模式识别受体PPT参考幻灯片

InnateImmunerecognition,1,Threatstotheindividual,2,Protectionfrommicrobialinvasion,3,4,NobelPrize2011innateimmunity,5,NobelPrize2011,JulesA.Hoffmann:In1996,found“Toll”genehasaroleintheflysimmunitytofungalinfections.BruceA.Beutler:in1998,BruceA.BeutlerandcolleaguesdiscoveredthattheToll-likereceptors(TLRs)actastheprincipalsensorsofinfectioninmammals.RalphM.Steinman1973，RalphSteinmanisolateDentriticCell)。DCcanactivateTcell.DCisthebirdgebetweeninnateimmunityandadaptiveimmunity.,6,TollPathway,Drosophila,7,TheyalldeservetheNobelPrize,RuslanMedzhitov,CharlesJaneway19432003,TakashiFujita,ShizuoAkira,PRR,TLR,RLR,YaleUniv.,YaleUniv.,KyotoUniv.,OsakaUniv.,8,pathogenassociatedmolecularpatterns,PAMPsConservedpatternfoundonlyinpathogens,notinhostcells。(CharlesJenaway1989，atColdspringharbormeeting)TypicalPAMPs：1、Glycans：G-bacteria(lipopolysaccharide,LPS）;G+bacteria(peptidoglycan)；Bacterialflagellin2、Nucleotide:VirusDNA/RNA.,9,PAMPs,GlucanPeptidoglycanLPSFlagellin。,10,patternrecognitionreceptors,PRR：Receptors,onthesurfaceorinsidethecells,forthosepathogenassociatedmoleculepatterns.TypesofPRRs：Toll-likereceptors;TLRsRNAsensor（RIG-I,MDA5）DNAsensor（AIM2，DAI）NOD-likereceptors；NLRsC-LectinreceptorsOtherPRRs,11,PRR-PAMPsengagementactivatesphagocytes,12,TolllikeReceptor（TLR）,Nature.1997Jul24;388(6640):394-7.AhumanhomologueoftheDrosophilaTollproteinsignalsactivationofadaptiveimmunity.MedzhitovR,Preston-HurlburtP,JanewayCAJr.SourceSectionofImmunobiology,YaleUniversitySchoolofMedicine,NewHaven,Connecticut06520-8011,USA.AbstractWereportherethecloningandcharacterizationofahumanhomologueoftheDrosophilatollprotein(Toll)whichhasbeenshowntoinducetheinnateimmuneresponseinadultDrosophila.LikeDrosophilaToll,humanTollisatypeItransmembraneproteinwithanextracellulardomainconsistingofaleucine-richrepeat(LRR)domain,andacytoplasmicdomainhomologoustothecytoplasmicdomainofthehumaninterleukin(IL)-1receptor.BothDrosophilaTollandtheIL-1receptorareknowntosignalthroughtheNF-kappaBpathway.WeshowthataconstitutivelyactivemutantofhumanTolltransfectedintohumancelllinescaninducetheactivationofNF-kappaBandtheexpressionofNF-kappaB-controlledgenesfortheinflammatorycytokinesIL-1,IL-6andIL-8,aswellastheexpressionoftheco-stimulatorymoleculeB7.1,whichisrequiredfortheactivationofnaiveTcells.,PUBMEDsearch“Tolllikereceptor”24700papers,13,1-1)CellulardistributionofTLRs,Cellmembrane,Endosomemembrane,14,PRRsinPMN,15,1-2)TLRsexpressionincells,ImmunecellsandsomeepitheliacellsexpressTLRs,16,TLRsexpressedinimmunecells,17,1-3)StructureofTLRs,*Homo-orheterodimers,ortogetherwithothermolecules,18,1-4)TLRssignalingpathway：,MyD88dependentpathway:AllTLRsexcept3MyD88independentpathway:TLR3;partofTLR4,effectorgenes,19,TLRssignalingpathway：,20,TLRssignalingpathway：,21,1.5)NegativeregulationofTLRssignaling,22,1-6)RoleofTLRsinimmuneresponse,Innateimmunity:Activatephagocytosisandkilling:regulatedexpressionofgenesofphagocytosisPromotingsecretionofcytokines/chemokines:IL1,IL6,TNFa,IFNs,CXCLs,CCLs(recruitotherimmunecells)Promotesecretionofanti-peptide:eg:Defensins,23,1-6)RoleofTLRsinimmuneresponse,Adaptiveimmunity:TLRsinduceDCmaturation:cytokinesecretion;costimulatorymolecules-DCactivation-presentantigentoactivateTcellsTLRssignalinginfluencesThcelldifferentiation:toinfluencethedifferentiationofTcells:Th1,Th2,Th17TLRsinduceTregactivation:regulatoryTcellsTregexpressTLR4/5/7/8,TLRsignaldirectlyinvolvedinthebiologicalfunctionofTregBcellactivation(eg:CpGDNATLR9ligandsmaybeusedasadjuvant),24,2-1）RNAsensors（TLRsandRLRs）,RIG-ILikeReceptors（RLR）：RIG-IMDA-5LGP2。,FujitaT,NatImmunol.2004JImmunol2005,25,2-2）structureofRIG-I,RIG-I结合5-pppdsRNA,26,MAVS：adapterforRIG-I,Seth,R.B.,Sun,L.,Ea,C.,andChen,Z.J.,IdentificationandcharacterizationofMAVS:amitochondrialantiviralsignalingproteinthatactivatesNF-BandIRF3.Cell,122:669-682,2005,Prion-likeproteinfightoffviruses:MAVS,Cell2011,UniversityofTexasSouthwesternHHMI,27,2-3）RIG-1/MDA-5signalingpathway,28,2-4）regulatorofRIG-I/MDA-5,29,Ubiquitin(泛素）对PRR信号的调节,泛素由76个氨基酸残基组成，其中包括7个赖氨酸残基(K),其C末端可与底物的赖氨酸残基形成异肽键，从而引起底物泛素化。泛素的K11、K29、K48和K63均能参与形成泛素与泛素间的异肽键(Isopeptidebond)。,30,30,UbiquitinationforRIG-Isignaling,K63泛素化：激活K48泛素化：降解,Back,31,3-1）DNAsensor,DNAsensor：TLR9；PolIII；DAI；AIM2,32,4-1）NODlikereceptors（NLRs）,NLRs：C端：亮氨酸重复序列（LRR）中间：NACHT：寡聚体化，活化N端：CARD，PYD：相互作用,33,4-2）NLRsactivtesinflammasome,PAMP通过NLRs激活炎症反应,34,NLRC4inflammasome,邵峰北京生命科学研究所ShaoF.(2011)Nature,477,596600.,35,5-1）C-Lectinreceptorfamily,Dectin-1，MannoseReceptors（MRs）,36,5-2）Thefungalpatternreceptor：Dectin-1,OnesuchsubfamilyistheDectin-1clusterwhichisprimarily,butnotexclusively,expressedbymyeloidcellsmacrophages,dendriticcells(DC)andneutrophils。,37,5-2）Thefungalpatternreceptor：Dectin-1,Back,38,DectinSignalingPathway,葡聚糖,甘露聚糖,甘露聚糖,39,6-2）otherPRRs,清道夫受体（Scavengerreceptors）:结合脂类，LPS等甲酸基多肽受体（Formylpeptidereceptor)：结合含甲酸基的特殊氨基酸结构补体受体（Complementreceptor）：,40,CrosstalkofPRRsignaling,41,CrosstalkofPRRsignaling,42,PathogenevasionofPRRsignalingpathway,eg:HCVinteractwithPRRs,Fig1:structureofHCV,43,HCVinteractwithPRRs,44,PaperDiscussion（1）：RoleofTLR7inWNVinfection(TownTetal.,2009Immunity;IF20),ssRNAdsRNA(duringreplication),45,Fig1:IncreasedSusceptibilityofTlr7/andMyd88/Mice,butNotTlr9/Mice,afterWestNileVirusChallenge,WT:50%死亡TLR7-/-:90%死亡TLR7：抗病毒作用,46,Fig2.Tlr7-andMyd88-DependentViralLoadandInnateImmuneCytokineResponsesafterWestNileVirusChallenge：ViralLoad:Cytokine：IL23,IL12,47,Fig3：ImmuneCellHomingtoWNV-InfectedCellsInVivodependsonTLR7,Green：WNV（MoreinTLR7-/-）Red：ImmuneCells(LessinTLR6-/-),48,ImmuneCellHoming,Homing的意义：1、淋巴细胞得以合理的再分布及补充；2、增加抗原和淋巴细胞的接触机会，产生有效的免疫应答。(免疫细胞有效的迁移到受感染的部位)。,49,TLR7dependantmacrophagehoming,Lesscellinfiltration,50,Fig4.MacrophageHomingtoWestNileVirusIsIL-23SignalingDependent,Macrophage,Virus,HomingIssue,Brainsectionstaining,51,Summary:TLR7mediatedanti-WNVfunction,ImmuneCellHoming,VirusClearance,52,Paperdiscussion(2):Caspase-12controlsWestNilevirusinfectionviatheviralRNAreceptorRIG-I.(WangP.etal.,NatureImmunology2010),Caspase12-/-:Lesssurvival;Severeneurologicalsymptoms,53,1)ViralloadinCaspase12-/-mice,54,2)INFbproductioninCaspase12-/-mice,55,3)Caspase12interactswithRIG-I/TRIM25,56,4)LessUb-RIG-IinCaspase12-/-mice,Ub-RIG-I,TotalRIG-I,TotalTRIM25,57,StorySummary:Caspase12isrequiredforRIG-IUbIFNbproduction（Caspase12的抗病毒作用）,Casp12,eg:IFNproduction,58,PaperDiscussion（3）：Toll-likereceptor3mediatesWestNilevirusentryintothebraincausinglethalencephalitis（TLR3helpsthevirus?）WangT.etal.,2004NatureMedicine,WT:0%survivalTLR3-/-:40%survivalTLR3的存在增强了病毒感染,59,2）TLR3-/-micehaveviralloadinblood;butTNFa,IL6,60,3)TLR3-/-haveviralinbrain（TLR3-/-血液里virus多，但virus不能有效进入大脑。）,61,4)ReducedbraintissuedamageinTLR3-/-mice,炎性细胞渗入减少,62,5）BBBpermeabilityincreasedinWTbutinTLR3-/-miceafterWNVinfection,血脑屏障被打开,血脑屏障相对完好,Virus难以进入TLR3-/-小鼠脑部。,63,6）TNFaisimportantforBBBbreakdown,1）TNFa-/-鼠，WNV感染死亡率下降2）IL6-/-鼠，没差异3）TNFa-/-鼠，BBB保持相对完好TNFa是破坏BBB的细胞因子,64,Summary：TLR3helpsWNVenterthebrainviaTNFa,*免疫分子往往是把双刃剑,血液-大脑,65,Paperdiscussion4:TLRandAging,JImmunol.2010Mar1;184(5):2518-27.PandaA,etal，YaleUniversitySchoolofMedicineAge-associateddecreaseinTLRfunctioninprimaryhumandendriticcellspredictsinfluenzavaccineresponse.,Fig1.TLRexpressioninDCs,66,Fig2.lesscytokineproductioninold,67,Fig3.Vaccineefficiencyinyoungandoldpopulation,VaccineefficiencyYoungold,Responsestoinuenzavaccinationinolderadultsandyoungadults.,68,Paper4Summary,1.LessTLRexpressioninold2.TLRsignalingarelessresponsiveinold3.DefectofTLRsignalingresultedinvaccinefailureinold,69,PaperDiscussion5:DDX24NegativelyRegulatesCytosolicRNA-MediatedInnateImmuneSignaling,70,71,DDX24interactwithFADD,72,IFNinduceDDX24expression,73,DDX24inhibitIFNproduction,74,SilenceDDX24，IFNproductionincreased,75,Figure3SiRNA-mediatedknockdownofDDX24enhancesdsRNAinducedRLRsignaling.,(G)Genearrayindicatingmostup-regulatedgenesinMEFtreatedwithpolyI:Cat3hoursand9hoursVSnontreatment.Datafrom(A)(B)(C)(D)(E)(F)arepresentedasmeans6s.e.fromthreeindependentexperiments.*indicatesP,0.05.*indicatesP,0.01.doi:10.1371/journal.ppat.1003721.g003,76,Figure4.SiRNA-mediatedknockdownofDDX24inhibitsVSVreplication.,77,Figure4.SiRNA-mediatedknockdownofDDX24inhibitsVSVreplication.,(F)LossofhDDX24affectsVSVreplic