模式识别受体ppt课件

.,1,InnateImmunerecognition,.,2,Threatstotheindividual,.,3,Protectionfrommicrobialinvasion,.,4,.,5,NobelPrize2011innateimmunity,.,6,NobelPrize2011,JulesA.Hoffmann:In1996,found“Toll”genehasaroleintheflysimmunitytofungalinfections.BruceA.Beutler:in1998,BruceA.BeutlerandcolleaguesdiscoveredthattheToll-likereceptors(TLRs)actastheprincipalsensorsofinfectioninmammals.RalphM.Steinman1973，RalphSteinmanisolateDentriticCell)。DCcanactivateTcell.DCisthebirdgebetweeninnateimmunityandadaptiveimmunity.,.,7,TollPathway,Drosophila,.,8,TheyalldeservetheNobelPrize,RuslanMedzhitov,CharlesJaneway19432003,TakashiFujita,ShizuoAkira,PRR,TLR,RLR,YaleUniv.,YaleUniv.,KyotoUniv.,OsakaUniv.,.,9,pathogenassociatedmolecularpatterns,PAMPsConservedpatternfoundonlyinpathogens,notinhostcells。(CharlesJenaway1989，atColdspringharbormeeting)TypicalPAMPs：1、Glycans：G-bacteria(lipopolysaccharide,LPS）;G+bacteria(peptidoglycan)；Bacterialflagellin2、Nucleotide:VirusDNA/RNA.,.,10,PAMPs,GlucanPeptidoglycanLPSFlagellin。,.,11,patternrecognitionreceptors,PRR：Receptors,onthesurfaceorinsidethecells,forthosepathogenassociatedmoleculepatterns.TypesofPRRs：Toll-likereceptors;TLRsRNAsensor（RIG-I,MDA5）DNAsensor（AIM2，DAI）NOD-likereceptors；NLRsC-LectinreceptorsOtherPRRs,.,12,PRR-PAMPsengagementactivatesphagocytes,.,13,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,.,14,1-1)CellulardistributionofTLRs,Cellmembrane,Endosomemembrane,.,15,PRRsinPMN,.,16,1-2)TLRsexpressionincells,ImmunecellsandsomeepitheliacellsexpressTLRs,.,17,TLRsexpressedinimmunecells,.,18,1-3)StructureofTLRs,*Homo-orheterodimers,ortogetherwithothermolecules,.,19,1-4)TLRssignalingpathway：,MyD88dependentpathway:AllTLRsexcept3MyD88independentpathway:TLR3;partofTLR4,effectorgenes,.,20,TLRssignalingpathway：,.,21,TLRssignalingpathway：,.,22,1.5)NegativeregulationofTLRssignaling,.,23,1-6)RoleofTLRsinimmuneresponse,Innateimmunity:Activatephagocytosisandkilling:regulatedexpressionofgenesofphagocytosisPromotingsecretionofcytokines/chemokines:IL1,IL6,TNFa,IFNs,CXCLs,CCLs(recruitotherimmunecells)Promotesecretionofanti-peptide:eg:Defensins,.,24,1-6)RoleofTLRsinimmuneresponse,Adaptiveimmunity:TLRsinduceDCmaturation:cytokinesecretion;costimulatorymolecules-DCactivation-presentantigentoactivateTcellsTLRssignalinginfluencesThcelldifferentiation:toinfluencethedifferentiationofTcells:Th1,Th2,Th17TLRsinduceTregactivation:regulatoryTcellsTregexpressTLR4/5/7/8,TLRsignaldirectlyinvolvedinthebiologicalfunctionofTregBcellactivation(eg:CpGDNATLR9ligandsmaybeusedasadjuvant),.,25,2-1）RNAsensors（TLRsandRLRs）,RIG-ILikeReceptors（RLR）：RIG-IMDA-5LGP2。,FujitaT,NatImmunol.2004JImmunol2005,.,26,2-2）structureofRIG-I,RIG-I结合5-pppdsRNA,.,27,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,.,28,2-3）RIG-1/MDA-5signalingpathway,.,29,2-4）regulatorofRIG-I/MDA-5,.,30,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,.,31,UbiquitinationforRIG-Isignaling,K63泛素化：激活K48泛素化：降解,Back,.,32,3-1）DNAsensor,DNAsensor：TLR9；PolIII；DAI；AIM2,.,33,4-1）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,.,34,4-2）NLRsactivtesinflammasome,PAMP通过NLRs激活炎症反应,.,35,NLRC4inflammasome,邵峰北京生命科学研究所ShaoF.(2011)Nature,477,596600.,.,36,5-1）C-Lectinreceptorfamily,Dectin-1，MannoseReceptors（MRs）,.,37,5-2）Thefungalpatternreceptor：Dectin-1,OnesuchsubfamilyistheDectin-1clusterwhichisprimarily,butnotexclusively,expressedbymyeloidcellsmacrophages,dendriticcells(DC)andneutrophils。,.,38,5-2）Thefungalpatternreceptor：Dectin-1,Back,.,39,DectinSignalingPathway,葡聚糖,甘露聚糖,甘露聚糖,.,40,6-2）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）：,.,41,CrosstalkofPRRsignaling,.,42,CrosstalkofPRRsignaling,.,43,PathogenevasionofPRRsignalingpathway,eg:HCVinteractwithPRRs,Fig1:structureofHCV,.,44,HCVinteractwithPRRs,.,45,PaperDiscussion（1）：RoleofTLR7inWNVinfection(TownTetal.,2009Immunity;IF20),ssRNAdsRNA(duringreplication),.,46,Fig1:IncreasedSusceptibilityofTlr7/andMyd88/Mice,butNotTlr9/Mice,afterWestNileVirusChallenge,WT:50%死亡TLR7-/-:90%死亡TLR7：抗病毒作用,.,47,Fig2.Tlr7-andMyd88-DependentViralLoadandInnateImmuneCytokineResponsesafterWestNileVirusChallenge：ViralLoad:Cytokine：IL23,IL12,.,48,Fig3：ImmuneCellHomingtoWNV-InfectedCellsInVivodependsonTLR7,Green：WNV（MoreinTLR7-/-）Red：ImmuneCells(LessinTLR6-/-),.,49,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)。,.,50,TLR7dependantmacrophagehoming,Lesscellinfiltration,.,51,Fig4.MacrophageHomingtoWestNileVirusIsIL-23SignalingDependent,Macrophage,Virus,HomingIssue,Brainsectionstaining,.,52,Summary:TLR7mediatedanti-WNVfunction,ImmuneCellHoming,VirusClearance,.,53,Paperdiscussion(2):Caspase-12controlsWestNilevirusinfectionviatheviralRNAreceptorRIG-I.(WangP.etal.,NatureImmunology2010),Caspase12-/-:Lesssurvival;Severeneurologicalsymptoms,.,54,1)ViralloadinCaspase12-/-mice,.,55,2)INFbproductioninCaspase12-/-mice,.,56,3)Caspase12interactswithRIG-I/TRIM25,.,57,4)LessUb-RIG-IinCaspase12-/-mice,Ub-RIG-I,TotalRIG-I,TotalTRIM25,.,58,StorySummary:Caspase12isrequiredforRIG-IUbIFNbproduction（Caspase12的抗病毒作用）,Casp12,eg:IFNproduction,.,59,PaperDiscussion（3）：Toll-likereceptor3mediatesWestNilevirusentryintothebraincausinglethalencephalitis（TLR3helpsthevirus?）WangT.etal.,2004NatureMedicine,WT:0%survivalTLR3-/-:40%survivalTLR3的存在增强了病毒感染,.,60,2）TLR3-/-micehaveviralloadinblood;butTNFa,IL6,.,61,3)TLR3-/-haveviralinbrain（TLR3-/-血液里virus多，但virus不能有效进入大脑。）,.,62,4)ReducedbraintissuedamageinTLR3-/-mice,炎性细胞渗入减少,.,63,5）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。,.,64,6）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,.,65,Summary：TLR3helpsWNVenterthebrainviaTNFa,*免疫分子往往是把双刃剑,血液-大脑,.,66,Paperdiscussion4:TLRandAging,JImmunol.2010Mar1;184(5):2518-27.PandaA,etal，YaleUniversitySchoolofMedicineAge-associateddecreaseinTLRfunctioninprimaryhumandendriticcellspredictsinfluenzavaccineresponse.,Fig1.TLRexpressioninDCs,.,67,Fig2.lesscytokineproductioninold,.,68,Fig3.Vaccineefficiencyinyoungandoldpopulation,VaccineefficiencyYoungold,Responsestoinuenzavaccinationinolderadultsandyoungadults.,.,69,Paper4Summary,1.LessTLRexpressioninold2.TLRsignalingarelessresponsiveinold3.DefectofTLRsignalingresultedinvaccinefailureinold,.,70,PaperDiscussion5:DDX24NegativelyRegulatesCytosolicRNA-MediatedInnateImmuneSignaling,.,71,.,72,DDX24interactwithFADD,.,73,IFNinduceDDX24expression,.,74,DDX24inhibitIFNproduction,.,75,SilenceDDX24，IFNproductionincreased,.,76,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,.,77,Figure4.SiRNA-mediatedknockdownofDDX24inhibitsVSVreplication.,.,78,Figure4.SiRNA-mediatedknockdownofDDX24inhibitsVSVreplication.,(F)LossofhDDX24affectsV