rologie i - European Congress of Virology

5 th European Congress of Virologythese enzyme activities and exhibited dual inhibition properties in the lowmicromolar range. Interestingly, amino acids mutations in the Non Nucleosideinhibitors binding pocket strongly affected the propenone derivativesRNase H inhibition, suggesting long range RT structural effects, withoutreducing their DNA polymerase inhibition ability. Thus we performedbiochemical and computational analyses in order to clarify their mode ofaction. Notably, these studies indicated that propenone derivatives are characterizedby a novel mechanism of action and bind to two different allostericinterdependent pockets. Hence, these results set the basis for the furtherdevelopment of RT inhibitors interacting with a novel RT binding site.REF 138Initial study on the mode of action of castalagin against herpes simplexvirusAngel S. GALABOV 1 , Neli VILHELMOVA 1 , Stephane QUIDEAU 21 The Stephan Angeloff Institute of Microbiology, Bulgarian Academyof Sciences, Sofia, BULGARIA; 2 Universite de Bordeaux, Institut desSciences Moleculaires, Pessak Cedex, FRANCEOur previous studies showed that castalagin, a nonahydroxyterphenoylbearing C glycosidic ellagitannin, manifests a significant antiviral activityagainst HSV 1 and HSV 2 strains both sensitive and resistant toACV. Moreover, the combination effect of ellagitannins with acyclovirwas shown as a markedly synergistic, especially the effect of this combinationtowards the replication of HSV 1 sensitive strain [Vilhelmova et al.,Antiviral Res. 89, 2011, 174 181; Antiviral Res. 90/2, 2011, N144, A64A65]. Here we present results of the initial step of a clearing up the modeof antiviral activity of castalagin on the replication of HSV 1 (Victoria,an ACV sensitive strain) in MDBK cells, including (i) timing of additionstudy in the one step growth cycle setup (MOI=1); (ii) the effect on theviral adsorption in MDBK cells, and (iii) the effect on the extracellularHSV virions. The susceptible to castalagin period of the virus groth cycleembraces markedly the initial first six hours post virus adsorption, as recordedat the 24th h: an inhibitory effect exceeding 3 logs when the compoundwas added immediately after or at the 1st and 2nd h post virus inoculation,and about 2 logs within the 3 h – 6 h time interval. The compound additionafter the 12th h manifested a negligible activity. Castalagin at the concentrationof 10 M (MTC) showed a marked effect on virus adsorption: log1.7 on the 30 min, 2.2 logs on the 45 min and 3.2 logs on the 60th min. Awell expressed inhibitory effect of 2 logs was registered by 1 M atthe60 min. At a concentration of 0.1 M effect the effect was clearly weaker(log 1) and at a concentration of 0.01 M a suppression of virus adsorptionwas not observed. The compound showed a marked direct inactivatingeffect on extracellular HSV 1, which is markedly temperature dependent.The effect was significantly higher at 37oC: castalagin 10 M decreasedthe infectious virus by log 1.6 at an exposure time of 30 min, 1.8 logs at60 min, 2 logs at 90 min, and 2.2 at the 120 min. This activity was stronglydose dependent and exposure time dependent: 30 min at the castalaginconcentrations of 10 M and 1 M, 60 min at 0.1 M and 0.01 M.REF 139Association study of IL28B: rs12979860 and rs8099917 polymorphismswith response/resistance to standard treatment of Moroccanspatients infected with hepatitis C virusNadia KANDOUSSI, Mohamed Rida TAGAJDID, BouchraBELFQUIH, Hicham ELANNAZ, Saâd MRANILaboratory of Virology, HMIM V, Faculty of Medicine and PharmacyMohamed V – Souissi University, Rabat, MOROCCOIntroduction: recently, four genome wide association studies (GWAS)have identified single nucleotide polymorphisms (SNPs) near the IL28Bgene (encoding IFN 3) to be strongly associated with spontaneous andtreatment induced clearance of HCV infection. The purpose of this studywas to investigate the association between two IL28B polymorphisms(rs8099917 and rs12979860) and response/resistance to standard treatmentof Moroccan patients infected with hepatitis C virus and allelic frequenciesof these polymorphisms answered most in a Moroccan population. Materialsand methods: this study was conducted on retrospective data of 265adult patients with chronic HCV infection who were diagnosed by antiHCV antibodies (ELISA). Data collection was performed with an Excelbased case report form laboratory, between January 2011 and December2012. All Patients had been treated with bitherapy IFN/RIBAVIRIN. 159patients had a sustained virologic response and 106 patients have persistentHCV viral load. Genotyping for the IL 28B rs12979860 C/T andrs8099917G/T polymorphisms was performed by High Resolution Meltingtechnical “HRM”. Early virological response (EVR) (12 weeks), endof treatment response (EOTR) (48 weeks), sustained virological response(SVR) (72 weeks) and relapse were evaluated using conventional andquantitative polymerase chain reaction (PCR) assays. Results and discussion:the frequency of the C allele in the population was 39%. Overall, 43%of patients experienced SVR. The IL28B CC genotype was significantlyassociated with higher treatment response rates and a lower relapse ratecompared to the other genotypes [84% vs. 58% EVR, 92% vs. 63% EOTR,76% vs. 38% SVR]. Thus, the IL28B genotype appears to be a strong predictorof SVR. This study, together with similar research examining otherSNPs, should help to define adequate protocols for the treatment of patientsinfected with HCV genotype 1, especially those with a poor prognosis.REF 140Prognostic role of Hepcidin level in Cchronic Hepatitis C virus patientson Interferon therapyNashwa KHADR, Hadier OKASHA, Hanan NOUH, Walaa SEMARYFaculty of Medicine University of Alexandria, Alexandria, EGYPTHCV is one of the most important health issues worldwide. Egypt has thehighest prevalence of hepatitis C in the world estimated about 15% 20%PEG–IFN plus ribavirin combination therapy became the gold standardin the treatment of HCV this treatment enhances sustained virologicalresponse (SVR)There are several host and viral factors aid in predictingresponse to treatment Hepcidine hormone is one of these host factors ithas a beneficial role in assessing treatment response during the courseof therapy Patients with chronic HCV often have increased liver iron, acondition associated with reduced response to antiviral therapy. The hepatichormone Hepcidine is the major negative regulator of iron release inthe circulation. The aim of the work is to assess the serum concentrationof hepcidin in chronic hepatitis C patients and evaluate any possibleassociation with the disease activity (viral load) as well as the efficacy ofpegylated IFN and Ribavirin therapy. This study was carried on 35 chronicHCV patients attending el kabary hospital to receive peg IFN/Ribavirintherapy and 15 chronic HCV patients not on IFN therapy as a control groupHepcidin hormone levels were measured in sera of 35 chronic hepatitis Cpatients before starting therapy (base line) at 12 and 24 weeks of therapyusing Hepcidine enzyme immuno assay (USCN life science Inc) Qualitativedetection of HCV RNA to asses sustained virologic response (SVR)using reverse transcriptase PCR at week 24 of antiviral therapy. Our casesshowed a male predominance (70%) over females (30%) and they showedthey showed a slight elevation of AST and Alkaline phosphatase while theother laboratory markers (ALT, Albumin, Bilirubin and Hb) were withinnormal values. The results of qualitative PCR for cases after 6 months oftherapy were in agreement with the RT-PCR results The level of hepcidinin all cases was very low before starting therapy and it showed a significantincrease during the course of therapy. This rise was detected earlyin responding cases. And it is significantly increased after 6 months oftherapy as compared to the control group. We found a negative correlationbetween baseline hepcidin level and baseline viral load of the cases thatshowed response after 6 months of therapy (undetectable viral RNA) wefound no significant relation between degree of fibrosis and viral load ofthe studied cases. Conclusion Chronic HCV infection is associated withVirologie, Vol 17, supplément 2, septembre 2013S157