丙型肝炎病毒ppt课件.ppt

HAV,HBV,HCV,HDV,HEV,黄热病病毒（YFV）,EB病毒（EBV）,巨细胞病毒（CMV）,单纯疱疹病毒（HSV）,风疹病毒（RV）,GBV-C（HGV）,TTV,病毒性肝炎viralhepatitis：由病毒感染引起的肝脏炎症病变,是一组主要感染肝细胞并在肝细胞中复制、增殖，并引起急、慢性肝脏炎症病变的病毒,HAV,HBV,HCV,HDV,HEV,肝炎病毒的定义：,0.05,34.1,抗-HBc,2011年度全国法定传染病报告发病情况(/10万),卫生部发布2011年度全国法定传染病疫情概况,病毒性肝炎发病情况,资料来源：卫生部法定传染病报告系统,急性丙肝,HCV感染的自然史,慢性丙肝（85%）,肝硬化（230%，20yrs）,失代偿（6%）,肝癌（4%）,死亡,死亡,36%,HepatitisCFromAtoZ,中国HCV感染的诊断和治疗现况,估计HCV感染者:500万1000万,2011年CDC报告发病数:173,872,仅占:1.6%3.2%,Vermehrenetal.DGVS2010;Gane.NewZealandDoctor.2007;/hep-c-facts-stats.html.,挑战：HCV感染绝大部分并未被诊断出来,1975,Descriptionofnon-A,non-Bhepatitis,1989,IdentificationofhepatitisCvirus,CourtesyofCharlieRice,HCV病毒学研究的重要时间节点,丙型肝炎病毒的发现,1970s:HepatitisAandBdiagnosticsMosttransfusionassociatedhepatitisnotcausedbyHAVandHBVorotherknownviralagents(NEJM292:767;Gastro.69:1278;JID139:511;VoxSang.44:231)non-A,non-Bhepatitis(NANBH)Mightbeasmall,envelopedvirustransmissibletochimpanzees(Gastro88:773;JID156:636)StandardimmunologicalmethodsfailedtoidentifyviralantibodiesorantigensChirongroupThoughtthattherewasnotenoughviralantigeninNANBHSotheydecidedtotryandconcentratethevirus,HightiterNANBHchimpanzeeplasma,Ultracentrifugationat104,000gfor5hours,Extractnucleiacid,randomprimedcDNAintogt11phage,ImmunoscreenedwithNANBHpatientserum,Clone5-1-1,LargeroverlappingClone81,丙型肝炎病毒的发现,PhagewithportionsofHCVgenome,InfectE.coli,PhagekillsE.coliandleavesplaques,ScreenwithHCVPatientserum,LibraryScreening,丙型肝炎病毒的发现,SouthernBlotswithClone81,Placenta,InfectedChimpLiver,AcuteChronic,Placenta,Clone81probe,IFN-probe,Clones5-1-1and81notfromhostgenomeandDNAreplicationintermediatesrelatedtotheseclonesnotdetected,NorthernBlotswithClone81,ClonesHybridizetoRNAfrominfectedanimals,RNAissinglestranded,sizeapprox5-10,000nucleotides,InfectedChimpLiver,UninfectedChimpLiver,UninfectedChimpLiver,2412g,DNaseI,RNaseA,Strand1Clone81probe,Strand2Clone81probe,ViralRNAds81,OligodT,unbound,Ctrl,Westernblot,Clone5-1-1expressedasSODfusionproteincalledPS5,PS5,SOD,NANBHpatientserum,Experimentallyinfectedchimpserum,Proteinfromclone5-1-1closelyassociatedwithNANBHinfections,HCVwasthefirstvirustobeisolatedandcharacterizedsolelybymolecularmethods,iewithoutculturemethods,黑猩猩易感,HCV病原学,HepatitisCGenomeRegionsCodingforStructural&Non-StructuralProteins,5Non-codingRegion,3Non-codingRegion,GenesCodingforStructuralProteins,GenesCodingforNon-Structural(NS)Proteins,HCVGenome,HepatitisCGenomeStructuralandNonstructuralGenes,C,A,NS2,NS3,E1,p7,E2,B,NS5,A,B,NS4,5Non-codingRegion,3Non-codingRegion,GenesCodingforStructuralProteins,GenesCodingforNon-Structural(NS)Proteins,HCVGenome,IRES,InternalRibosomalEntrySite(IRES)-ConsistsofDomainsII,III,IVofthe5NCRandfirst24-40nucleotidesofcoreregion-IRESinvolvedinprocessingofpolypeptideprecursor,HepatitisCVirusTranslation,HCVGenome,C,A,NS2,NS3,E1,p7,E2,B,5NCR,3NCR,NS5,A,B,NS4,PolyproteinPrecursor,Translation,HepatitisCVirusProteinProcessing,HCVGenome,C,A,NS2,NS3,E1,p7,E2,B,NS5,A,B,SignalPeptidase,NS2/3Protease,NS3/4AProtease,C,A,NS2,NS3,E1,p7,E2,B,NS5,A,B,NS4,PolyproteinPrecursor,ProteinProcessing,Translation,NS4,HepatitisCVirusProteinProcessing,HCVGenome,C,A,NS2,NS3,E1,p7,E2,B,NS5,A,B,NS4,PolyproteinPrecursor,ProteinProcessing,Translation,C,NS4B,NS5A,NS2,NS3,E1,NS4A,p7,E2,NS5B,Proteins,ProteinProcessing,HepatitisCGenomeTranslation,C,A,NS2,NS3,E1,p7,E2,B,NS5,A,B,NS4,PolyproteinPrecursor,SignalPeptidase,NS2/3Protease,NS3/4ASerineProtease,ProteinProcessingRoleofNS3/4ASerineProtease,C,NS4B,NS5A,NS2,NS3,E1,NS4A,p7,E2,NS5B,Proteins,HepatitisCVirusNS3/4ASerineProteaseInhibitors,C,A,NS2,NS3,E1,p7,E2,B,NS5,A,B,NS4,PolyproteinPrecursor,SignalPeptidase,NS2/3Protease,NS3/4ASerineProtease,NS3/4ASerineProteaseInhibitors,C,NS2,NS3,E1,p7,E2,Proteins,A,B,NS5,A,B,NS4,GenomicRNAisinfectious,HCVReplication,HCVLifeCycle,Typicalviruslifecycle:Binding,internalization,uncoating,proteinexpression/replication,assembly,release,HCVReplication-病毒的入侵需要细胞膜表面的受体,HCVreceptors/entryfactors,Intracellular,Linearpolysaccharidesonproteinsofallhumancellsurfaces,TetraspaninsuperfamilymemberExpressedinallnucleatedcellsPartofB-cellreceptorcomplex,ScavengerreceptorclassBmemberIHDLreceptor(multipleotherligands)Expressedinhepatocytes,adrenalcortex,gonads(lesselsewhere),FormthebackboneoftightjunctionstrandsinepithelialtissuesHighestexpressioninhepatocytes(lesselsewhere),Expressionconferssusceptibility,Silencingconfersresistance,Ligandsinfluenceinfection,Expressionconferssusceptibility,LDL-R,Lowdensitylipoproteinreceptor,(Agnelloetal.,1999),viralRNAaccumulationincreasedordecreasedinparallelwithLDLRmRNAexpressionandLDLentry,Providesawayforthevirustosticktocells,HCVReplication,AssemblyandReleaseCapsidproteininteractswith5NCRHomotypiccapsidproteininteractionE1andE2onERmembrane:NucleocapsidprecursorsbudintoERtoaquireenvelopePassthroughhostsecretorypathway,released,NatureReviewMicrobiology,-HCVcoreproteindrivesassemblyatthelipiddroplet（LD）-LDisboundbycore,thenNS5AandotherNSproteins,HCVReplication,Huh-7cellsinfectedbyhepatitisCvirus.Theexpressionofcoreproteinandactinfilaments,HCVleavesthecellapolipoproteinBsecretionpathway,DAA的临床研发,PawlotskyJM.H,HCVproteinsinteractwithmanyhost-cellfactorsandimpactonawiderangeofcellularactivities,HCVLifeCycle:KeyFeatures,MultipleproteinsmediateHCVentry:CD81,ScavengerReceptorB1,Claudin1,Occludin,VeryLowDensityLipoproteinReceptorInputHCVRNAistranslated,apolyproteinisformed,andindividualviralproteinsarereleasedfrompolyproteinbycellularandviralproteases,HCVLifeCycle:KeyFeatures,HCVproteinsassociatewithendoplasmicreticulummembranes,thesiteofHCVreplicationVirionassemblyoccursatlipiddropletsHCVleavesthecellbyhitchingarideontheapolipoproteinBsecretionpathwayHCVlifecycleisintimatelytiedwithlipidmetabolism,丙型肝炎流行病学WorldwidePrevalenceofHCVinfection,,全球性的健康问题(WHO)感染人数达2.27亿慢性感染者约1.7亿每年的新感染者达3万在北欧的发病率约为0.5%，而在埃及则高达20%估计仍有大量漏报病例（美国的“隐形杀手”）中国丙肝流行率?,HCVInfectionPrevalenceinChina,DaiZC,etal.TheviralhepatitisofChina:theepidemiologicalofserous,1997.60-71.,standardizedrate:Year1992-19953.2%38.4millionpatientsinChinaatleast,我国HCV感染流行率3.2%3千8百万感染者,HCVInfectionPrevalenceinChina,Recentdata:Anti-HCVingeneralpopulation:0.43%,我国HCV感染流行率最新数据0.43%500万感染者,PressRelease,MinistryofHealth,P.R.China.(2010).,Evolutionofanti-HCVantibodyassays,C,E1,E2,NS2,NS3,NS4B,p7,NS4A,NS5A,NS5B,structural,non-structural,c22,c33,c100,NS5,5-1-1,1stgenerationEIA,2ndgenerationEIA,3rdgenerationEIA/CIA,5-UTR,3-UTR,1990*,1992*,1996*,*YearsinwhichtheassayswereapprovedbyFDA,检测模式的改进,雅培Axysm罗氏Elecsys,雅培Architect,？,UnknownCauseHighprevalencePutian,Reuseofmedicalexperiments，56cases,Hailin,ContaminatedBloodtransfusion,64cases,Pingtang,ReportedEpidemicofHepatitisCinChina:2000-2010,HCVInfectionPrevalenceinChina,Recentdata:Anti-HCVingeneralpopulation:0.43%,我国HCV感染流行率最新数据1.2%5001000万感染者（校正后数据）,PressRelease,MinistryofHealth,P.R.China.(2010).,HCV变异,RNAvirus,RDRPlacksproof-readingfunctionMutationsariseduringreplicationarenotcorrectedRapidviralreplication:dailyreplicatedinTrillionGenotypesQuasispecies,HCV分为6个基因型中国以1b和2a基因型常见，1b型对IFN应答差基因6型主要见于香港和澳门,Quasispecies,HCV感染宿主后，经一定时期，在感染者体内形成以一个优势株为主的相关突变株病毒群，称为准种,HCVGenotypesandQuasispecies,Quasispeciesarekindoflikesiblings:highlyrelatedyetgeneticallydistinct,小结,HCVisamajorworldwidehealthconcernMuchremainsunknownaboutHCVCurrentdrugtherapyisinadequateandinsufficientNoveltherapiesofferIFN-resistantpatientsandthosewithserioussideeffectshopeofeliminationofhepatitisCinfection,核心抗原检测在HCV/HIV共感染人群的应用,HCViscommoninHIVpatients(approx25-40%inChinaandworldwide),HIV/HCV混合感染,PrevalenceofHCVinHIV+individuals:approx.90%inIVDU60-85%inhemophiliacs4-8%inHIV+MSM,HCVisamoreseriousdiseaseincoinfectedpatientsthaninmono-infected,HIV/HCV混合感染,HIVacceleratesHCVliverdisease:Within10-15yrsofinfection15-25%HIV/HCVcirrhosis2.6-6.5%ofthosewithoutHIVHCCappearstooccuratayoungerageAftershorterdurationofHCVinfection,HIVacceleratesHCVliverdiseaseespeciallywhenCD4declines,HIV/HCV混合感染,HIV感染慢性丙肝的筛查与诊断,HCV核心抗原定量检测在免疫缺陷人群的意义,实验设计,HCV核心抗原浓度HCVRNA水平抗HCV抗体浓度ALT、AST外周血CD4+T细胞计数,354份来自河南上蔡某自然村前有偿献血者，血清抗-HCV和/或抗-HIV抗体阳性。检测指标：,A：354份样本分为5组：HIV-CHC(129,36.44%)HIV+CHC(98,27.68%)HIV-SVC(65,18.36%)HIV+SVC(44,12.