rologie i - European Congress of Virology

5 th European Congress of Virologyresponse were overrepresented amongst the genes regulated in infectedcells. One of the highly upregulated genes in BAC16 RFPvIRF 3 infectedcells was TL1A. TL1A plays a role in costimulation of T cells and theTL1A/DR3 pathway is essential for the development of antiviral immunity,pointing to a novel mechanism of viral immune evasion by vIRF 3.Real time PCR targeting these miRNAs have been performed and confirmedthe differential expression pattern of these miRNAs at different timepoints following Coxsackie B4 virus infection. As a result, knowledge oftissue specific miRNAs expression during Coxsackie B virus replicationis essential to understand the aetiological role of virus induced T1D.REF 082Role of short telomeric repeat regions of Marek’s disease virus inreplication and lymphomagenesisAnnachiara GRECO, Nadine FESTER, Nikolaus OSTERRIEDER,Benedikt B. KAUFERInstitut für Virologie/Freie Universität Berlin, Berlin, GERMANYMarek’s disease virus (MDV) is a cell associated alphaherpesvirus thatcauses generalized polyneuritis and visceral lymphoma in chickens. MDVis able to integrate its genome into host telomeres. The MDV genome harborstwo telomeric regions at both termini: multiple telomeric repeats(mTMR), with a variable length, and a short telomeric repeat region(sTMR), with a fixed number of 6 repeats in all known MDV strains. Wepreviously demonstrated that mTMRs are dispensable for MDV replicationin vitro but play an important role in integration and tumor formationin vivo. To address the function of sTMRs in MDV replication and pathogenesis,we first deleted the entire sTMR sequence (vs TMR). vs TMRwas unable to replicate in vitro, indicating an essential role of the geneticelement. Furthermore, the minimal length of sTMR repeats necessary forMDV replication was determined using a set of sTMR truncation mutants.To address the role of the sTMR in MDV pathogenesis, we substituted theTMRs with a scrambled repeat sequence (vsTMRmut). vsTMRmut replicatedwith kinetics comparable to parental and revertant viruses in vitroand in vivo; however, disease and tumor incidence in vivo were severelyimpaired, confirming a function of sTMR in MDV pathogenesis. Takentogether our data suggest a central role of sTMR in MDV replication, likelyacting as spacer between the proximal packaging signal pac 1 and the DR 1cleavage site, as well as in MDV genome integration and tumor formation.REF 083Expression profile of microRNAs during Coxsackie B virus infectionin pancreatic cellsWai Yip LAM 1 , Apple Cm YEUNG 2 , Karry Lk NGAI 2 , Paul KsCHAN 2,3 , Stephen Kw TSUI 11 School of Biomedical Sceinces, Faculty of Medicine, The Chinese Universityof Hong Kong, Shatin, HONG KONG; 2 Department of Microbiology,Faculty of Medicine, The Chinese University of Hong Kong, Shatin, HONGKONG; 3 Stanley Ho Centre for Emerging Infectious Diseases, Faculty ofMedicine, The Chinese University of Hong Kong, Shatin, HONG KONGType 1 diabetes (T1D) is a disease characterized by the loss of insulinproducing beta cells in the pancreatic islets of Langerhans. It has beenreported that the incidence of T1D is increasing worldwide. MicroRNAs(miRNAs) are 21 to 23 nucleotides tissue specific non coding RNAs. MiR-NAs can bind to target sites in messenger RNAs with imperfect base pairingand, significantly reduce translational efficiency of the target proteins. Wehypothesize that Coxsackie B virus infection of beta cells may be inducingtowards the development of T1D. Furthermore, intact miRNA response inbeta cells are critical for the survival of beta cells and protection fromdiabetes subsequent to Coxsackie B infection. Pancreatic cells INS 1Eand RIN 5F cells were infected with Coxsackie B4 virus, respectively.MiRNAs were extracted and labeled using the Agilent miRNA labelingreagent. MiRNA microarray was used to delineate the expression profileof the miRNAs. Among the profiles of differentially regulated miRNA, itwas found that rno miR 375 and rno miR 382 were highly up regulatedor down regulated (> 3 fold) in both cell lines: RIN 5 and INS 1E, infectionwith Coxsackie B4 virus as compared with non infection controls.REF 084Cross talk between autophagic and apoptotic modulators in CCHFVinfected hepatocytesMarie MOROSO 1 , Olivier FERRARIS 2 , Raquel RODRIGUES 1 ,Christophe PEYREFITTE 2 , Glaucia PARANHOS BACCALA 11 Fondation Mérieux, Lyon, France; 2 Institut de Recherche Biomédicaledes Armées, Lyon, FRANCECrimean Congo hemorrhagic fever virus (CCHFV) is a tri segmentednegative stranded RNA virus belonging to the genus Nairovirus (Bunyaviridaefamily). It is transmitted to humans by ticks or direct contact withinfected blood and causes severe or even fatal hemorrhagic diseases inhumans. We demonstrated that hepatocytes are the main target cells ofCCHFV infection. To define the liver involvement in the pathogenesis,we analysed host responses induced by CCHFV in HuH7 and HepG2 celllines infected in vitro. HuH7 and HepG2 were shown to be permissiveto the virus and replicate it at a high viral load. In HuH7 infected cells,CCHFV elicited a cytopathogenic effect, but not in infected HepG2.Both CCHFV infected hepatocytes displayed common secreted IL 8. Notany other inflammatory cytokines such as TNF and IL 1 nor type I IFNwere detected. In addition, we observed a pro apoptotic CCHFV for bothhepatocytes but at different times.We found that CCHFV induced apoptosis in hepatocytes cells which wasmediated by the ER stress pathway, more characteristically the over expressionof CHOP and PERK mRNAs. Interestingly, CCHFV infected cellshad up regulated LC3B and p62 autophagic genes expression suggestingan activation of the autophagic machinery. We further decided to investigateat the protein level the autophagic markers. Detection of autophagicvesicles and LC3 protein conversion induced by CCHFV in the cross talkbetween apoptosis and autophagy will then be explained.REF 085Investigation of herpes simplex virus influence on male fertility: differentapproaches for different challengesVictor NAUMENKO 1 , Yurii TYULENEV 1 , Ekatherina MALOLINA 1 ,Andrey KULIBIN 2 , Elena GUSHCHINA 1 , Alla KUSHCH 11 Ivanovsky Institute of Virology, Moscow, RUSSIA; 2 Koltsov Institute ofDevelopmental Biology, Moscow, RUSSIAIntroduction: several studies have provided data for HSV impact in malesterility. At the same time it is not clear whether HSV is able to infectmature spermatozoa and what are pathogenic mechanisms of virus associatedsterility. The aim of this work was to develop several experimentalmodels for investigation of HSV influence on spermatogenesis and malefertility.Materials and methods: following objects were used for modeling ofHSV infection: human spermatozoa infected in vitro (model I), humantestis organotypic culture (model II), intraperitoneally infected prepuberalmice (model III), mature mice infected in rete testis (model IV). Rapidculture method, qPCR, electron and confocal microscopy were performedfor HSV detection and investigation of viral effects on spermatogenesis.Results: model I allowed to yield infected human spermatozoa in vitroand to reveal viral proteins in cytoplasm and acrosome. Using modelII it was shown that HSV was able to infect spermatogonia, spermatocytes,spermatides and it led to dramatic decrease of immature germ cellspopulation. Model III was useful to demonstrate that HSV could achievetestis before blood testis barrier formation. Mature mice that had beenS142 Virologie, Vol 17, supplément 2, septembre 2013