5 th <strong>European</strong> <strong>Congress</strong> <strong>of</strong> <strong>Virology</strong>Results: HPV was detectable from 258 cervix 39,9%, and from 92 analexamination 39,9%. 51% <strong>of</strong> cervix sample had only 1, genotype. Incontrast 15% <strong>of</strong> anal samples had only one 1, HPV genotype. 88,8% <strong>of</strong>anal samples HR, and 75,0% <strong>of</strong> anal samples had LR HPV genotype. FromHIV infected group 44% had other known sexually transmitted diseases(syphilis, gonorrhea, hepatitis B or C). Anal bleeding during sex (OR:1,66)fisting (OR1,18) passive role (OR:1,14) were independent risk factors forHR HPV after analysis. 57% <strong>of</strong> patients had more than 100 sexual partners.Conclusion: Hungarian MSM population is severely co infected with HPVand HR HPV. Smoking and high risk sexual behaviors are strong predictorfor acquiring HR HPV co infections. Early male vaccination programmight give the chance in preventing this potentially carcinogen infectionswab specimens. Epidemiologically, the typing results gave a comprehensivepicture <strong>of</strong> prevalent adenovirus genotypes during the four year periodin Finland.References1. Ylihärsilä M, Valta T, Hattara L, Karp M, Harju E, Hölsä J, Saviranta P,Waris M, Soukka T. Oligonucleotide array in well platform for detectionand genotyping human adenoviruses by utilizing upconverting phosphorlabel technology. Anal Chem 2011; 83: 1456-61.2. Ylihärsilä M, Harju E, Arppe R, Hattara L, Hölsä J, Saviranta P,Soukka T, Waris M. Genotyping <strong>of</strong> clinically relevant human adenovirusesby array in well hybridization assay. Clin Microbiol Infect 2012.doi: 10.1111/j.1469 0691.2012.03926.x.REF 506Epidemiological study <strong>of</strong> Human Adenovirus Genotypes in Finland,2009 2012Minna YLIHÄRSILÄ 1,2 , Tiina YLINEN 1 , Tero SOUKKA 2 , PetriSAVIRANTA 3 , Matti WARIS 11 Department <strong>of</strong> <strong>Virology</strong>, University <strong>of</strong> Turku, Turku, FINLAND;2 Department <strong>of</strong> Biotechnology, University <strong>of</strong> Turku, Turku, FINLAND;3 Medical Biotechnology Centre, VTT Technical Research Centre <strong>of</strong> Finland,Turku, FINLANDThere is scarce data on the epidemiology <strong>of</strong> human adenoviruses in Finland.In this study, an array in well hybridization assay [1,2] was used togenotype adenovirus types <strong>of</strong> cases detected in Finland between January2009 and December 2012. A total <strong>of</strong> 499 adenovirus positive respiratory,ocular swab, stool, and other types <strong>of</strong> specimens were included in thestudy. The specimens were first amplified with real time PCR based onthe adenovirus hexon gene, and the PCR products were typed by the arrayin well assay. The E04 (n=110, 22%), C02 (87, 17%) and C01 (46, 9%)genotypes were detected in every season. Overall, epidemic activity <strong>of</strong>type B03 (160, 32%) was observed in the autumn <strong>of</strong> 2010, after that onlysporadic cases were found. Genotypes D08, D19 and D37 (42, 8%), whichare known to cause epidemic keratoconjunctivitis, were detected in ocularREF 507ViralZone: recent updates to the virus knowledge resource.Patrick MASSON, Chantal HULO, Edouard DE CASTRO, LydieBOUGUELERET, Ioannis XENARIOS, Philippe LE MERCIERSuisse Institute <strong>of</strong> Bioinformatics, 1 rue Michel Servet, 1211 Genève 4,SuisseViralZone (viralzone.expasy.org) is a web resource that links sequencedata with virus biology knowledge, including virion structure, replicationcycle and host-virus interactions. The information is divided into viral factsheets for each viral genus, that describe virion shape, molecular biologyand epidemiology, with links to the corresponding sequences in GenBankand UniProt. Each viral genus page contains detailed illustrations, text,and PubMed references. This new update provides detailed view <strong>of</strong> viralmolecular biology through 133 viral ontology pages that describe commonsteps <strong>of</strong> viral replication cycles shared by several viral genera. Thisviral ontology is also represented in in the form <strong>of</strong> Gene ontology (GO)terms, and UniProtKB keywords. In this way, users can navigate from thedescription <strong>of</strong> a replication-cycle event to any viral genus concerned, andcan access to the sequences <strong>of</strong> genes related to this event.Vi<strong>rologie</strong>, Vol 17, supplément 2, septembre 2013S261
5 th <strong>European</strong> <strong>Congress</strong> <strong>of</strong> <strong>Virology</strong>25. RESPIRATORY VIRUS INFECTIONSPosters: REF 508 to REF 538REF 508Candidate plant derived universal Influenza A nanovaccines conferprotection against lethal homologous and heterologous virus challengePeter IVANOV, Tatiana GASANOVA, Natalia PETUKHOVALomonosov Moscow State University, Moscow, RUSSIAConservative Influenza M2e antigen (23 aa) has been overexpressed onthe surface <strong>of</strong> tobacco mosaic virus (TMV U1) particles. Three versions<strong>of</strong> M2e were inserted into the TMV coat protein (CP) gene. In additionto the consensus human M2e sequence, we introduced mutations by thesubstitution <strong>of</strong> cysteines with either serine or alanine residues. Agrobacteriummediated infections <strong>of</strong> Nicotiana benthamiana produced as much as4 g <strong>of</strong> TMV M2e ala per 1 kg <strong>of</strong> systemic leaves. Genetic stability <strong>of</strong> viralvectors was demonstrated. Chimeric virions consisted <strong>of</strong> two proteins andcontained up to 90% <strong>of</strong> CP M2e. Immunized mice did not lose weightcomparing with the control animals. The amount <strong>of</strong> antibodies specificto M2e exceeded those specific to TMV particle itself (maximum ratio5/1). Immunogold microscopy proved that the foreign antigens were uniformlydistributed and tightly packed on the surface <strong>of</strong> chimeric particles.The majority <strong>of</strong> TMV CP epitopes are hidden from the immune systemby the M2e antigens exposed on the particle surface. IgG1/IgG2a ratiovaries from 0.7 (Ser) to 3.2 (Ala). The anti M2e IgG response declinedinsignificantly during the 7 month period. Vaccinated mice were resistantto 5 LD50 <strong>of</strong> A/PR/8/34 (H1N1) and TMV M2e ala conferred partial protection(70% <strong>of</strong> survival rate) against heterologous strain (4 aa changes inM2e) influenza A/California/07/2009 (H1N1). Virus titer in lungs was twoorders lower than in the non immune animals. Therefore, a new generationcandidate universal Influenza A nanovaccines with specific rod shapedepitope geometry has been obtained.REF 509R.I.P. Rest in Peace: Human rhinoviruses target the death kinaseRIPK1 to interfere with caspase dependent cell deathMark LÖTZERICH, Urs F. GREBERInstitute <strong>of</strong> Molecular Life Sciences, University <strong>of</strong> Zurich, Zurich, SWIT-ZERLANDVirus infections can trigger apoptosis by activation <strong>of</strong> death receptorsor pattern recognition receptors, such as tumor necrosis factor receptor(TNFR), double strand RNA sensor toll like receptor 3 (TLR3) or RNAhelicase melanoma differentiation associated gene 5 (MDA5). Besides initiatorcaspase 8 and executor caspases this can involve regulatory kinases,such as receptor interacting protein kinase 1 (RIPK1). RIPK1 transmitsapoptotic signals via its death domain to caspase 8. We addressed howhuman rhinoviruses (HRV1a, 2, 14, 16, 37) interfere with cell death processesin Hela OHIO and primary human nasal epithelial cells. HRVinfection induced low levels <strong>of</strong> activation <strong>of</strong> caspases 8 and 9. Host chromatinrelocalized to the nuclear periphery and lamins were partially cleavedindependent <strong>of</strong> caspase activity. There was no sign <strong>of</strong> DNA fragmentation,poly ADP ribose polymerase cleavage, or activation cleavage <strong>of</strong> caspases3 and 7. Instead, the death domain <strong>of</strong> RIPK1 was cleaved to 60, 47 and37 kDa fragments. Ruptintrivir, an inhibitor <strong>of</strong> HRV 3C protease blockedthe formation <strong>of</strong> the 60 kDa RIPK1 fragment and abrogated cytopathiceffects. RIPK1 was found in a complex with 3C protease in infected cellsand recombinant 3C protease cleaved RIPK1. Collectively, the data suggestthat HRV induces upstream activation signals for apoptosis but blockexecution <strong>of</strong> apoptosis by 3C protease mediated cleavage <strong>of</strong> RIPK1. Thisleads to a non classical death pathway, which is controlled by viral genesrather than the host apoptosis machinery.REF 510Analyzing the receptor binding <strong>of</strong> influenza A viruses by using solublechimeric proteinsAnne Kathrin SAUER 1 , Chi Hui LIANG 2 , Maren BOHM 1 , ChristelSCHWEGMANN WESSELS 1 , Chung Yi WU 2 , Chi Huey WONG 2 ,Georg HERRLER 11 Institute <strong>of</strong> <strong>Virology</strong>, University <strong>of</strong> Veterinary Medicine, Hannover, GER-MANY; 2 Genomics Research Center, Academia Sinica, Taipei, TAIWANInfluenza A viruses can be found in many avian and mammalian speciesand are known for their ability to cross species barriers and adapt to newhosts. One major aspect for switching the host is the binding specificity<strong>of</strong> the surface glycoprotein hemagglutinin. It binds to cell surface sialicacids and viruses from different host species preferentially recognize sialicacids in certain linkage types. Avian influenza strains generally showa preference for 2,3 linked sialic acids whereas human strains show ahigher affinity to sialic acids linked in 2,6 conformation to galactose.Many studies use two plant lectins to stain for 2,3 and 2,6 linked sialicacids in cells and tissues: SNA (Sambucus nigra agglutinin) and MAA(Maackia amurensis agglutinin). But these two lectins have <strong>of</strong> course theirown individual binding properties. Using only these lectins is not sufficientto analyze the binding properties <strong>of</strong> HAs considering the high amount <strong>of</strong>different oligosaccharides even on a single cell and the variety <strong>of</strong> differentHA subtypes. To circumvent this problem we utilize soluble forms <strong>of</strong> severalHA subtypes to investigate the binding properties <strong>of</strong> the influenza HA,exploiting the receptor specificity <strong>of</strong> each subtype. Soluble hemagglutinins<strong>of</strong> the avian subtypes H5, H7 and H9 as well as human and porcineH1 subtypes have been tested on different cell lines and tissue sectionsfrom avian trachea and the porcine respiratory tract. Glycan array analysis<strong>of</strong> solHAs and lectins was performed to get a detailed insight into receptorbinding properties <strong>of</strong> influenza HAs.REF 511Efficient uncoating <strong>of</strong> influenza A virus requires exposure to mildlyacidic pH and a high concentration <strong>of</strong> potassium prior to fusion inlate endosomesSarah STAUFFER 1 , Yuehan FENG 1 , Firat NEBIOGLU 1 , LassiLILJEROOS 2 , Butcher SARAH J. 2 , Paola PICOTTI 1 , Ari HELENIUS 11 Institute <strong>of</strong> Biochemistry, ETH Zurich, Zurich, SWITZERLAND;2 Institute <strong>of</strong> Biotechnology, University <strong>of</strong> Helsinki, Helsinki, FINLANDFollowing endocytic uptake, Influenza A virus (IAV) is transported tolate endosomes where virus fusion is induced by the viral hemagglutinin,with an optimum <strong>of</strong> pH 5.0 for the strain X31 (A/Aichi/2/1968 (H3N2)).Acidification <strong>of</strong> the viral core in endosomes prior to fusion promotes efficientviral ribonucleoprotein (vRNP) uncoating. At mildly acidic pH theM2 channel mediates proton translocation into the viral core, resulting indisassembly <strong>of</strong> matrix protein 1 (M1) oligomers and release <strong>of</strong> vRNPsinto the cytoplasm following fusion. To date, core specific viral proteinprotein interactions and their dissociation upon acid exposure have mainlybeen analyzed in vitro. Here, we aimed to assess the pre fusion requirements<strong>of</strong> IAV uncoating in the context <strong>of</strong> infection, using pH 5.0 mediatedfusion at the plasma membrane. Mild acidification (pH 5.8) as well astreatment <strong>of</strong> IAV at high K+ concentrations mimicking the endosomalenvironment prior to fusion increased viral uncoating and infection significantly.In particular, core disassembly in vitro and in vivo, as monitoredby M1 dissociation and vRNP exposure, was facilitated when virions werepre acidified. We applied limited proteolysis combined with targeted massspectrometry in order to probe conformational changes in the virion. M1S262 Vi<strong>rologie</strong>, Vol 17, supplément 2, septembre 2013
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