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Successive Passage In Vitro Led to Lower Virulence and Higher Titer of A Variant Porcine Epidemic Diarrhea Virus

文献类型： 外文期刊

作者： Zhao, Pengwei 1 ; Wang, Song 2 ; Chen, Zhi 3 ; Yu, Jiang 3 ; Tang, Rongzhi 3 ; Qiu, Wenbin 2 ; Zhao, Lu 4 ; Liu, Yueyue 4 ;

作者机构： 1.Zhejiang Univ, Sch Med, Dept Biochem & Mol Biol, Hangzhou 310058, Zhejiang, Peoples R China

2.Shandong Normal Univ, Ruminant Dis Res Ctr, Key Lab Anim Resistant Biol Shandong, Coll Life Sci, Jinan 250014, Shandong, Peoples R China

3.Shandong Acad Agr Sci, Shandong Prov Key Lab Anim Dis Control & Breeding, Jinan 250100, Shandong, Peoples R China

4.Shandong Acad Agr Sci, Shandong Key Lab Poultry Dis Diag & Immunol, Jinan 250023, Shandong, Peoples R China

5.China Inst Vet Drug Control, Beijing 100081, Peoples R China

6.Natl Agr Sci & Technol Ctr, Chengdu 610000, Sichuan, Peoples R China

关键词： porcine epidemic diarrhea virus; virus isolate; variant; pathogenicity; cytokines

期刊名称：VIRUSES-BASEL （ 影响因子：5.048； 五年影响因子：5.127 ）

ISSN：

年卷期： 2020 年 12 卷 4 期

页码：

收录情况： SCI

摘要： A highly virulent porcine epidemic diarrhea virus (PEDV) appeared in China and spread rapidly to neighbor countries, which have led to great economic losses to the pig industry. In the present study, we isolated a PEDV using Vero cells and serially propagated 100 passages. PEDV SDSX16 was characterized in vitro and in vivo. The viral titers increased to 10(7.6) TCID50/mL (100th) by serial passages. The spike (S) gene and the whole gene of the SDSX16 virus was fully sequenced to assess the genetic stability and relatedness to previously identified PEDV. Along with successive passage in vitro, there were 18 nucleotides (nt) deletion occurred in the spike (S) gene resulting in a deletion of six amino acids when the SDSX16 strain was passaged to the 64th generation, and this deletion was stable until the P100. However, the ORF1a/b, M, N, E, and ORF3 genes had only a few point mutations in amino acids and no deletions. According to growth kinetics experiments, the SDSX16 deletion strain significantly enhanced its replication in Vero cells since it was passaged to the 64th generation. The animal studies showed that PEDV SDSX16-P10 caused more severe diarrhea and vomiting, fecal shedding, and acute atrophic enteritis than SDSX16-P75, indicating that SDSX16-P10 is enteropathogenic in the natural host, and the pathogenicity of SDSX16 decreased with successive passage in vitro. However, SDSX16-P10 was found to cause lower levels of cytokine expression than SDSX16-P75 using real-time PCR and flow cytometry, such as IL1 beta, IL6, IFN-beta, TNF-alpha, indicating that SDSX16-P10 might inhibit the expression of cytokines. Our data indicated that successive passage in vitro resulted in virulent attenuation in vivo of the PEDV variant strain SDSX16.

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