博士生课程2(2)-固有免疫模式识别课件

InnateImmunitymonocytemacrophagebacteriaThemostancientdefensePhysical&chemicalbarriersandcellularlineRecognitionbytheinnateimmunesystemsetsthestageforaneffectiveadaptiveimmuneresponse.机体在种系发生和进化过程中逐渐形成的一种天然免疫防御功能，构成机体抵御病原生物入侵的第一道防线.复习一、固有免疫系统的组成屏障细胞分子皮肤黏膜屏障：物理、化学、微生物血-脑屏障、血-胸腺屏障血-胎屏障、气-血屏障单核-巨噬细胞、中性粒细胞、树突状细胞、γδT细胞、NK细胞、NKT细胞、B1细胞、肥大细胞、嗜碱性粒细胞和嗜酸性粒细胞等。抗菌肽、溶菌酶、急性期蛋白、补体、细胞因子和黏附分子、Physical,chemicalandmicrobiologicalbarriersofourbody1、固有免疫屏障2、固有免疫细胞

PhagocyteNKILLs（固有样淋巴细胞）DCMCBasophil

Eosinophil

T细胞

NKT细胞

B1细胞Monocyte-macrophageNeutrophilRecognitionofaninfectiononceitgetspasttheepithelialbarrierPolarizationofTumor-associatedmacrophages(TAM)分泌

IL-1IL-6IL-12TNF-aIL-8GM-CSF细胞因子酶其它因子

杀伤肿瘤细胞

抗原呈递作用前列腺素白三烯补体成分纤维蛋白结合蛋白凝血因子溶菌酶酸性水解酶赖氨酸酶酯酶胶原蛋白酶弹性纤维蛋白酶免疫调节作用吞噬并杀伤病原微生物巨噬细胞的功能InteractionbetweenNeutrophilsandEndotheliumCellularAdhesionMolecules(CAMs):

Mucin-likeCAMsSelectinsIntegrinsIg-superfamily

CAMsNuocytesrepresentanewinnateeffectorleukocytethatmediatestype-2immunity.

Nature.2010Mar3.Type-2immunity:responsibleforprotectiveimmuneresponsestohelminthparasitesandtheunderlyingcauseofthepathogenesisofallergicasthma.Type-2cytokines:interleukinIL-4,IL-5andIL-13.Nuocytesexpandinvivoinresponsetothetype-2-inducingcytokinesIL-25andIL-33,andrepresentthepredominantearlysourceofIL-13duringhelminthinfection.InthecombinedabsenceofIL-25andIL-33signalling,nuocytesfailtoexpand,resultinginaseveredefectinwormexpulsionthatisrescuedbytheadoptivetransferofinvitroculturedwild-type,butnotIL-13-deficient,nuocytes.3、固有性免疫分子指体表分泌液以及血浆和其它体液中能够识别或攻击病原体的可溶性分子。抗菌肽antimicrobialpeptides溶菌酶lysozyme急性期蛋白(acutephaseproteins,APP)脂多糖结合蛋白（LBP）血清淀粉样蛋白（SAP）甘露糖结合蛋白（MBP）C反应蛋白等（CRP）补体细胞因子和黏附分子补体系统细胞因子和免疫相关细胞表面分子二、固有免疫识别病原相关分子模式(Pathogen-associatedmolecularpatterns,PAMPs)损伤相关分子模式（damage-associatedmolecularpatterns，DAMPs）模式识别受体（PatternRecognitionReceptors）病原相关分子模式(Pathogen-associatedmolecularpatterns,PAMP)：是病原微生物（尤其是原核生物）表面存在一些人体所没有的，但可为许多相关微生物所共享、结构恒定、进化保守的分子结构。PAMP的特征

1.通常为病原微生物所特有,乃天然免疫系统区分“自己”与“非己（微生物）”的分子基础。脂多糖：多数革兰阴性菌细胞壁成分；磷壁酸：多数革兰阳性菌胞壁成分；肽聚糖：革兰阳性/阴性菌、真菌胞壁成分；甘露糖：微生物细胞壁上糖蛋白和糖脂成分2.为微生物生存和致病性所必需

PAMP突变或缺失微生物死亡或微生物对外界环境适应性3.宿主泛特异性识别的分子基础

PAMP是由一群或一类特定的微生物所共有的恒定结构（如LPS）。宿主由种系编码的有限数量PRR可察觉任何微生物感染的存在Pattern

Lipopolysaccharide(LPS)

Lipoteichoicacid

Bacteriallipopeptides

Peptidoglycan

Yeastandgram+bacteria

BacterialDNA(CpG)

Flagellin

Terminalmannose/fucose

ViralDNA(CpG)

ssRNA

dsRNAPathogen-

Associated

Molecular

Patterns

(PAMP)损伤相关分子模式（damage-associatedmolecularpatterns，DAMPs）机体自身细胞所释放的内源性分子，即内源性危险信号，来源于受损或坏死组织和某些激活的免疫细胞。主要有HMGB1、热体克蛋白等。PAMPvsDAMPSterileinflammationconservedmicrobialmotifsVSnon-microbialsignalsEXTRACELLULAR/SECRETEDPRRsMannosebindinglectin/protein(MBP)C-reactiveprotein(CRP)Serumamyloidprotein(SAP)LPS-bindingprotein(LBP)AcutephaseproteinsAcutephaseresponse(APR):theserumchangesAPRproteins:theirconcentrationsroseorfell(duringtheacutephase)Sitesofinjuryorinfectionsignals(proinflammatorycytokines:TNF-,IL-1,&IL-6producedbyphagocytes)stimulatingLiver:synthesisofAPRproteinsIncreaseinthelevelofC-reactiveprotein&Mannose-bindinglectin/MBL&

Serumamyloidprotein/SAP&

fibrinogen)MBLMannosebindinglectinLungsurfactantsA,DFicolins“patternrecognitionreceptors”;inthiscasepatternofterminalsugarsoncellsurfacesRecognizingmannose-containingmolecularpatternsfoundonmicrobesbutnotonvertebratecells

directing

complementattack

Mannose-bindinglectinMannosebindingprotein(MBP)PartofC-typelectinsuperfamilyAssociateswithandactivatesserineproteases:MASP-1andMASP-2Afterbindingtopathogensurfacethiscomplexactivateslectinpathwayofcomplementsystem,C2andC4MB-LECTINMASP=MBL-associatedserineproteaseMBLmannoseANOTHERVERSIONPOINTSOUTTOTALITYOFCLEAVEDC3FUNCTIONSBindtophorphorylcholine(PC)onbacteria,othermicroorganisms,damagedhostcellmembranesPCfoundinteichoicacids,capsularcarbohydrates,andlipopolysaccharidesRequiresCa++Functiondirectlyasopsonins(enhancerofphagocytosis)FunctionindirectlybybindingtoC1qofclassicalcomplementpathwayandactivatecomplementcascadeC-reactiveprotein(CRP)andserumamyloidprotein(SAP)(belongstoafamilyofpentamericproteincalledpentraxins)bindingto

polysaccharide&phophorylcholine(=ligands)onthecellwallofbacteria&fungiinacalcium-dependentreaction

activating

complementsystem

lysis,

opsonization

promoting

phagocytosis&pathogenclearanceLipidtransfermoleculebindstomonomericLPSandtohigh-affinityLPSreceptornamedCD14and,onmacrophage,neutrophils,DCsLBP+bactericidalpermeabilityincreasingprotein(BPI)bindsLPSonbacteriaandthentoCD14,ahighaffinityLPSreceptor.SeelaterTLR4LPS-bindingprotein(LBP)Extracellularfactor(LPS)carriedbyLBPtoCD14whereitbindstoTLR4andthenMD2bindsSimplifiedVersion模式识别受体（PatternRecognitionReceptors，

PRRs）固有免疫细胞表面、内体、溶酶体、细胞质中、可识别一种或多种PAMPs或DAMPs的识别分子。PRR甘露聚糖凝集素（MBL）C反应蛋白（CRP）血清淀粉样蛋白(SAP)脂多糖结合蛋白（LBP）可溶性：体液和血液细胞吞噬型：细胞膜甘露糖受体（MR）清道夫受体（SR）补体受体（CR）Fc受体（FcR）甲酰甲硫氨酰肽受体（fMLPR）信号转导型细胞膜内体、溶酶体细胞质TLR1、2、4、5、6、10、11、12、13TLR3、7、8、9NLRs、RLRs、ALRsMANNOSERECEPTORThemannosereceptor(MR)isa175kDatypeImembranemoleculeexpressedinthemousebymosttissuemacrophagesandlymphaticandhepaticendothelia. GlycoproteinPRRsrecognizeLPSandlipoteichoicacidIntactG-andG+bacteriaDamagedhostcellsandtissuesApoptoticandsenescentcellsmodifiedlow-densitylipoproteinsSixclassesScavengerreceptors甲酰甲硫氨酰肽受体（fMLPR）StaphylococcalProteinAInhibitsPhagocytosisbyBlockingFc果蝇的Toll受体胞浆的功能域与IL-1受体很相像(Toll/IL-1receptor(TIR)domain)，显然具有重要的免疫功能。Toll突变后果蝇很容易受霉菌感染。Cell86:973-83.Toll-likereceptor(TLR)JulieA.Hoffmann,Ph.D.Strasbourg,FranceIn1996,Hoffmann’sgroupTollfunctionsasaPRRinDrosophilaToll-LikeReceptors(TLRs)Totalof13TLRshavebeenidentifiedinmammalsHuman(TLRs1-10)Mouse(TLRs1-9,11-13)IngeneralTLRsrecognizeconstituentsofmicrobialcellwallsorpathogen-specificnucleicacidsthatareessentialtotheintegrity,functionorreplicationofmicrobes/virusesthatcannotreadilybemodified.Toll-LikeReceptors(TLRs)TherearesixmajorcladesofTLRs,eachrecognizingageneralclassofmolecularpatterns.MoleculartreeofthevertebrateToll-likereceptors(TLRs).TheevolutionofvertebrateToll-likereceptorsToll-LikeReceptors(TLRs):BasicArchitectureEDTMCYTLeaderPeptideLeucineRichRepeats(LRR);BINDINGToll/interleukin1receptor(TIR)domain;SIGNALINGStructuralorganizationofhumanTLRs.ENDOTOXIN-StructuralcomponentoftheouterleafletoftheoutermembraneofGramnegativebacteria-ContainstheO-antigenicpolysaccharidedeterminant.-Presenceofpolysaccharideandlipidcomponents.-AlsotermedLipopolysaccharide(LPS).-ThelipidcomponentorLipidA(glycophospholipid)isresponsibleforthebiologicalactivitiesofLPS(toxicity/fever)LPS(endotoxin)OutermembranePeptidoglycanInnermembraneO-specificchainCoreRegionLipidALPSthetoxiccenterofLPS

TheBasicArchitectureofLPS

StructureofLipidALipidAisaglycophospholipidwithphosphorylatedD-glucosaminesLBPandCD14thefirstEndotoxinReceptorsLPS-BindingProtein(LBP) -PlasmaproteinproducedintheLiver. -DeliversLPSfromtheserumtomacrophages. -EnhancesthesensitivityofmacrophagesforLPS.CD14 -GPI-linkedmembraneprotein(macrophages). -Alsoexistsinasolubleformrecruitedtoendothelialcells. -WorkswithLBPtobringLPStothecellsurface.NolinkbetweenLBP/CD14andintracellularsignaling.However,LPSdetectionbyimmunecellsresultsinanintracellularsignalingresponseandproductionofTNF.DeliveryofLPStoTLR4bylipidtransferproteinsBiologicalActivitiesLBP-CD14Co-receptor(s)MD-2TLR4TLRsignalingpathwaysThegenerationofinflammatorycytokinesandchemokinesThegenerationofantimicrobialpeptidesInitiationSignal-inducedassemblyofpathwaycomponents/involvementofanadaptormoleculeProteinkinase-mediatedphosphorylationInitiationofanenzymecascade:MAPkinasepathway(inmanycelltypes)NFBpathway(apowerfultranscriptionalfactor)generatecytokines,adhesionmolecules&othereffectors

affectthecellcycleorcellulardifferentiationConservedpathwaysininnateimmunityinDrosophilaandmammals.OtherBacterial‘Poisons’LipopeptidesPeptidoglycanLipoteichoicAcidUnmethylatedDNAOligomersTLR4LigandsLipopolysaccharidestwogradesofpurity:

-StandardLPS:LPS-EBandLPS-EKarestandardpreparationsoflipopolysaccharide.Theyareextractedbyaphenol-watermixture.LPS-EBandLPS-EKcontainotherbacterialcomponents,suchaslipopeptides,andthereforestimulatebothTLR4andTLR2.-Ultra-pureLPS:UltrapureLPS-EBareextractedbysuccessiveenzymatichydrolysisstepsandpurifiedbythephenol-TEA-DOCextractionTLR5LigandsFlagellinTLR6/TLR2Ligand-MALP-2TLR2Ligands-Heat-KilledBacteria

-Lipoglycans

-Lipopolysaccharide

-LipoteichoicAcids

-Peptidoglycans

-SyntheticLipoproteins

-ZymosanTLR3LigandsPoly(I:C)asyntheticanalogofdouble-strandedRNA(dsRNA)Poly(A:U)TLR9Agonists

•StimulatoryODNs

-CpGODNs

-ControlODNs

-LabeledODNs

•E.coliDNA

-E.coliDNAef

-E.colissDNA

TLR7Ligands(human&mouseTLR7)

-CL264:Adenineanalog

-

Gardiquimod™:imidazoquinolinecompound

-Gardiquimod™-

Imiquimod:imidazoquinolinecompound

-ImiquimodVacciGradeNEW-

Loxoribine:guanosineanalogueTLR8Ligands(humanTLR8&mouseTLR7)

-

Single-strandedRNAs

-

E.coliRNATLR7/8Ligands(human,mouseTLR7&humanTLR8)

-CL075:thiazoloquinolinecompound

-CL097:water-solubleR848,imidazoquinolinecompound

-Poly(dT):thymidinehomopolymerphosphorothioateODN

-R848:ImidazoquinolinecompoundMyD88-DependentandindependentSignalingTLR4cansignalusingbothpathwaysTRAM=TRIF-relatedadaptormoleculeTIRAP=Toll-interleukin1receptor(TIR)domain-containingproteinTLR4signalinginducesthetranscriptionofbothproinflammatorycytokinesandIFN-thatarerequiredforanti-bacterialandanti-viralimmuneresponses,respectively.CooperationofToll-likereceptorsignalsininnateimmunedefenseNegativeregulationofToll-likereceptor4(TLR4)signalingThenegativeregulatorsofTLRsignalingpathwaysNLRsarecytoplasmicbacterialsensorsthatactivateinflammasomesIL-1familymembersCommonsourcesofinterleukin-1(IL-1)familycytokinesICE:IL-1-convertingenzyme,caspase1Pro-IL-18:24kDaandmatureIL-18:18kDaPro-IL-1beta:33kDaandmatureIL-1beta:17kDapro-caspase-1:45-kDamaturecaspase-1:20KD10KDInflammasomesaremolecularplatformsactivateduponcellularinfectionorstressthattriggerthematurationofproinflammatorycytokinessuchasIL-1bandIL-18toengageinnateimmunedefenses.THEINFLAMMASOMESActivatorsoftheinflammasome.TheHumanNLRFamilyMembers

HumanandMouseNLRFamilyMembersDOMAINORGANIZATIONOFREPRESENTATIVENLRsTheNLRsfamilyiscomposedof>20members.Theycontain:aC-terminalleucine-richrepeat domain,LRRacentralnucleotide-bindingdomain, NACHT

anN-terminalprotein-proteininteractiondomaincomposedof

CARD(caspaseactivationand recruitmentdomain)

Pyrindomain

orBir(baculovirusinhibitorofapoptosisrepeat)domainSchroderandTschoppCell2010NLRP1,NLRP3,IPAF,andAIM2Inflammasomes

Whatisaninflammasomeandwhatdoesitdo?ActivationmodelsandcompositionofNLRP3inflammasomeSchroderandTschoppCell2010PAMPs:MDP,LPS,viral/bacterialRNA,aerolysin,etc.DAMPs:ATP,hyaluronan,uricacid,amyloid-bpeptide,ROS,silica,asbestos,UVB,etc.

ThechannelmodelofNLRP3inflammasomeactivation.ProposedpathwaysforNLRP3activation.AroleformitochondriainNLRP3inflammasomeactivationPathwaysforactivationoftheinflammasomebyPAMPsandDAMPsTheAIM2inflammasomeTheroleoftheinflammasomesinhumandiseaseTheinfluenceofinflammasomeactivationonadaptiveimmunityRLRs等病毒识别受体HostResponsetoVirusInfectionBarriersAdaptiveResponseInnateResponseVirusRoutesofinfectionforVirusesRouteofentryModeofTransmissionPathogenDiseaseMucosalsurfacesAirwayInhaleddropletsInfluenzavirusHantavirusInfluenzaHemorrhagicFeverGastrointestinaltractContaminatedwaterorfoodPoliovirusRotavirusPolioDiarrheaReproductivetractPhysicalcontactPapillomavirusHSV-2CervicalCancer

GenitalherpesExternalepitheliaSkinsurfaceDirectcontactPapillomavirusWartsInstrumentationI.V.InjectionTransfusionTattooingHIVHBVHCVAIDSHepatitisInsectBitesMosquitobites(Aedesaegypti)FlavivirusTogavirusYellowFeverEncephalitisViralNucleicAcidsdoublestranded(ds)RNAsinglestranded(ss)RNADNA(hypomethylatedorCpGrich)“Some”ViralProteins--uncommone.g.glycoproteinsofHerpesvirusandRespiratorySyncytialVirusViralPathogen-AssociatedMolecularPatterns

(PAMP)

ProductionofType1()interferonsProductionofinflammatorymediators:cytokinesandchemokinesMechanism:InterferonResponseFactor3/7(IRF3/7)activationNFBactivationTranslocationfromcytoplasmtonucleusEngagementofViralPAMPReceptorson/inCellscellsurfaceandendosomallocalizationcellcytoplasmlocalizationextracellularspacelocalizationViralPAMPReceptors=anti-viralsensors=anti-viral

PRR

ViralPAMPReceptors(anti-viralsensors)

TLR--TollLikeReceptors:cellsurface&/orendosome

EndosomalTLRTLR-9CpGDNA(

methylatedviralDNA)TLR-7/8ssRNATLR-3 dsRNA [TLR9and7aremajortriggersofType1IFNproduction--particularlybyplasmacytoidDC]:CellsurfaceTLRTLR-2andTLR-4:viralglycoproteins,HCV,RSVThethirdmemberoftheRLRfamily,LGP2,lacksanyCARDsandwasoriginallyidentifiedasanegativeregulatorofRLRsignaling.Nevertheless,LGP2anditsATPaseactivityweredispensablefortheresponsestosyntheticRNAligandsforMDA5andRIG-I.LGP2facilitatesviralRNArecognitionbyRIG-IandMDA5throughitsA