title: The effect of classical swine fever virus NS5A and NS5A mutants on oxidative stress and inflammatory response in swine testicular cells.
abstract: Infection with classical swine fever virus (CSFV) results in highly significant economic losses; this infection is characterized by being highly contagious and accompanied by hyperthermia and systemic bleeding. Oxidative stress (OS) plays a critical role in the pathological process of viral infection. The function of the nonstructural protein 5A (NS5A) in the pathogenesis of CSFV has not been completely understood. Here, OS and the inflammatory response were studied with NS5A and substitution mutants in swine testicular (ST) cells. ST cell lines stably expressing CSFV NS5A or substitution mutants were established. Reactive oxygen species (ROS) production, antioxidant protein expression and inflammatory response were analyzed by quantitative real-time PCR (qRT-PCR), ELISA and flow cytometry analysis. The results showed that CSFV NS5A did not increase ROS production or the antioxidant protein (Trx, HO-1 and PRDX-6) expression in ST cells. However, NS5A inhibited cyclooxygenase-2 (COX-2) expression, a pro-inflammatory protein related to OS. Further studies have shown that NS5A mutants S15A and S92A increased ROS production and inhibited antioxidant protein expression. S15A, S81A and T274A affected the inflammatory response. This study suggested that CSFV NS5A did not induce OS, and amino acids Ser15 and Ser92 of CSFV NS5A were essential for inhibiting OS. Additionally, Ser15, Ser81 and Thr274 played important roles in the inflammatory response in ST cells. These observations provided insight into the function of CSFV NS5A and the mechanism of CSFV persistent infection in ST cells.
authors.affiliation: College of Veterinary Medicine, Northwest A&F University, No. 22 Xinong Road, Yangling, 712100, Shaanxi, China.
authors.forename: Wang
authors.initials: W
authors.lastname: Dong
authors.affiliation: College of Veterinary Medicine, Northwest A&F University, No. 22 Xinong Road, Yangling, 712100, Shaanxi, China.
authors.forename: Huifang
authors.initials: H
authors.lastname: Lv
authors.affiliation: College of Veterinary Medicine, Northwest A&F University, No. 22 Xinong Road, Yangling, 712100, Shaanxi, China.
authors.forename: Yifan
authors.initials: Y
authors.lastname: Wang
authors.affiliation: College of Veterinary Medicine, Northwest A&F University, No. 22 Xinong Road, Yangling, 712100, Shaanxi, China.
authors.forename: Xiaomeng
authors.initials: X
authors.lastname: Li
authors.affiliation: College of Veterinary Medicine, Northwest A&F University, No. 22 Xinong Road, Yangling, 712100, Shaanxi, China.
authors.forename: Cheng
authors.initials: C
authors.lastname: Li
authors.affiliation: College of Veterinary Medicine, Northwest A&F University, No. 22 Xinong Road, Yangling, 712100, Shaanxi, China.
authors.forename: Lu
authors.initials: L
authors.lastname: Wang
authors.affiliation: College of Veterinary Medicine, Northwest A&F University, No. 22 Xinong Road, Yangling, 712100, Shaanxi, China.
authors.forename: Chengbao
authors.initials: C
authors.lastname: Wang
authors.affiliation: College of Veterinary Medicine, Northwest A&F University, No. 22 Xinong Road, Yangling, 712100, Shaanxi, China.
authors.forename: Kangkang
authors.initials: K
authors.lastname: Guo
authors.affiliation: College of Veterinary Medicine, Northwest A&F University, No. 22 Xinong Road, Yangling, 712100, Shaanxi, China. Electronic address: zhangym@nwsuaf.edu.cn.
authors.forename: Yanming
authors.initials: Y
authors.lastname: Zhang
mesh_terms: D000818:Animals; D002460:Cell Line; D006691:Classical Swine Fever / Q000473:pathology / Q000821:virology*; D006692:Classical Swine Fever Virus / Q000235:genetics*; D051546:Cyclooxygenase 2 / Q000378:metabolism; D007249:Inflammation / Q000378:metabolism / Q000662:veterinary*; D008297:Male; D009154:Mutation; D018384:Oxidative Stress; D017382:Reactive Oxygen Species / Q000378:metabolism; D060888:Real-Time Polymerase Chain Reaction; D013552:Swine; D013737:Testis / Q000166:cytology* / Q000821:virology; D017361:Viral Nonstructural Proteins / Q000235:genetics / Q000378:metabolism*
chemical_list: D017382:Reactive Oxygen Species; D017361:Viral Nonstructural Proteins; D051546:Cyclooxygenase 2
keywords: Classical swine fever virus; Inflammatory response; Mutants; NS5A; Oxidative stress