rologie i - European Congress of Virology

5 th European Congress of VirologyREF 125Resveratrol Inhibition of Human Rhinovirus ReplicationChiara NARDIS 1 , Chiara NARDIS 1 , Elena MATTIA 1 , Alessandra DELEO 1 , Antonio FRANCIOSO 2 , Luciana MOSCA 2 , PaolaMASTROMARINO 11 Department of Public Health and Infectious Diseases, Sapienza University,Rome, ITALY; 2 Department of Biochemical Sciences, SapienzaUniversity, Rome, ITALYRhinoviruses are the major etiological agents of common cold and asthmaexacerbations. However, no specific therapy is available. Resveratrol (RV)shows protective properties against several viral infections. We evaluatedthe antiviral effect of RV towards human rhinovirus 16 (HRV 16) in two differentcellular models: cell culture monolayer and human nasal epithelium,composed by basal, ciliated and mucus cells. Infection was established assingle or multiple cycles of virus replication. Viral yield in the presenceof different non cytopathic concentrations of RV was assessed by plaqueassay. In both cellular systems, RV showed a strong inhibitory effect onHRV 16 infective particles production, that appeared to be dose and timedependent. It has been reported that HRV 16 induces activation of NF kBwhich in turn promotes the transcription of ICAM 1, the major receptor forHRVs. We examined the effect of RV on the HRV 16 induced expressionof ICAM 1 and viral capsid proteins by immunofluorescence and westernblot assay. RV treatment inhibited viral protein synthesis and the HRV 16induced increase of ICAM 1 level. Since HRV 16 replication induces theproduction of pro inflammatory cytokines, we also evaluated the effects ofRV on IL 6, IL 8, and RANTES levels in human nasal epithelia by ELISAassay. HRV 16 induced increase of proinflammatory cytokines was reversedby RV treatment. These results suggest that RV may have a possibleclinical application in HRV infections, by reducing viral replication andthe related inflammatory symptoms.REF 126Antiviral Drug Design Platform (AD2P): screening of dengue virusinhibitorsJustine LETIENNE 1 , Cécilia EYDOUX 1 , Sabine MILHAS 1,3 , KarineALVAREZ 1 , Barbara SELISKO 1 , Xavier MORELLI 3 , GillesQUERAT 2 , Bruno CANARD 1 , Jean Claude GUILLEMOT 11 CNRS, AFMB, UMR 7257, ESIL, Marseille, FRANCE; 2 UMR 190,Unité des Virus Emergents, Faculté de Médecine de Marseille, Marseille,FRANCE; 3 Centre de Recherche en Cancérologie de Marseille (CRCM),CNRS UMR7258/INSERM U1068, Marseille, FRANCEWorldwide resurgence of dengue virus (DENV) infections is a majorconcern and few therapeutic treatments are available. The virus groupconsists of 4 serotypes and can induce similar symptoms ranging frommild febrile illness to a life threatening dengue hemorrhagic fever. Antiviralscreens have proved useful for the identification of novel humanemerging virus inhibitors. A screening workstation has been developed totest chemical compounds, based on the dynamic interface between threeplatforms: Interactions, Dynamics and Drug design platform (INT 3D);Screening platform Marseille Luminy (PCML) and Virology screeningplatform Marseille Timone (PCVMT). Our team (PCML) combines biochemicaland cell based screening assays to select efficient compounds(chemically synthesized or natural extracts). Our approach consists infocusing on non structural proteins of DENV that are essentials for viralreplication. A first strategy is to target non structural proteins NS5 and NS3to address the impact of drugs on enzymatic activities such as polymeraseor helicase activities. We are also interested in protein protein interactions(PPIs) virus network: viral (NS5/NS3) or cellular; in order to screenprotein protein interaction inhibitor (2P2I) library via HTRF or BRETassays. In addition, the data obtained in vitro are all validated in cellularcontext with a DENV replicon system and confirmed on cells infectedby dengue clinical isolates to assess inhibition effects and cytotoxicity ofcompounds. These strategies lead to the identification of promising DENVinhibitors.REF 127Polyclonal and monoclonal antibodies against human parechovirus 1and 3Brenda WESTERHUIS 1 , Ksm BENSCHOP 1 ,GKOEN 1 ,AqBAKKER 2 , Y CLAASSEN 2 , T BEAUMONT 2 , Kc WOLTHERS 11 Laboratory of Clinical Virology, Department of Medical Microbiology,Academic Medical Center, Amsterdam, THE NETHERLANDS; 2 AIMMTherapeutics, Academic Medical Center, Amsterdam, THE NETHER-LANDSThe immune response against picornaviruses is mainly humoral, andprotective neutralizing antibodies (Abs) are thought to be genotype specific.Human parechoviruses (HPeVs) are commonly detected picornavirusinfections in children. We observed that HPeV3 was difficult to neutralizein vitro, in disagreement with what is known for picornaviruses. The aimwas to study HPeV neutralization and develop neutralizing Abs againstHPeVs. Polyclonal Abs were obtained by rabbit immunization. Humanmonoclonal Abs were generated by generation of stable antibody producingB cells by genetic programming (AIMSelect). All Abs were testedfor neutralization in different cell lines. Polyclonal rabbit and monoclonalhuman antiHPeV1 Abs neutralized HPeV1 with high titers (>1:1024)and cross neutralization was found with HPeV2 (>512). In contrast, polyclonaland monoclonal aHPeV3 were not able to fully neutralize HPeV3infection in different cell lines. Although viral replication was inhibited tosome extent in the presence of Abs: the polyclonal aHPeV3 showed viralinhibition up to titer 1:256, and one monoclonal aHPeV3 inhibited replicationup to 1:32. In this study we generated highly neutralizing HPeV1Abs, unexpected both mAbs showed clear cross reactivity against HPeV2.Thus, cross neutralisation can occur in human picornaviruses, and therebypossibly cross protection in vivo as well. The generation of HPeV3 nAbswas difficult, and in line with our previous results. This finding needsfurther exploration.REF 128Mutation in the RNA-dependent RNA polymerase of Chikungunyavirus is responsible for resistance to T-705Nidya Segura GUERRERO 1,5 , Leen DEALNG 1 , Gilles QUERAT 2 ,Boris PASTORINO 2 , Mathy FROEYEN 6 , Kai DALLMEIER 1 , DirkJOCHMANS 1 , Felio BELLO 5 , Ali TAS 4 , Eric J. SNIJDER 4 , XavierDELAMBALLERIE 2 , Martina BRYON 3 , Johan NEYTS 1 , Martijn J.VAN HEMERT 4 , Pieter LEYSSEN 11 KU Leuven, Leuven, BELGIUM; 2 Université de la Méditerranée,Marseille, FRANCE; 3 Erasmus Medical Center, Rotterdam, THENETHERLANDS; 4 Leiden University Medical Center, Leiden, THENETHERLANDS; 5 Universidad del Rosario, Bogotá DC, COLOMBIAChikungunya virus (CHIKV) induced disease is an acute febrile illnesscharacterized by arthritis, arthralgia or severe joint pain that can persistfor several months up to years after the infection. There are no antiviraldrugs available for the treatment or prevention of CHIKV infections.Favipiravir, a nucleobase mimetic also known as T-705, is an effectiveand selective inhibitor of the replication of a broad spectrum of RNAviruses. We report here for the first time on the selective antiviral activityof T-705 on CHIKV replication and alphaviruses in general in tissueculture with median activities (EC50) in the range of 2-25 M observedfor lab as well as a panel of clinical strains of CHIKV. Additionally, firstproof-of-concept of the anti-CHIKV effect of T-705 is demonstrated invivo. In the interferon receptor-deficient (AG129) mice oral administrationof T-705 (300 mg/kg.day) reduced CHIKV induced mortality whenVirologie, Vol 17, supplément 2, septembre 2013S153