rologie i - European Congress of Virology

5 th European Congress of Virologyfever or dengue shock syndrome (DHF/DSS) due to endothelial cells andplatelets dysfunctions. Therapy is symptomatic and aims at controllingthe clinical manifestation of hemorrhages and shock. Prognosis of diseaseseverity is essential to adjust patient management. The pathogenesis of theDHF/DSS has not been fully elucidated but the nature of the virus hostinteractions has been identified as a potential risk factor. In order to analysevirion associated host protein composition, a method allowing the detailedanalyse of proteome of purified virions from HepG2 cells infected withdengue virus (DV) was developed. Five days after infection, virions werepurified from the culture supernatants by the combination of ultracentrifugationsand a water insoluble polyelectrolyte based technique. The purifiedparticles were controlled by immunoblots and electron microscopy. Afterin gel hydrolysis, peptides were analysed by nano liquid chromatographycoupled to ion trap mass spectrometer and identified on data libraries. Thelevel of viral NS1 protein excreted in the culture supernatants and thequantity of virus released were shown to be proportional to the viral input.Dengue virus structural proteins and more than 40 cellular proteins werespecifically and repeatedly identified on purified DV. The impact of theseproteins on disease outcome is discussed, as well as their potential use asfingerprint for dengue severity prognosis.Virologie, Vol 17, supplément 2, septembre 2013S169