rologie i - European Congress of Virology

5 th European Congress of Virologyproteinases was registered in the isoform of the V th fraction. Conclusion:the V th isoform with high inhibitory activity was used for the study oftherapeutical properties and the experimental grippe A at white mice. Theisoform mentioned showed a promoted protective effect.REF 110A Rapid In Situ Enzyme Linked Immunosorbent Assay for DengueVirus Antiviral Marine Seaweed ScreeningClaudia DUARTE DOS SANTOS 1 , Andrea Cristine KOISHI 1 , PaulaRODRIGUES ZANELLO 1 , Éverson Miguel BIANCO 2 , JulianoBORDIGNON 11 Instituto Carlos Chagas/Fiocruz PR, Curitiba, BRASIL; 2 UniversidadeFederal de Santa Catarina, Florianópolis, BRASILDengue (DEN) is the most important mosquito borne viral disease in theworld. The total economic burden of dengue illness in the Americas wasestimated to cost US$ 2.1 billion per year on average. In Brazil it wasreported a total of 204,650 notifications of dengue and 324 severe caseswith 33 deaths in the first seven weeks of 2013. There is no vaccine orspecific antiviral therapy for prevention and treatment of DENV infection.Hence, drug discovery research for dengue is of utmost importance. Herewe propose a target free approach for dengue drug discovery based on anovel, rapid, and economic 96 well format in situ enzyme linked immunosorbentassay which can be adapted for high throughput screening. Thein situ ELISA was standardized for Huh 7.5 cell line and infection withall four serotypes of DENV, being DENV 1, 2 and 3 clinical isolates fromBrazil and DENV 4 a laboratory strain. The in situ ELISA was comparedto the foci forming assay and the results presented a high correlation (averager2 = 0.95), furthermore statistical analysis showed an average S/B of7.2 and Z factor of 0.62; demonstrating assay consistency and reliability.The assay was used to screen the antiviral activity of a panel of fifteen seaweedextracts at the maximum non toxic dose previously determined bythe MTT and neutral red cytotoxic assays. Eight seaweed extracts showeda dengue infection inhibition in the post infection treatment when comparedto the controls, with some variations depending on the virus serotype.Among these, four extracts were chosen for further evaluation. The preinfectiontreatment showed no significant inhibition of virus infection;on the other hand, treatment during the infection was highly effective,indicating that these extracts must interfere in early steps of the virusinfection cycle. Financial support: Fiocruz, CNPq, Fundação Araucaria,CAPES.REF 111Antiviral and cytoprotective activities of fullerenols with the differentcontent of Hydroxyl groupsMikhail EROPKIN 1 , Elena MELENEVSKAYA 2 , TatyanaBRYAZHIKOVA 1 , Elena EROPKINA 1 , Daria DANILENKO 11 Reserch Institute of Influenza, Saint Petersburg, RUSSIA; 2 Institute ofHigh Molecular Compounds, Saint Petersburg, RUSSIAWe have synthesized polyhydroxy fullerenes – fullerenols with thedifferent compound of hydroxyl groups – C60(OH)12 14, C60(OH)1824 and C60(OH)30 38. The first of them was non soluble in water andhad no biological activity when it was applied in cell cultures in formof a suspension. The two other variants of fullerenol possessed a broadspectrum of antiviral activity in vitro against the actual strains of humaninfluenza virus A(H1N1), A(H3N2) and avian influenza A(H5N1),human herpes simplex virus, adenovirus and respiratory syncitial virus.Water soluble variants of fullerenol were non toxic in vitro towardshuman and animal cells of the different tissue origin. Besides the antiviralactivity fullerenols demonstrated a protective effect against the UVAinduced phototoxicity. The maximum biological actitivy for all testedindices was shown in the fullerenol variant C60(OH)18 24. The watersoluble biologically active variants of fullerenol could be useful in thepharmacology since they could serve the basis for new effective and nontoxic antiviral and cytoprotective formulations.REF 112Consecutive Alternating Administration of Antiviral Combinationsas a Highly Effective Chemotherapy of Coxsackievirus B3 InfectionsAngel S. GALABOV, Ralitsa VASSILEVA PENCHEVAThe Stephan Angeloff Institute of Microbiology, Bulgarian Academy ofSciences, Sofia, BULGARIAEnteroviruses cause a great variety of diseases – from mild respiratoryand enteric infections to severe diseases of the CNS and the heart. Due toa rapid development of drug resistance of initially drug sensitive virusesanti enteroviral chemotherapeutics for clinical use are not registered sofar. One of the possible approaches to overcome this problem is by usingcombined chemotherapy. However, the application of a standard combinationtherapy, consisting of simultaneously given drugs could lead to thedevelopment of multiple resestance. The aim of the study, described in thecurrent paper is to apply a novel approach for combined administration ofanti enteroviral compounds, consisting of a consecutive application of thesubstances in experimental infection with Coxsackieviruses B3 in vivo.The used in vivo experimental models were neurotropic and cardiotropicCVB3 infection in newborn mice. Compounds partnering in combinationswere selected as specific inhibitors of enteroviral replication with differentmode of action disoxaril, guanidine HCl and oxoglaucine. The administrationof consecutively applied triple combination resulted in 40 60%survival rate of the treated with the combination mice, infected with lethalCVB3 infections. The effectiveness of the combination is also proven bysignificant lengthening of the mean survival time. Data presented in thecurrent study prove the effectiveness of previously chosen triple consecutivelyapplied combination of anti enteroviral inhibitors. By using suchcombination the occurence of drug resistance is totally avoided.REF 11310 23 DNAzyme targeted against the M2 gene transcript of influenzaA virus significantly inhibits viral RNA translation and replicationBinod KUMAR 1 , Prashant KUMAR 1 , Roopali RAJPUT 1 , LatikaSAXENA 1 , Dibyaranjan PATI 1 , Mradul DAGA 2 , Madhu KHANNA 11 Department of Respiratory Virology, Vallabhbhai Patel Chest Institute,University of Delhi, Delhi, INDIA; 2 Department of Medicine, MaulanaAzad Medical College, New Delhi, INDIAThe influenza A virus M2 ion channel protein is highly conserved amongdifferent viral strains and is essentially required during the trafficking,assembly and budding processes of virus, thus an attractive target fordesigning antiviral drugs. We designed several 10 23 DNAzymes (Dz)targeting different regions of the matrix gene (M2) of influenza A virusesand analyzed their ability to specifically cleave the target RNA in bothcell free systems as well as in cell culture using transient transfections.The Dz that worked best was further standardized with MgCl2 in a dosedependent manner. RT PCR and real time RT PCR assays showed significant75% inhibition of M2 gene of influenza A viruses upon specific Dztreatment. The transfection of MDCK cells with Dz considerably reducedthe cytopathic effect caused by influenza A virus (A/PR/8/34 H1N1) andconsiderably reduced the M2 protein expression. As expected, the mutantDz did not hinder in virus replication showing high level of specificity ofdesigned Dz towards the target RNA. Our results demonstrate substantialreduction in whole virus replication thereby paving new dimensionsin antiviral therapy. Our study, for the first time, has documented antiviralpotential of Dz against M2 transcript of influenza A virus. Thus, wepropose that the 10 23 DNAzymes may be used as selective and effectiveinhibitor of viral RNA replication, and can be explored further fordevelopment of a potent therapeutic agent against influenza infection.Virologie, Vol 17, supplément 2, septembre 2013S149