rologie i - European Congress of Virology

5 th European Congress of Virologyin situ using Nile Red dye, which displayed a red emission for polar lipidsand a yellow emission for neutral lipids. The results showed that during thelytic phase (1 4 days after HHV8 infection) a depression of cholesterol andcholesterol ester synthesis (about 26%) was observed, while triglyceridesappeared to be enhanced up to 45% as compared to controls. After 14 24days, during the latent phase of infection, there was an increased synthesisof cholesterol (up to +44%) and cholesterol esters (up to +58%) in infectedcells, whereas triglycerides were progressively lowered. Imaging analysisshowed that the lipid droplets and total neutral lipid content were definitelyhigher (up to +85%) in HHV8 infected cells compared to the controls.Lipid synthesis inhibitors Sigma C75 and Sandoz 58035 were used toexamine the importance of lipid synthesis modification and neutral lipidaccumulation in the pathogenesis of KS and neo angiogenesis. This workwas financed by the Fondazione Banco di Sardegna 2012.REF 101Effect of Nitric Oxide on Mitochondrial Function in HepG2 cellsInfected by Dengue virusLeandro SILVA DA COSTA 1 , Holmes DE SOUZA CAMPOS 1 , ThaisMORAES DA CONCEIÇÃO 1 , Andrea THOMPSON DA POIAN 11 Universidade Federal do Rio de Janeiro Instituto de Bioquímica Médica,Rio de Janeiro, BRAZILDengue virus (DENV) infection causes dengue fever life threateningdengue hemorrhagic fever and dengue shock syndrome. Experimentalevidences suggest that liver is an important site of virus replication anddamage of this organ has been found in severe cases of DENV infection.It has been shown that human hepatocyte cell line, HepG2 producescytokines, and nitric oxide (NO) upon infection, which may be involvedin disease manifestations. In this work, we evaluated the effect of NOon DENV replication and mitochondria function in HepG2 cells. DENVreplication was evaluated by plaque assay and flow cytometry. NO productionwas investigated by intracellular NO quantification and inducibleNitric Oxide Synthase (iNOS) expression. Mitochondrial function wasmeasured in intact cells using high resolution respirometry. These resultsshowed an increase in NO production and in the expression iNOS and proapoptotic genes (BAX, Caspase 9) during the course of infection. Additionally,we demonstrated that infected cells present a significantly decreasein basal respiration and a 45% decrease in FCCP induced maximum electrontransport system capacity followed by apoptosis. When cells weretreated with NO inhibitor 1400w, these effects were abolished, suggestingNO involvement in mitochondrial alteration and apoptosis. We proposethat NO modulates mitochondria bioenergetics and dysfunction in DENVinfected cells. We believe that this study contribute to elucidate the molecularmechanisms underlying the role of NO in liver dysfunction causedby DENV. Financial Support: CNPq, FAPERJ.REF 103The Impact of Genetic Variations in IL28B Region on Response toTreatment of Chronic Hepatitis CIvana LAZAREVIC 1,2 , Jelena DJORDJEVIC 1,3 , Maja CUPIC 1,2 ,Danijela KARALIC 1,2 , Dragan DELIC 1,4 , Neda SVIRTLIH 1,4 , JasminaSIMONOVIC 1,4 , Petar SVORCAN 1,3 , Natasa MILIC 1,5 , TanjaJOVANOVIC 1,21 University of Belgrade, Faculty of Medicine, Belgrade, SERBIA;2 Institute of Microbiology and Immunology, Belgrade, SERBIA;3 Departement of Gastroenterology and Hepatology, Clinical Center“Zvezdara”, Belgrade, SERBIA; 4 Clinics of Infectious and TropicalDiseases, Clinical Center of Serbia, Belgrade, SERBIA; 5 Institute forMedical Statistics and Informatics, Belgrade, SERBIAThe three single nucleotide polymorphisms (SNPs) near IL28B gene wereshown to be highly predictive of sustained virological response (SVR)in patients with chronic hepatitis C virus (HCV) infection. The studyattempted to demonstrate the role of single and combined IL28B polymorphisms(rs8099917, rs12979860 and rs12980275) and other host andviral factors in predicting response to treatment, in Caucasian patientsinfected with HCV genotype 1. The study comprised 106 patients whounderwent standard 48 week therapy and out of which 55.7% achievedSVR. IL28B genotypes were determined by SSP PCR (Sequencespecific primer PCR). Patients carrying genotypes CCrs12979860 orAArs12980275 were 3.5 and 3 times more likely to achieve SVR, respectively.Genotypes GGrs8099917 and TTrs12979860 were identifiedas predictors of treatment failure. The presence of IL28B profile includingat least one of the favorable genotypes was identified as the mostimportant factor associated with SVR, followed by younger age andlower grade of histological activity. The strongest PPV of single SNPs forachieving SVR was observed for CCrs12979860 (76.9%). The presenceof GGrs8099917 showed the strongest NPV of 85.7%. The correlationof SNPs with other host and viral factors revealed association ofTTrs8099917 and lower AST levels. Results of this study confirm thatall investigated IL28B polymorphisms are associated with treatment responseand that presence of any of the favorable IL28B genotypes can beconsidered independent pretreatment determinant of the effectiveness oftherapy.REF 104Treatment outcome prediction model based on baseline viral geneticfactors and clinical variables in patients with chronic hepatitis CElisa MARTRÓ 1,2 , Verónica SALUDES 1,2 , Elisabet BASCUÑANA 1 ,Elena JORDANA LLUCH 1 , Sònia CASANOVAS 1 , Mercè ARDÈVOL 3 ,Esther SOLER 4,5 , Ramón PLANAS 4,5 , Vicente AUSINA 11 Microbiology Service, Hospital Germans Trias i Pujol, Badalona, SPAIN;2 CIBER Epidemiología y Salud Pública (CIBERESP), Barcelona, SPAIN.,Badalona, SPAIN; 3 Hospital Pharmacy, Hospital Universitari GermansTrias i Pujol, Badalona, SPAIN; 4 Liver Unit, Hospital UniversitariGermans Trias i Pujol; 5 CIBER Enfermedades Hepáticas y Digestivas(CIBEREHD), Barcelona, SPAINOnly about 50% of patients chronically infected with HCV genotype 1respond to treatment with pegylated interferon alfa (PegIFN a) and ribavirin(RBV), and protease inhibitors have to be administered together withthese drugs increasing costs and side effects. We aimed to develop a predictivemodel of treatment response based on a combination of baselineclinical and viral parameters. Seventy four patients chronically infectedwith HCV 1b and treated with PegIFN a and RBV were studied. Hostand viral related factors (viral load, and genetic variability in the E1 E2,core and ISDR regions) were assessed. Multivariate discriminant analysiswas used to develop a predictive model on the training group (N=53),which was then validated in the validation group (N=21). The predictivemodel generated included the following variables in decreasing order ofsignificance: number of viral variants in the E1 E2 region, an amino acidsubstitution pattern in the viral core region, the IL28B polymorphism,serum GGT and ALT levels, and viral load. Treatment outcome was accuratelypredicted in the training group (AUROC=0.9444; 96.3% specificity,94.7% PPV, 75% sensitivity, 81% NPV), and the accuracy remained highin the validation group (AUROC=0.8148, 88.9% specificity, 90.0% PPV,75.0% sensitivity, 72.7% NPV). The model obtained including host andviral factors had a high PPV in our population of Spanish HCV 1b treatmentnaïve patients. Accurately identifying those patients responding tothe dual therapy could help reducing implementation costs and side effectsof new treatment regimens.Virologie, Vol 17, supplément 2, septembre 2013S147