title: Altered protein networks and cellular pathways in severe west nile disease in mice.
abstract: BACKGROUND
  The recent West Nile virus (WNV) outbreaks in developed countries, including Europe and the United States, have been associated with significantly higher neuropathology incidence and mortality rate than previously documented. The changing epidemiology, the constant risk of (re-)emergence of more virulent WNV strains, and the lack of effective human antiviral therapy or vaccines makes understanding the pathogenesis of severe disease a priority. Thus, to gain insight into the pathophysiological processes in severe WNV infection, a kinetic analysis of protein expression profiles in the brain of WNV-infected mice was conducted using samples prior to and after the onset of clinical symptoms.
  
  
  RESULTS
  To this end, 2D-DIGE and gel-free iTRAQ labeling approaches were combined, followed by protein identification by mass spectrometry. Using these quantitative proteomic approaches, a set of 148 proteins with modified abundance was identified. The bioinformatics analysis (Ingenuity Pathway Analysis) of each protein dataset originating from the different time-point comparisons revealed that four major functions were altered during the course of WNV-infection in mouse brain tissue: i) modification of cytoskeleton maintenance associated with virus circulation; ii) deregulation of the protein ubiquitination pathway; iii) modulation of the inflammatory response; and iv) alteration of neurological development and neuronal cell death. The differential regulation of selected host protein candidates as being representative of these biological processes were validated by western blotting using an original fluorescence-based method.
  
  
  CONCLUSIONS
  This study provides novel insights into the in vivo kinetic host reactions against WNV infection and the pathophysiologic processes involved, according to clinical symptoms. This work offers useful clues for anti-viral research and further evaluation of early biomarkers for the diagnosis and prevention of severe neurological disease caused by WNV.
authors.affiliation: Unité de Parasitologie, URMITE UM 63, Institut de Recherche Biomédicale des Armées (Armed Forces Biomedical Research Institute, IRBA), Marseille, France.
authors.forename: Christophe
authors.initials: C
authors.lastname: Fraisier
authors.forename: Luc
authors.initials: L
authors.lastname: Camoin
authors.forename: Stephanie M
authors.initials: SM
authors.lastname: Lim
authors.forename: Stéphanie
authors.initials: S
authors.lastname: Lim
authors.forename: Mahfoud
authors.initials: M
authors.lastname: Bakli
authors.forename: Maya
authors.initials: M
authors.lastname: Belghazi
authors.forename: Patrick
authors.initials: P
authors.lastname: Fourquet
authors.forename: Samuel
authors.initials: S
authors.lastname: Granjeaud
authors.forename: Ab D M E
authors.initials: AD
authors.lastname: Osterhaus
authors.forename: Penelope
authors.initials: P
authors.lastname: Koraka
authors.forename: Byron
authors.initials: B
authors.lastname: Martina
authors.forename: Lionel
authors.initials: L
authors.lastname: Almeras
mesh_terms: D000818:Animals; D001921:Brain / Q000821:virology; D002522:Chlorocebus aethiops; D005260:Female; D053858:Metabolic Networks and Pathways / Q000502:physiology*; D051379:Mice; D008810:Mice, Inbred C57BL; D012376:Rodent Diseases / Q000276:immunology / Q000378:metabolism* / Q000473:pathology; D012720:Severity of Illness Index; D057825:Two-Dimensional Difference Gel Electrophoresis; D014709:Vero Cells; D014901:West Nile Fever / Q000276:immunology / Q000378:metabolism* / Q000473:pathology / Q000662:veterinary; D014902:West Nile virus / Q000302:isolation & purification / Q000502:physiology