A noninfectious pseudovirus system for an emerging orthoflavivirus

Yanfan Shi; Yu He; Xiaoli Wang; Zhen Wu; Tao Wang; Mingshu Wang; Renyong Jia; Dekang Zhu; Mafeng Liu; Xinxin Zhao; Qiao Yang; Ying Wu; Shaqiu Zhang; Juan Huang; Xumin Ou; Di Sun; Anchun Cheng; Shun Chen

doi:10.1016/j.virs.2025.04.006

June 2025

. doi: 10.1016/j.virs.2025.04.006

Citation: Yanfan Shi, Yu He, Xiaoli Wang, Zhen Wu, Tao Wang, Mingshu Wang, Renyong Jia, Dekang Zhu, Mafeng Liu, Xinxin Zhao, Qiao Yang, Ying Wu, Shaqiu Zhang, Juan Huang, Xumin Ou, Di Sun, Anchun Cheng, Shun Chen. A noninfectious pseudovirus system for an emerging orthoflavivirus .VIROLOGICA SINICA, 2025, 40(3) : 495-498. http://dx.doi.org/10.1016/j.virs.2025.04.006

一种新兴正黄病毒的非感染性假病毒系统构建

石炎凡 ^a ,
贺煜 ^a,b,c,d,e ,
王小利 ^a ,
吴震 ^a,b,c,d,e ,
王涛 ^a,b,c ,
汪铭书 ^a,b,c,e ,
贾仁勇 ^a,b,c,e ,
朱德康 ^a,b,c,e ,
刘马峰 ^a,b,c,e ,
赵新新 ^a,b,c,e ,
杨乔 ^a,b,c,e ,
吴英 ^a,b,c,e ,
张沙秋 ^a,b,c,e ,
黄娟 ^a,b,c,e ,
欧旭敏 ^a,b,c,d,e ,
孙迪 ^a,b,c,e ,
程安春 ^a,b,c,e,, ,
陈舜 ^a,b,c,d,e,,

a 四川农业大学动物医学院;
b 四川农业大学动物医学院禽病中心;
c 动物疫病与人类健康四川省重点实验室;
d 农业生物信息教育部重点实验室;
e 西南动物疫病防控技术教育部工程研究中心

通讯作者： 程安春, chenganchun@vip.163.com; 陈舜, shunchen@sicau.edu.cn
收稿日期： 2025-01-04
录用日期： 2025-04-17

摘要

乌苏图病毒（USUV）是一种新兴病原体，与西尼罗河病毒，日本脑炎病毒（JEV）同属于黄病毒科正黄病毒属。虽然USUV目前被归类为生物安全2级（BSL-2）病原体，但出于安全考虑，活病毒实验通常在生物安全3级设施中进行。目前USUV仅建立了具有传染性的克隆体。为了满足生物安全要求，同时建立研究其病毒复制等分子特征的实验体系，本研究构建了稳定表达USUV-NS1蛋白的BHK-21-NS1细胞系，并基于代表性毒株Turdus merula Netherlands 2016（TMN）成功建立了ΔNS1假病毒系统。该系统可支持TMN-ΔNS1病毒的高效复制，且在传代过程中不产生复制能力强的重组病毒。进一步研究表明，TMN-ΔNS1可以被异源正黄病毒（Kunjin virus 与Tembusu virus）的NS1反式补充，并支持复制。该系统完整模拟了病毒侵入、组装和释放等关键生命过程，为深入解析病毒-宿主互作机制及加速USUV感染治疗方案研发提供了重要技术平台。
- 假病毒
- , 正黄病毒
- , 乌苏图病毒
- , 稳转细胞系
- , NS1蛋白
- , 反式互补

A noninfectious pseudovirus system for an emerging orthoflavivirus

Yanfan Shi ^a ,
Yu He ^a,b,c,d,e ,
Xiaoli Wang ^a ,
Zhen Wu ^a,b,c,d,e ,
Tao Wang ^a,b,c ,
Mingshu Wang ^a,b,c,e ,
Renyong Jia ^a,b,c,e ,
Dekang Zhu ^a,b,c,e ,
Mafeng Liu ^a,b,c,e ,
Xinxin Zhao ^a,b,c,e ,
Qiao Yang ^a,b,c,e ,
Ying Wu ^a,b,c,e ,
Shaqiu Zhang ^a,b,c,e ,
Juan Huang ^a,b,c,e ,
Xumin Ou ^a,b,c,d,e ,
Di Sun ^a,b,c,e ,
Anchun Cheng ^a,b,c,e,, ,
Shun Chen ^a,b,c,d,e,,

a. Institute of Veterinary Medicine and Immunology, Sichuan Agricultural University, Chengdu, 611130, China;
b. Research Center of Avian Disease, College of Veterinary Medicine, Sichuan Agricultural University, Chengdu, 611130, China;
c. Key Laboratory of Animal Disease and Human Health of Sichuan Province, Sichuan Agricultural University, Chengdu, 611130, China;
d. Key Laboratory of Agricultural Bioinformatics, Ministry of Education, Chengdu, 611130, China;
e. Engineering Research Center of Southwest Animal Disease Prevention and Control Technology for Ministry of Education, Sichuan Agricultural University, Chengdu, 611130, China

Corresponding author: Anchun Cheng, chenganchun@vip.163.com Shun Chen, shunchen@sicau.edu.cn
Received Date: 04 January 2025
Accepted Date: 17 April 2025

Abstract

Highlights
1. A trans-complementation system for Usutu virus (USUV) recapitulates authentic viral replication.
2. High yield of TMN-ΔNS1 virus was obtained with no recombination into replication competent viruses.
3. TMN-ΔNS1 can be trans-complemented by heterologous orthoflavivirus NS1.

References
1. Bates, T.A., Chuong, C., Hawks, S.A., Rai, P., Duggal, N.K.,Weger-Lucarelli, J., 2021. Development and characterization of infectious clones of two strains of Usutu virus. Virology, 554, 28-36.
2. Blazquez, A.B., Escribano-Romero, E., Merino-Ramos, T., Saiz, J.C.,Martin-Acebes, M.A., 2013. Infection with Usutu virus induces an autophagic response in mammalian cells. PLoS Negl Trop Dis, 7, e2509.
3. Cvjetkovic, I.H., Petrovic, T., Petric, D., Milosevic, U., Radovanov, J., Kovacevic, G., Galovic, A.J., Patic, A., Nikolic, N.,Cvjetkovic, D., 2017. Usutu virus: an emerging flavivirus in Europe. Archives of Veterinary Medicine, 10, 25-35.
4. Golding, M., Seechurn, N., Baylis, M.,Johnson, N., 2023. JMM Profile: Usutu virus. J Med Microbiol, 72, 2.
5. Li, N., Zhang, Y.N., Deng, C.L., Shi, P.Y., Yuan, Z.M.,Zhang, B., 2019. Replication-Defective West Nile Virus with NS1 Deletion as a New Vaccine Platform for Flavivirus. J Virol, 93, e00720-00719.
6. Lindenbach, B.D.,Rice, C.M., 1997. trans-Complementation of yellow fever virus NS1 reveals a role in early RNA replication. J Virol, 71, 9608-9617.
7. Roesch, F., Fajardo, A., Moratorio, G.,Vignuzzi, M., 2019. Usutu Virus: An Arbovirus on the Rise. Viruses, 11, 640.
8. Van Leur, S.W., Heunis, T., Munnur, D.,Sanyal, S., 2021. Pathogenesis and virulence of flavivirus infections. Virulence, 12, 2814-2838.
9. Youn, S., Ambrose, R.L., Mackenzie, J.M.,Diamond, M.S., 2013. Non-structural protein-1 is required for West Nile virus replication complex formation and viral RNA synthesis. Virol J, 10, 339.
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通讯作者: 陈斌, bchen63@163.com

1.
沈阳化工大学材料科学与工程学院沈阳 110142

A noninfectious pseudovirus system for an emerging orthoflavivirus

Corresponding author: Anchun Cheng, chenganchun@vip.163.com

Corresponding author: Shun Chen, shunchen@sicau.edu.cn

a. Institute of Veterinary Medicine and Immunology, Sichuan Agricultural University, Chengdu, 611130, China;

b. Research Center of Avian Disease, College of Veterinary Medicine, Sichuan Agricultural University, Chengdu, 611130, China;

c. Key Laboratory of Animal Disease and Human Health of Sichuan Province, Sichuan Agricultural University, Chengdu, 611130, China;

d. Key Laboratory of Agricultural Bioinformatics, Ministry of Education, Chengdu, 611130, China;

e. Engineering Research Center of Southwest Animal Disease Prevention and Control Technology for Ministry of Education, Sichuan Agricultural University, Chengdu, 611130, China

Received Date: 04 January 2025

Accepted Date: 17 April 2025

Abstract: Highlights
1. A trans-complementation system for Usutu virus (USUV) recapitulates authentic viral replication.
2. High yield of TMN-ΔNS1 virus was obtained with no recombination into replication competent viruses.
3. TMN-ΔNS1 can be trans-complemented by heterologous orthoflavivirus NS1.

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一种新兴正黄病毒的非感染性假病毒系统构建

摘要

A noninfectious pseudovirus system for an emerging orthoflavivirus

Abstract

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Electronic Supplementary Material

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通讯作者: 陈斌, bchen63@163.com