title: Differential proteomic analysis of virus-enriched fractions obtained from plasma pools of patients with dengue fever or severe dengue.
abstract: BACKGROUND
  Dengue is the most widespread mosquito-borne viral disease of public health concern. In some patients, endothelial cell and platelet dysfunction lead to life-threatening hemorrhagic dengue fever or dengue shock syndrome. Prognostication of disease severity is urgently required to improve patient management. The pathogenesis of severe dengue has not been fully elucidated, and the role of host proteins associated with viral particles has received little exploration.
  
  
  METHODS
  The proteomes of virion-enriched fractions purified from plasma pools of patients with dengue fever or severe dengue were compared. Virions were purified by ultracentrifugation combined with a water-insoluble polyelectrolyte-based technique. Following in-gel hydrolysis, peptides were analyzed by nano-liquid chromatography coupled to ion trap mass spectrometry and identified using data libraries.
  
  
  RESULTS
  Both dengue fever and severe dengue viral-enriched fractions contained identifiable viral envelope proteins and host cellular proteins. Canonical pathway analysis revealed the identified host proteins are mainly involved in the coagulation cascade, complement pathway or acute phase response signaling pathway. Some host proteins were over- or under-represented in plasma from patients with severe dengue compared to patients with dengue fever. ELISAs were used to validate differential expression of a selection of identified host proteins in individual plasma samples of patients with dengue fever compared to patients with severe dengue. Among 22 host proteins tested, two could differentiate between dengue fever and severe dengue in two independent cohorts (olfactomedin-4: area under the curve (AUC), 0.958; and platelet factor-4: AUC, 0.836).
  
  
  CONCLUSIONS
  A novel technique of virion-enrichment from plasma has allowed to identify two host proteins that have prognostic value for classifying patients with acute dengue who are more likely to develop a severe dengue. The impact of these host proteins on pathogenicity and disease outcome are discussed.
authors.affiliation: MD3 Department, BioMerieux SA, 69280, Marcy l'Etoile, France. romain.fragnoud@hotmail.fr.
authors.forename: Romain
authors.initials: R
authors.lastname: Fragnoud
authors.affiliation: UMR 3569 de Virologie Structurale, Institut Pasteur, 75015, Paris, France. marie.flamand@pasteur.fr.
authors.forename: Marie
authors.initials: M
authors.lastname: Flamand
authors.affiliation: BioASTER, 60007, Lyon, France. frederic.reynier@bioaster.org.
authors.forename: Frederic
authors.initials: F
authors.lastname: Reynier
authors.affiliation: Unité de Virologie, Institut Pasteur Phnom-Penh, Phnom-Penh, Cambodia. pbuchy@pasteur-kh.org.
authors.forename: Philippe
authors.initials: P
authors.lastname: Buchy
authors.affiliation: Unité de Virologie, Institut Pasteur Phnom-Penh, Phnom-Penh, Cambodia. dvasna@pasteur-kh.org.
authors.forename: Vasna
authors.initials: V
authors.lastname: Duong
authors.affiliation: MD3 Department, BioMerieux SA, 69280, Marcy l'Etoile, France. alexandre.pachot@biomerieux.com.
authors.forename: Alexandre
authors.initials: A
authors.lastname: Pachot
authors.affiliation: Emerging Pathogens Laboratory Fondation Merieux, 69007, Lyon, France. glaucia.baccala@fondation-merieux.org.
authors.forename: Glaucia
authors.initials: G
authors.lastname: Paranhos-Baccala
authors.affiliation: MD3 & NAD Departments, BioMerieux SA, 69280, Marcy l'Etoile, France. frederic.bedin@biomerieux.com.
authors.forename: Frederic
authors.initials: F
authors.lastname: Bedin
mesh_terms: D000210:Acute-Phase Reaction; D000328:Adult; D003715:Dengue / Q000097:blood* / Q000821:virology; D003716:Dengue Virus / Q000235:genetics / Q000378:metabolism / Q000472:pathogenicity*; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D016179:Granulocyte Colony-Stimulating Factor / Q000097:blood; D054884:Host-Pathogen Interactions; D006801:Humans; D008297:Male; D010949:Plasma / Q000821:virology*; D010978:Platelet Factor 4 / Q000097:blood; D040901:Proteomics / Q000379:methods*; D019595:Severe Dengue / Q000097:blood / Q000821:virology; D053719:Tandem Mass Spectrometry / Q000379:methods; D014759:Viral Envelope Proteins / Q000032:analysis / Q000378:metabolism; D014771:Virion / Q000737:chemistry
chemical_list: C454127:OLFM4 protein, human; D014759:Viral Envelope Proteins; D016179:Granulocyte Colony-Stimulating Factor; D010978:Platelet Factor 4