5 th <strong>European</strong> <strong>Congress</strong> <strong>of</strong> <strong>Virology</strong>09. ANTIVIRAL THERAPY ANDRESISTANCE TO CHRONIC VIRALINFECTIONPosters: REF 130 to REF 146REF 130Humanized antibody targeting the envelope protein from HERV Wendogenous retrovirus in ongoing clinical trials for Multiple SclerosisHervé PERRON 1,2,3 , Jean Baptiste BERTRAND 2 , JaquesPORTOUKALIAN 3 , Reza FIROUZI 3 , Jack VANHORSSEN 4 , CorinneBERNARD 1 , Raphaëlle GERMI 5 , Patrice MORAND 5 , PatriceMARCHE 6 , Jean Louis TOURAINE 3 , Alois LANG 1 , FrançoisCURTIN 11 Geneuro, Geneva, SWITZERLAND; 2 Geneuro Innovation, Lyon,FRANCE; 3 Lyon1 Unviversity, Lyon, FRANCE; 4 VU University, Amsterdam,THE NETHERLANDS; 5 Joseph Fourier University & Hospital,Grenoble, FRANCE; 6 INSERM U823, Grenoble, FRANCEHERV W family retains elements expressing an envelope protein (Env),which activates inflammation through Toll Like receptor 4 (TLR4) onantigen presenting cells. HERV W/Env was evidenced by several independentRT PCR and immunohistological studies in MS brain lesions.About 75% <strong>of</strong> MS patients had Env antigenaemia in serum, as confirmedby quantitative RT PCR in blood lymphoid cells. Immunohistology<strong>of</strong> MS brain lesions showed expression <strong>of</strong> HERV W/Env in perivascularmacrophages and microglia. The animal model for MS, ExperimentalAllergic Encephalomyelitis, was induced by HERV W/Env in mice. Importantinflammatory demyelination was revealed by MRI and by histology.Anti myelin autoimmunity was also demonstrated. A Humanized antiHERV W/Env monoclonal antibody preventing activation <strong>of</strong> TLR4 byHERV W/Env displayed therapeutic and preventive effects in HERVW/Env induced mouse model. The Phase I and IIa clinical trials havenow been validated and the first assessment <strong>of</strong> a therapeutic agent targetinga human endogenous retroviral protein is ongoing in MS patients.This antibody <strong>of</strong>fers a strategic position in MS therapy, as (i) targetingan abnormally expressed immunopathogenic protein from an endogenousretrovirus, monitored by detection in patients, (ii) blocking the pathogeniceffect <strong>of</strong> this protein upstream the TLR 4 immune cascade activation, aswell as its direct effects on brain endothelial cells and on remyelinatingoligodendrocytes, and, (iii) without affecting the physiological functions<strong>of</strong> the human immune system as the majority <strong>of</strong> present treatments.REF 131The Efficacy <strong>of</strong> Colloidal Silver “ASAP” In Ameliorating Hepatitis BSurface Antigen Inffection Among Clinic Antendees in Abuja NigeriaOlu Joseph AJOBIEWE 1 , Semiyu OLAGOLDEN 2 , O. ODUNZE 21 NATIONAL HOSPITAL, ABUJA F.C.T, NIGERIA; 2 IMOSTATE UNI-VERSITY, OWERRI, NIGERIAIntroduction: several antiviral agents had failed to significantly clearhepatitis B Surface antigen in the systems <strong>of</strong> infected “ in” and “out”patients from our past clinical experience. This prompted us to test theeffect <strong>of</strong> Silver solution (ASAP) for this purpose. Study aim: to test theefficacy <strong>of</strong> colloidal Silver “ASAP” in ameliorating Hepatitis B Surfaceantigen infection among our clinic attendees. Study design/methods: thesetting <strong>of</strong> this prospective study was in Lugbe Federal Housing Estate inAbuja Municipal Area Council (AMAC) in Abuja, the federal capital city<strong>of</strong> Nigeria. The patients recruited for the study were mostly artisans andfew low cadre civil servants living in and around Lugbe and its environ.Sixty(60) people, making up <strong>of</strong> 40 men and 20 women were involved in thestudy who were highly reactive to the HbsAg serology test at the beginning<strong>of</strong> the study. They were also presenting with clinical signs and symptoms<strong>of</strong> hepatitis infection. Each <strong>of</strong> them was evaluated on various appointmentdays after silver solution (ASAP) 5mls per day was administered on them.Result: significant decline in reactivity to HbsAg serology test (p
5 th <strong>European</strong> <strong>Congress</strong> <strong>of</strong> <strong>Virology</strong><strong>of</strong> 17.3 per strain. At the protein level, 30 amino acid changes, correspondingto 22 different positions within pUS28, were identified, that is 6.2%<strong>of</strong> the total codons <strong>of</strong> the protein. Seven changes were located in putativefunctional motifs and 9 were unpreviously reported. All these polymorphismswere distributed evenly alongside pUS28. Overall, the frequency <strong>of</strong>these changes ranged from 2.4% to 56.1%. Among drug resistant viruses,22 out <strong>of</strong> the 30 polymorphisms found in drug sensitive viruses were evidencedtogether with 9 additional ones. Three novel polymorphisms werelocated in putative functional motifs. The distribution <strong>of</strong> US28 genotypeswas similar among drug sensitive and drug resistant viruses. In conclusion,the polymorphism <strong>of</strong> CMV US28 chemokine receptor is low and does notseem to be affected by CMV susceptibility to current antivirals.REF 134Genotypic characterization <strong>of</strong> human cytomegalovirus terminaseDavid BOUTOLLEAU 3 , Léa PILORGE 1 , Sonia BURREL 2,3 , ZaïnaAÏT ARKOUB 3 , Henri AGUT 2,31 Service de Vi<strong>rologie</strong>, Hôpital Pontchaillou, CHU de Rennes, Rennes,FRANCE; 2 UPMC Univ Paris 06, ER1 DETIV, Paris, FRANCE;3 Service de Vi<strong>rologie</strong>, Groupe Hospitalier Universitaire La Pitié Salpêtrière,CharlesFoix, Paris, FRANCEHuman cytomegalovirus (CMV) DNA packaging involves the CMV terminasecomposed <strong>of</strong> large UL56 and small UL89 subunits. Letermovir(AIC246), a novel anti CMV agent currently tested in phase II clinicaltrial, is able to interfere with CMV terminase. This work aimed to performthe genetic analysis <strong>of</strong> CMV terminase in genotypically characterized drugsensitive (n=33) and drug resistant (n=30) viruses. Full length CMV UL56and UL89 genes were amplified, sequenced, and compared to that <strong>of</strong> referencestrain AD169. Among drug sensitive viruses, interstrain identityvaried from 98 to 100% for both genes, with mean numbers <strong>of</strong> nucleotidemutations <strong>of</strong> 25.1 and 17.1 per strain for UL56 and UL89, respectively.At the amino acid level, 20 and 8 changes were identified in pUL56 andpUL89, respectively, that is 2.4% and 1.2% <strong>of</strong> the total codons <strong>of</strong> the proteins.Overall, the frequency <strong>of</strong> these changes ranged from 3% to 100%.Seven changes in pUL56, including 1 deletion, and 2 in pUL89 wereunpreviously reported. With regards to the putative functional domains<strong>of</strong> CMV terminase subunits, all amino acid changes were located outsideconserved regions, except S345A in pUL89. Conversely to pUL89,polymorphisms in pUL56 clustered mainly within two variable regions.Among drug resistant viruses, 19 out <strong>of</strong> the 28 polymorphisms foundin drug sensitive viruses were evidenced. Three changes were located inconserved domains (G649D and H698Q in pUL56, A659 V in pUL89).In conclusion, CMV terminase exhibits a low polymorphism and does notseem to be likely impacted by resistance to current antivirals.REF 135Genetic analysis <strong>of</strong> herpes simplex virus type 1 UL5/UL52 helicaseprimase complexSonia BURREL 2 , Marianne COLLOT 1,2 , Zaïna AIT ARKOUB 2 , HenriAGUT 1,2 , David BOUTOLLEAU 1,21 UPMC Univ Paris 06, ER1 DETIV, Paris, FRANCE; 2 Service de Vi<strong>rologie</strong>,Hôpitaux Universitaires La Pitié Salpêtrière Charles Foix, AP HP,Paris, FRANCEAmenamevir (ASP2151, Astellas Pharma) is a non nucleoside drug thatinhibits herpes simplex virus type 1 (HSV 1) UL5/UL56 helicase primase(HP) complex involved in viral DNA replication. Previous data evidencedsome amino acid changes associated with reduced susceptibilities <strong>of</strong> HSV1 to amenamevir in both subunits <strong>of</strong> HP complex that might alter viralreplication and pathogenicity. This work aimed to perform the genotypiccharacterization <strong>of</strong> HSV 1 HP complex among 26 drug sensitive isolates,6 acyclovir resistant isolates and KOS laboratory strain. Full length UL5and UL52 genes were amplified, sequenced, and compared to that <strong>of</strong> HSV1 reference strain 17. Among drug sensitive isolates, interstrain identityvaried from 99.4% to 99.9% for both genes, with mean numbers <strong>of</strong> nucleotidemutations <strong>of</strong> 8.9 and 12.4 per strain for UL5 and UL52, respectively.At the amino acid level, 21 and 26 changes were identified in pUL5 andpUL52, respectively, that is 2.4% and 2.5% <strong>of</strong> the total codons <strong>of</strong> theproteins. Of note, 1 isolate harbored a deletion at codons 695 696 betweenpUL52 conserved domains III and IV. For both proteins, all changesidentified lied within nonconserved regions, except V347I in pUL5. Theanalysis HP complex among 6 acyclovir resistant isolates showed polymorphismspreviously reported in drug sensitive isolates together with 6undescribed changes that were located outside functional and conservedregions, except M766 V in pUL52. This work confirms that UL5/UL52HP complex is highly conserved in HSV 1 clinical isolates and constitutesa legitimate antiviral target.REF 136Resistance development in HIV 1 infected individuals failing ARVtherapy: investigation <strong>of</strong> Gag and Pol polyproteins functional roleIlaria CARLI 1 , Claudia DEL VECCHIO 1 , Saverio Giuseppe PARISI 1 ,Federico DAL BELLO 1 , Arianna CALISTRI 1 , Giorgio PALÙ 1 , CristinaPAROLIN 21 Department <strong>of</strong> Molecular Medicine, Padua, ITALY; 2 Department <strong>of</strong> Biology,Padua, ITALYThe introduction <strong>of</strong> Antiretroviral Therapy (ARV) in the cure <strong>of</strong> AIDShas decreased the morbidity and mortality rate <strong>of</strong> HIV 1 positive patients.Nevertheless, since therapy is required lifelong, a significant minority <strong>of</strong>patients are likely to develop resistance to at least one class <strong>of</strong> drugs. Themechanisms <strong>of</strong> resistance mainly involve mutations altering the interaction<strong>of</strong> viral enzymes and inhibitors. Recent studies reveal that, besidesthe enzymes encoding ones, other regions might contribute to the development<strong>of</strong> resistance. In particular, some specific cleavage and non cleavagesite mutations in Gag increase polyprotein processing, thus compensatingfor the catalytic loss <strong>of</strong> the viral Protease (PR) functional activity inducedby primary resistance mutations. In this context, we are interested instudying the functional role <strong>of</strong> HIV 1 Gag in the viral life cycle <strong>of</strong> ARVselected viruses. We analyzed clinical samples <strong>of</strong> HIV 1 infected patientsfailing PR Inhibitors (PIs) and RT Inhibitors (RTIs) among a cohort <strong>of</strong>five infectious diseases units located in Veneto in northeastern Italy. Weoptimized PCR amplification and sequencing conditions for gag and polgenes. The relative contribution <strong>of</strong> Gag, PR and/or RT mutations on virusreplication ability and their interdependence are now examined using HIV1 molecular clones carrying patient derived sequences. Our results wouldcontribute to better characterize the role <strong>of</strong> Gag and the relations with PRand RT in resistance development, their relevance in viral replication andevolution in the presence or in the absence <strong>of</strong> drugs.REF 137Design, Synthesis and Biological Evaluation <strong>of</strong> New 1,3 Diarylpropenonesas Single Site Dual Inhibitors <strong>of</strong> HIV 1 RTFrancesca ESPOSITO 1 , Rita MELEDDU 1 , Valeria CANNAS 1 , SimonaDISTINTO 1 , Claudia DEL VECCHIO 2 , Giulia BIANCO 1 , AngelaCORONA 1 , Filippo COTTIGLIA 1 , Stefano ALCARO 3 , CristinaPAROLIN 4 , Elias MACCIONI 1 , Enzo TRAMONTANO 11 Department <strong>of</strong> Life and Environmental Sciences, University <strong>of</strong> Cagliari,Cagliari, ITALY; 2 Department <strong>of</strong> Molecular Medicine, University <strong>of</strong>Padova, Padova, ITALY; 3 Department “Salvatore Venuta”, University<strong>of</strong> Catanzaro, Catanzaro, ITALY; 4 Department <strong>of</strong> Biology, University <strong>of</strong>Padova, Padova, ITALYA small library <strong>of</strong> 1,3 diaryl propenones has been designed, and synthesizedas dual inhibitors <strong>of</strong> both HIV 1 reverse transcriptase DNA polymeraseand ribonuclease H associated functions. Compounds were assayed onS156 Vi<strong>rologie</strong>, Vol 17, supplément 2, septembre 2013
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