Altered Protein Networks and Cellular Pathways in Severe West Nile Disease in Mice

Abstract

BackgroundThe 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.Methodology/Principal FindingsTo 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.Conclusion/SignificanceThis 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.

From <i>Culex</i> Exposure to West Nile Virus Infection: Screening of Specific Biomarkers

“…In order to better understand the pathophysiological processes in severe WNV infection and research early biomarkers of clinical course of WN disease, a recent proteomic study used animal model to this purpose. Potential protein biomarker candidates associated with the severity of WN disease clinical evolution are suggested such as High Mobility Group Box 1 (HMGB1) and Peroxiredoxin (PRDX6) [134]. This study has shown the kinetic modulation (the increased expression of HMGB1 and PRDX6 at the early and late time-points, respectively) of the inflammatory response leading to brain injuries following WNV infection [134].…”

“…At the protein level, proteomic profiling permited the identification of proteins whose expression levels were altered due to WNV infection in rat neurons [129] and Vero cells [132]. In addition, recently, proteomic study investigated the pathophysiological processes in severe WNV infection in mice, using kinetic analysis of protein expression profiles in the brain of WNV-infected mice prior and after the onset of clinical symptoms [134]. These proteomic studies in infected host cells revealed several host cellular functions that were altered following WNV infections: apoptotic pathway, cell cycle regulation or cytoskeleton maintenance associated with virus circulation, protein ubiquitination pathway, inflammatory response, and neurological development and neuronal cell death.…”

Possibilities of using proteomic analysis in infectiology

“…Чтобы получить представление о патофизиологических процессах, был изучен профиль белка в головном мозге мышей, инфицированных вирусом Çападного Нила. Исследования были проведены с использованием 2D-DIGE и iTRAQ методов с последующей идентификацией белков масс-спектрометрией [25]. С использованием протеомных методов было идентифицировано 148 ýкспрессированных белков в головном мозге мышей после инфицирования вирусом Çападного Нила.…”

“…С использованием протеомных методов было идентифицировано 148 ýкспрессированных белков в головном мозге мышей после инфицирования вирусом Çападного Нила. Эти исследования внесли огромный вклад в изучение патогенеза заболевания и разработки противовирусной терапии, что является основой для дальнейшего изучения биомаркеров заболевания, диагностики и профилактики заболеваний, вызванных вирусом Çападного Нила [25].…”

West Nile Virus Infections in (European) Birds

“…Understanding the correlates of virulence and pathogenesis of WNV neuroinvasive disease using stateof-the art technology could allow the identification of biomarkers and leads for novel treatment protocols. We have used the mouse model to study determinants of virulence [6,8] and to identify potential biomarkers [9]. Some of these markers have been validated using samples of human clinical cases [10].…”