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

Virulence

Wen Shi¹  Han Zhou²  Xiaona Wang²  Yuyao Guo²  Fengsai Li²  Yanping Jiang²  Li Wang²  Ling Sui²  Lijie Tang³  Xinyuan Qiao³  Yijing Li³  Yigang Xu³  Haiyuan Zhao⁴  Yan-Dong Tang⁵ 

[1] College of Animal Science and Technology, Northeast Agricultural University, Harbin, P.R. Chin;College of Veterinary Medicine, Northeast Agricultural University, Harbin, P.R. Chin;College of Veterinary Medicine, Northeast Agricultural University, Harbin, P.R. Chin;Heilongjiang Key Laboratory for Animal Disease Control and Pharmaceutical Development, Harbin, P.R. Chin;Department of Swine Breeding, Jiangsu Hanswine Food Co., Ltd, Ma’anshan, Anhui Province, Chin;State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin, Chin;

关键词: Porcine epidemic diarrhea virus;    TMPRSS2;    MSPL;    Vero cell lines;    isolation and propagation;   

DOI  :  10.1080/21505594.2020.1770491

来源: Taylor & Francis

【 摘 要 】

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.

【 授权许可】

CC BY