title: iTRAQ-based comparative proteomic analysis of Vero cells infected with virulent and CV777 vaccine strain-like strains of porcine epidemic diarrhea virus.
abstract: The re-emerging porcine epidemic diarrhea virus (PEDV) variant related diarrhea has been documented in China since late 2010 and now with global distribution. Currently, a virulent PEDV CH/YNKM-8/2013 and a CV777 vaccine strain-like AH-M have been successfully isolated from the clinical samples. To dissect out the underlying pathogenic mechanism of virulent PEDV and clarify the differences between virulent and CV777 vaccine strain-like PEDV infections, we performed an iTRAQ-based comparative quantitative proteomic study of Vero cells infected with both PEDV strains. A total of 661 and 474 differentially expressed proteins were identified upon virulent and CV777 vaccine strain-like isolates infection, respectively. Ingenuity Pathway Analysis was employed to investigate the canonical pathways and functional networks involved in both PEDV infections. Comprehensive studies have revealed that the PEDV virulent strain suppressed protein synthesis of Vero cells through down-regulating mTOR as well as its downstream targets 4EBP1 and p70S6K activities, which were validated by immunoblotting. In addition, the virulent strain could activate NF-κB pathway more intensively than the CV777 vaccine strain-like isolate, and elicit stronger inflammatory cascades as well. These data might provide new insights for elucidating the specific pathogenesis of PEDV infection, and pave the way for the development of effective therapeutic strategies.
  
  
  CONCLUSIONS
  Porcine epidemic diarrhea is now worldwide distributed and causing huge economic losses to swine industry. The immunomodulation and pathogenesis between PEDV and host, as well as the difference between virulent and attenuated strains of PEDV infections are still largely unknown. In this study, we presented for the first application of proteomic analysis to compare whole cellular protein alterations induced by virulent and CV777 vaccine strain-like PEDV infections, which might contribute to understand the pathogenesis of PEDV and anti-viral strategy development.
authors.affiliation: State Key Laboratory of Agricultural Microbiology, College of Veterinary Medicine, Huazhong Agricultural University, Wuhan 430070, China.
authors.forename: Xiaozhen
authors.initials: X
authors.lastname: Guo
authors.affiliation: State Key Laboratory of Agricultural Microbiology, College of Veterinary Medicine, Huazhong Agricultural University, Wuhan 430070, China.
authors.forename: Han
authors.initials: H
authors.lastname: Hu
authors.affiliation: State Key Laboratory of Agricultural Microbiology, College of Veterinary Medicine, Huazhong Agricultural University, Wuhan 430070, China.
authors.forename: Fangzhou
authors.initials: F
authors.lastname: Chen
authors.affiliation: State Key Laboratory of Agricultural Microbiology, College of Veterinary Medicine, Huazhong Agricultural University, Wuhan 430070, China.
authors.forename: Zhonghua
authors.initials: Z
authors.lastname: Li
authors.affiliation: State Key Laboratory of Agricultural Microbiology, College of Veterinary Medicine, Huazhong Agricultural University, Wuhan 430070, China.
authors.forename: Shiyi
authors.initials: S
authors.lastname: Ye
authors.affiliation: Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan 430070, China.
authors.forename: Shuang
authors.initials: S
authors.lastname: Cheng
authors.affiliation: State Key Laboratory of Agricultural Microbiology, College of Veterinary Medicine, Huazhong Agricultural University, Wuhan 430070, China.
authors.forename: Mengjia
authors.initials: M
authors.lastname: Zhang
authors.affiliation: State Key Laboratory of Agricultural Microbiology, College of Veterinary Medicine, Huazhong Agricultural University, Wuhan 430070, China. Electronic address: he628@mail.hzau.edu.cn.
authors.forename: Qigai
authors.initials: Q
authors.lastname: He
mesh_terms: D048868:Adaptor Proteins, Signal Transducing / Q000378:metabolism; D000818:Animals; D017209:Apoptosis; D002453:Cell Cycle; D002522:Chlorocebus aethiops; D002853:Chromatography, Liquid; D019295:Computational Biology; D020558:GTP Phosphohydrolases / Q000378:metabolism; D007249:Inflammation; D050356:Lipid Metabolism; D008856:Microscopy, Fluorescence; D016328:NF-kappa B / Q000378:metabolism; D010802:Phylogeny; D053485:Porcine epidemic diarrhea virus; D020543:Proteome / Q000378:metabolism*; D040901:Proteomics / Q000379:methods*; D012323:RNA Processing, Post-Transcriptional; D060888:Real-Time Polymerase Chain Reaction; D038762:Ribosomal Protein S6 Kinases, 70-kDa / Q000378:metabolism; D053719:Tandem Mass Spectrometry; D014709:Vero Cells / Q000378:metabolism / Q000821:virology*
chemical_list: D048868:Adaptor Proteins, Signal Transducing; D016328:NF-kappa B; D020543:Proteome; D038762:Ribosomal Protein S6 Kinases, 70-kDa; D020558:GTP Phosphohydrolases
keywords: Isobaric tags for relative and absolute quantitation (iTRAQ); Pathway analysis; Porcine epidemic diarrhea virus (PEDV); Proteomics