title: Cellular proteomic analysis of porcine circovirus type 2 and classical swine fever virus coinfection in porcine kidney-15 cells using isobaric tags for relative and absolute quantitation-coupled LC-MS/MS.
abstract: Viral coinfection or superinfection in host has caused public health concern and huge economic losses of farming industry. The influence of viral coinfection on cellular protein abundance is essential for viral pathogenesis. Based on a coinfection model for porcine circovirus type 2 (PCV2) and classical swine fever virus (CSFV) developed previously by our laboratory, isobaric tags for relative and absolute quantitation (iTRAQ)-coupled LC-MS/MS proteomic profiling was performed to explore the host cell responses to PCV2-CSFV coinfection. Totally, 3932 proteins were identified in three independent mass spectrometry analyses. Compared with uninfected cells, 304 proteins increased (fold change >1.2) and 198 decreased (fold change <0.833) their abundance in PCV2-infected cells (p < 0.05), 60 and 61 were more and less abundant in CSFV-infected cells, and 196 and 158 were more and less abundant, respectively in cells coinfected with PCV2 and CSFV. Representative differentially abundant proteins were validated by quantitative real-time PCR, Western blotting and confocal laser scanning microscopy. Bioinformatic analyses confirmed the dominant role of PCV2, and indicated that mitochondrial dysfunction, nuclear factor erythroid 2-related factor 2 (Nrf2)-mediated oxidative stress response and apoptosis signaling pathways might be the specifical targets during PCV2-CSFV coinfection.
authors.affiliation: Key Laboratory of Animal Virology of Ministry of Agriculture, College of Animal Sciences, Zhejiang University, Hangzhou, PR China.
authors.forename: Niu
authors.initials: N
authors.lastname: Zhou
authors.affiliation: Key Laboratory of Animal Virology of Ministry of Agriculture, College of Animal Sciences, Zhejiang University, Hangzhou, PR China.
authors.forename: Chunmei
authors.initials: C
authors.lastname: Fan
authors.affiliation: Key Laboratory of Animal Virology of Ministry of Agriculture, College of Animal Sciences, Zhejiang University, Hangzhou, PR China.
authors.forename: Song
authors.initials: S
authors.lastname: Liu
authors.affiliation: Key Laboratory of Animal Virology of Ministry of Agriculture, College of Animal Sciences, Zhejiang University, Hangzhou, PR China.
authors.forename: Jianwei
authors.initials: J
authors.lastname: Zhou
authors.affiliation: Key Laboratory of Animal Virology of Ministry of Agriculture, College of Animal Sciences, Zhejiang University, Hangzhou, PR China.
authors.forename: Yulan
authors.initials: Y
authors.lastname: Jin
authors.affiliation: Key Laboratory of Animal Virology of Ministry of Agriculture, College of Animal Sciences, Zhejiang University, Hangzhou, PR China.
authors.forename: Xiaojuan
authors.initials: X
authors.lastname: Zheng
authors.affiliation: China Institute of Veterinary Drug and Control, Beijing, PR China.
authors.forename: Qin
authors.initials: Q
authors.lastname: Wang
authors.affiliation: Institute of Animal Husbandry and Veterinary Medicine, Beijing Academy of Agriculture and Forestry Sciences, Beijing, PR China.
authors.forename: Jue
authors.initials: J
authors.lastname: Liu
authors.affiliation: College of Veterinary Medicine, China Agricultural University, Beijing, PR China.
authors.forename: Hanchun
authors.initials: H
authors.lastname: Yang
authors.affiliation: Key Laboratory of Animal Virology of Ministry of Agriculture, College of Animal Sciences, Zhejiang University, Hangzhou, PR China.|Institute of Immunology and College of Veterinary Medicine, Nanjing Agricultural University, Nanjing, PR China.
authors.forename: Jinyan
authors.initials: J
authors.lastname: Gu
authors.affiliation: Key Laboratory of Animal Virology of Ministry of Agriculture, College of Animal Sciences, Zhejiang University, Hangzhou, PR China.|State Key Laboratory and Collaborative Innovation Center for Diagnosis and Treatment of Infectious Diseases, First Affiliated Hospital, Zhejiang University, Hangzhou, PR China.
authors.forename: Jiyong
authors.initials: J
authors.lastname: Zhou
mesh_terms: D000818:Animals; D002460:Cell Line; D002853:Chromatography, Liquid / Q000379:methods*; D018173:Circoviridae Infections / Q000378:metabolism*; D017925:Circovirus / Q000737:chemistry* / Q000472:pathogenicity; D006691:Classical Swine Fever / Q000378:metabolism*; D006692:Classical Swine Fever Virus / Q000737:chemistry* / Q000472:pathogenicity; D016000:Cluster Analysis; D060085:Coinfection / Q000378:metabolism*; D007553:Isotope Labeling; D008954:Models, Biological; D013552:Swine; D053719:Tandem Mass Spectrometry / Q000379:methods; D014764:Viral Proteins / Q000032:analysis* / Q000737:chemistry / Q000378:metabolism
chemical_list: D014764:Viral Proteins
keywords: Classical swine fever virus; Coinfection; Mitochondrial dysfunction; Porcine circovirus type 2; iTRAQ