Establishment of stable Vero cell lines expressing TMPRSS2 and MSPL: A useful tool for propagating porcine epidemic diarrhea virus in the absence of exogenous trypsin

Porcine epidemic diarrhea virus (PEDV) is the causative agent of porcine epidemic diarrhea, causing substantial economic losses to the swine industry worldwide. However, the development of PEDV vaccine is hampered by its low propagation titer in vitro, due to difficulty in adapting to the cells and...

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Autores principales:	Xiaona Wang, Xinyuan Qiao, Ling Sui, Haiyuan Zhao, Fengsai Li, Yan-Dong Tang, Wen Shi, Yuyao Guo, Yanping Jiang, Li Wang, Han Zhou, Lijie Tang, Yigang Xu, Yijing Li
Formato:	article
Lenguaje:	EN
Publicado:	Taylor & Francis Group 2020
Materias:	porcine epidemic diarrhea virus tmprss2 mspl vero cell lines isolation and propagation Infectious and parasitic diseases RC109-216
Acceso en línea:	https://doaj.org/article/101c1d273f4d4517a1e4c14338407f9b
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spelling	oai:doaj.org-article:101c1d273f4d4517a1e4c14338407f9b2021-11-17T14:21:58ZEstablishment of stable Vero cell lines expressing TMPRSS2 and MSPL: A useful tool for propagating porcine epidemic diarrhea virus in the absence of exogenous trypsin2150-55942150-560810.1080/21505594.2020.1770491https://doaj.org/article/101c1d273f4d4517a1e4c14338407f9b2020-12-01T00:00:00Zhttp://dx.doi.org/10.1080/21505594.2020.1770491https://doaj.org/toc/2150-5594https://doaj.org/toc/2150-5608Porcine epidemic diarrhea virus (PEDV) is the causative agent of porcine epidemic diarrhea, causing substantial economic losses to the swine industry worldwide. However, the development of PEDV vaccine is hampered by its low propagation titer in vitro, due to difficulty in adapting to the cells and complex culture conditions, even in the presence of trypsin. Furthermore, the frequent variation, recombination, and evolution of PEDV resulted in reemergence and vaccination failure. In this study, we established the Vero/TMPRSS2 and Vero/MSPL cell lines, constitutively expressing type II transmembrane serine protease TMPRSS2 and MSPL, in order to increase the stability and titer of PEDV culture and isolation in vitro. Our study revealed that the Vero/TMPRSS2, especially Vero/MSPL cell lines, can effectively facilitate the titer and multicycle replication of cell-adapted PEDV in the absence of exogenous trypsin, by cleaving and activating PEDV S protein. Furthermore, our results also highlighted that Vero/TMPRSS2 and Vero/MSPL cells can significantly enhance the isolation of PEDV from the clinical tissue samples as well as promote viral infection and replication by cell-cell fusion. The successful construction of the Vero/TMPRSS2 and Vero/MSPL cell lines provides a useful approach for the isolation and propagation of PEDV, simplification of virus culture, and large-scale production of industrial vaccine, and the cell lines are also an important system to research PEDV S protein cleaved by host protease.Xiaona WangXinyuan QiaoLing SuiHaiyuan ZhaoFengsai LiYan-Dong TangWen ShiYuyao GuoYanping JiangLi WangHan ZhouLijie TangYigang XuYijing LiTaylor & Francis Grouparticleporcine epidemic diarrhea virustmprss2msplvero cell linesisolation and propagationInfectious and parasitic diseasesRC109-216ENVirulence, Vol 11, Iss 1, Pp 669-685 (2020)
institution	DOAJ
collection	DOAJ
language	EN
topic	porcine epidemic diarrhea virus tmprss2 mspl vero cell lines isolation and propagation Infectious and parasitic diseases RC109-216
spellingShingle	porcine epidemic diarrhea virus tmprss2 mspl vero cell lines isolation and propagation Infectious and parasitic diseases RC109-216 Xiaona Wang Xinyuan Qiao Ling Sui Haiyuan Zhao Fengsai Li Yan-Dong Tang Wen Shi Yuyao Guo Yanping Jiang Li Wang Han Zhou Lijie Tang Yigang Xu Yijing Li Establishment of stable Vero cell lines expressing TMPRSS2 and MSPL: A useful tool for propagating porcine epidemic diarrhea virus in the absence of exogenous trypsin
description	Porcine epidemic diarrhea virus (PEDV) is the causative agent of porcine epidemic diarrhea, causing substantial economic losses to the swine industry worldwide. However, the development of PEDV vaccine is hampered by its low propagation titer in vitro, due to difficulty in adapting to the cells and complex culture conditions, even in the presence of trypsin. Furthermore, the frequent variation, recombination, and evolution of PEDV resulted in reemergence and vaccination failure. In this study, we established the Vero/TMPRSS2 and Vero/MSPL cell lines, constitutively expressing type II transmembrane serine protease TMPRSS2 and MSPL, in order to increase the stability and titer of PEDV culture and isolation in vitro. Our study revealed that the Vero/TMPRSS2, especially Vero/MSPL cell lines, can effectively facilitate the titer and multicycle replication of cell-adapted PEDV in the absence of exogenous trypsin, by cleaving and activating PEDV S protein. Furthermore, our results also highlighted that Vero/TMPRSS2 and Vero/MSPL cells can significantly enhance the isolation of PEDV from the clinical tissue samples as well as promote viral infection and replication by cell-cell fusion. The successful construction of the Vero/TMPRSS2 and Vero/MSPL cell lines provides a useful approach for the isolation and propagation of PEDV, simplification of virus culture, and large-scale production of industrial vaccine, and the cell lines are also an important system to research PEDV S protein cleaved by host protease.
format	article
author	Xiaona Wang Xinyuan Qiao Ling Sui Haiyuan Zhao Fengsai Li Yan-Dong Tang Wen Shi Yuyao Guo Yanping Jiang Li Wang Han Zhou Lijie Tang Yigang Xu Yijing Li
author_facet	Xiaona Wang Xinyuan Qiao Ling Sui Haiyuan Zhao Fengsai Li Yan-Dong Tang Wen Shi Yuyao Guo Yanping Jiang Li Wang Han Zhou Lijie Tang Yigang Xu Yijing Li
author_sort	Xiaona Wang
title	Establishment of stable Vero cell lines expressing TMPRSS2 and MSPL: A useful tool for propagating porcine epidemic diarrhea virus in the absence of exogenous trypsin
title_short	Establishment of stable Vero cell lines expressing TMPRSS2 and MSPL: A useful tool for propagating porcine epidemic diarrhea virus in the absence of exogenous trypsin
title_full	Establishment of stable Vero cell lines expressing TMPRSS2 and MSPL: A useful tool for propagating porcine epidemic diarrhea virus in the absence of exogenous trypsin
title_fullStr	Establishment of stable Vero cell lines expressing TMPRSS2 and MSPL: A useful tool for propagating porcine epidemic diarrhea virus in the absence of exogenous trypsin
title_full_unstemmed	Establishment of stable Vero cell lines expressing TMPRSS2 and MSPL: A useful tool for propagating porcine epidemic diarrhea virus in the absence of exogenous trypsin
title_sort	establishment of stable vero cell lines expressing tmprss2 and mspl: a useful tool for propagating porcine epidemic diarrhea virus in the absence of exogenous trypsin
publisher	Taylor & Francis Group
publishDate	2020
url	https://doaj.org/article/101c1d273f4d4517a1e4c14338407f9b
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Establishment of stable Vero cell lines expressing TMPRSS2 and MSPL: A useful tool for propagating porcine epidemic diarrhea virus in the absence of exogenous trypsin

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