英文文献汇报_精品文档.ppt

英文文献汇报_精品文档.ppt

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IdentificationofalinearB-cellepitopeontheavianleukosisvirusP27proteinusingmonoclonalantibodies,XiaofeiLi1,2,LitingQin2,HaiboZhu3,YingjunSun2,XuezhiCui2,YadongGao2,XiaoleQi1,YongqiangWang1,HongleiGao1,YulongGao1,XiaomeiWang1ArchVirolIF=2.058,AbstractAvianleukosisvirus（ALV）isanavianoncogenicretrovirusthatcaninducevariousclinicaltumors.ThecapsidproteinP27isthegroup-specificantigenofALVandhasmanyviralantigensitesthatareeasytodetect.Inthisstudy,weproducedamonoclonalantibody（mAb）,3A9,thatisspecificfortheP27protein.Aseriesofpartiallyoverlappingpeptideswerescreenedtodefine181PPSAR185astheminimallinearepitoperecognizedbymAb3A9.Theidentifiedepitopecouldberecognizedbychickenanti-ALVandmouseanti-ALVP27sera.TheepitopewashighlyconservedamonganumberofALV-A,ALV-BandALV-Jstrains.MAb3A9mightbeavaluabletoolforthedevelopmentofnewimmunodiagnosticapproachesforALV,andthedefinedlinearepitopemighthelpfurtherourunderstandingoftheantigenicstructureoftheP27protein.,Avianleukosisviruses（ALVs）,whichbelongtothefamilyRetroviridae,areclassifiedintoA,B,C,D,E,andJsubgroups1,2.TheA,BandJsubgroupsofALVarethemostcommonALVsincommercialpoultry,whereassubgroupsCandDhaverarelybeenreported.SubgroupEisaubiquitousendogenousleukosisvirusoflowpathogenicity3.ALVpredominantlycauseslymphocyticleukemia,myeloidleukosisandothersarcomasandcanalsoleadtoimmunosuppressioneffects,suchastheabnormaldevelopmentofimmuneorgans,growthretardation,anddecreasedimmuneresponses.4,5.ALVsubgroupJ（ALV-J）wasfirstisolatedintheUKfromwhitemeat-typechickensin19882.InChina,ALV-Jwasfirstreportedin1999.Sincethen,ALV-Jisolateshavebeendetectedinlayerchickens,broilerbreedersandlocalchickensthroughoutmostregionsofChinaandhavecausedvarioustumorsinchickens68.Inrecentyears,ALV-Jhasbecomewidespreadandhasresultedinsevereeconomiclossestothepoultryindustry6,8.,1.Introduction,EffectivevaccinesagainstALVarenotavailable.ThecontrolofALVinfectionoccursprimarilybyestablishingexogenousALV-freepoultryflocksbyadoptingeradicationasthestrategyofchoice9.BecauseofsubstantialantigenicandgeneticvariationamongALVisolatesandhighlevelsofverticalandhorizontaltransmission,eradicationhasbeendifficult10,11.Thus,effectivemethodsfortheaccuratedetectionofALVantigensinchickensarecriticalforthecontrolofALVinfections.ThegenomeofALVconsistsof5-LTR-UTR-gag-polenv-UTR-LTR-3.Thegag,polandenvgenesaretheviralstructuralgenes11.TheP27protein,whichisencodedbythegaggene,isthecapsidproteinandactsasagroup-specificantigen.TheALVP27geneishighlyconservedandexhibits96%sequenceidentityamongtheexogenoussubgroups（A,B,C,DandJ）.Inaddition,theP27proteincontentaccountsformorethan30%oftheviralprotein,andP27hasmanyviralantigensitesthatareeasytodetect.Therefore,theP27proteinisthefirstchoiceforpreparingantibodiesfordetection.Epitopesofproteinsareclassifiedaseithercontinuousordiscontinuousdependingonwhethertheaminoacidsincludedintheepitopearecontinuousinthepeptidechainornot1214.Continuousepitopessometimesdonotrepresenttheentireantigenicepitopeintheviralproteinbutinsteadonlyashortcross-reactiveportionofalarger,discontinuousepitope12,13.,B-cellepitopesareregionsthatarerecognizedbythebindingsitesorparatopesofantibodymoleculeswhentheyarepresentintheirfreeforminserumorasmembrane-boundB-cellreceptors12.CorrectidentificationofB-cellepitopeswithinanantigenicproteinmayopenthedoorforthedesignofmoleculesthatmimicpotentiallyprotectiveepitopesandcouldbeusedtoraisespecificAbsorbeusedasprophylacticortherapeuticvaccines1517.IdentificationofB-cellepitopescouldpromoteprotectiveimmunityinthecontextofemergingandre-emerginginfectiousdiseasesandpotentialbioterroristthreats15.TheaimofthisstudywastogenerateaP27-specificantibodyagainstALVusingpurifiedrecombinantP27proteinastheimmunogenandsubsequentlytoidentifytheB-cellepitoperecognizedbytheantibody.TheinformationdescribedinthisstudywillfacilitatethedevelopmentofALVdiagnostictoolsandwillfurtherourunderstandingoftheantigenicstructureoftheP27protein.,2.Materialsandmethods,重组质粒的构建构建重叠片段短肽,免疫小鼠制备单抗,IFA&Westernblot检测单抗特异性,鉴定最小抗原表位基序,分析表位保守性,3.Results,Fig.1CharacterizationoftherecombinantP27proteinandmAb3A9.（A）ExpressionandpurificationoftherecombinantP27protein.M,molecularmarker（kDa）;1,recombinantP27proteinbeforeIPTGinduction;2,recombinantP27proteinafterIPTGinduction;3,purifiedrecombinantP27protein.（B）RecognitionofthepurifiedALV-JP27proteinbymAb3A9inwesternbl