen Li, Bo Wu, Hang Yang, Chaofeng Ma, Hongliang Wang. Construction of coxsackievirus B5 viruses with luciferase reporters and their applications in vitro and in vivo .VIROLOGICA SINICA, 2023, 38(4) : 549-558. http://dx.doi.org/10.1016/j.virs.2023.05.010

带有荧光素酶报告基因的柯萨奇B5病毒构建及其体内和体外应用

郭尚瑞 ^a ,
寻萌 ^a ,
范婷婷 ^a ,
黎欣宇 ^a ,
姚昊妍 ^b ,
李晓珍 ^a ,
吴博 ^a ,
杨行 ^a ,
马超峰 ^c ,
王洪亮 ^a,,

a. 西安交通大学医学部病原生物学与免疫学系;
b. 西安交通大学第一附属医院妇产科;
c. 西安市疾病预防控制中心病毒检验科

通讯作者： 王洪亮, hongliangwang@xjtu.edu.cn
收稿日期： 2023-02-01
录用日期： 2023-05-11

摘要

柯萨奇病毒属于小核糖核酸病毒科，是引起婴幼儿手足口病(HFMD)的主要病原体之一，可能导致患者出现严重并发症甚至死亡。这种病毒的发病机制尚未完全阐明，也没有被批准的疫苗或抗病毒药物。本研究构建了柯萨奇病毒B5型病毒的全长感染性cDNA克隆，重组病毒表现出与亲本病毒相似的生长动力学和引起细胞病变的能力。然后在此基础上将荧光素酶报告基因嵌入生成全长或者亚基因组复制子(SGR)报告子病毒。全长报告病毒适合用于高通量抗病毒药物筛选，而SGR则是研究病毒与宿主相互作用的有用工具。更重要的是，我们构建的全长报告病毒也被证明可以感染哺乳小鼠模型，并且可以使用体内成像系统检测到报告基因，从而为体内追踪病毒提供了有力的工具。总之，我们成功构建了柯萨奇病毒B5报告病毒，并为体外和体内研究病毒-宿主相互作用以及高通量筛选(HTS)鉴定新的抗病毒药物提供了独特的工具。
- 柯萨奇病毒
- , 荧光素酶
- , 高通量筛选
- , 小鼠模型

Construction of coxsackievirus B5 viruses with luciferase reporters and their applications in vitro and in vivo

Shangrui Guo ^a ,
Meng Xun ^a ,
Tingting Fan ^a ,
Xinyu Li ^a ,
Haoyan Yao ^b ,
Xiaozhen Li ^a ,
Bo Wu ^a ,
Hang Yang ^a ,
Chaofeng Ma ^c ,
Hongliang Wang ^a,,

a. Department of Pathogen Biology and Immunology, Xi’an Jiaotong University Health Science Center, Xi’an, 710061, China;
b. Department of Gynecology and Obstetrics, The First Affiliated Hospital of Xi'an Jiaotong University, Xi’an, 710061, China;
c. Department of Viral Diseases Laboratory, Xi’an Center for Disease Control and Prevention, Xi’an, 710061, China

Corresponding author: Hongliang Wang, hongliangwang@xjtu.edu.cn
Received Date: 01 February 2023
Accepted Date: 11 May 2023

Abstract

Coxsackievirus belongs to the Picornaviridae family and is one of the major pathogens that cause hand, foot and mouth disease (HFMD) in infants and children with potential serious complications and even deaths. The pathogenesis of this virus is not fully elucidated and no vaccine or antiviral drug has been approved. In this study, a full-length infectious cDNA clone of coxsackievirus B5 virus was assembled and the recombinant virus displayed similar growth kinetics and ability to cause cytopathic effects as the parental virus. Luciferase reporter was then incorporated to generate both full-length and subgenomic replicon (SGR) reporter viruses. The full-length reporter virus is suitable for high-throughput antiviral screening, while the SGR is a useful tool to study viral-host interactions. More importantly, the full-length reporter virus has also been shown to infect the suckling mouse model and the reporter gene could be detected using an in vivo imaging system, thus providing a powerful tool to track viruses in vivo. In summary, we have generated coxsackievirus B5 reporter viruses and provided unique tools for studying virus-host interactions in vitro and in vivo as well as for high-throughput screenings (HTS) to identify novel antivirals.
- Coxsackievirus
- , Luciferase
- , High-throughput screening
- , Mouse model

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Electronic Supplementary Material

10.1016j.virs.2023.05.010-ESM.docx