肿瘤微环境PPT参考幻灯片

肿瘤免疫中的正性和负性免疫成份,居颂光,1,细胞出版社以增刊方式向世界专门介绍中国免疫学研究,国际著名的细胞出版社（CellPress）近日推出新一期聚焦中国(SpotlightonChina)，介绍近年来中国免疫学基础及临床研究的飞速发展，其电子版在细胞出版社及其子刊的网站上均能免费阅读或下载,2,KnutsonK,etal.CancerImmunolImmunother2005,54:721728,APCTHTregCytoxicTcellsOthers,IFNIL-2IL-12GranzymePerforinAbsothers,Tumormicroenvironment:,3,肿瘤研究肿瘤细胞研究,一个世纪的观念：肿瘤是局部组织/器官细胞异常生长形成的疾病肿瘤是个系统性疾病肿瘤是一种组织（间质、血管、微环境），不是一堆肿瘤细胞免疫功能失控，肿瘤患者的免疫系统不是简单的低下：保护肿瘤生长的免疫细胞：功能增强抑制肿瘤细胞的免疫细胞：功能低下,4,肿瘤的免疫编辑,免疫成分在肿瘤发生过程中的作用,5,DunnG,etal.Natureimmunology.2002.3(11):991,Eliminationcorrespondstoimmunosurveilliance,Thehostimmunesystemandanytumorvariantthathassurvivedintheeliminationprocessenterintothedynamicequilibrium:*lymphocytesandIFN-exertpotentselectionpressure.*Atumorbediscontainingmanygeneticallyunstableandrapidlymutatingtumorcells.,Survivaltumorvariantsthathaveacquiredinsensitivitytoimmunologicdetectionand/oreliminationbegintoexpandinuncontrolmanner.,DunnG,etal.Natureimmunology.2002.3(11):991,6,Solidtumorsreachacertainsize:Growinvasively,requireanenhancedbloodsupply.Inflammatorysignalsleadingtorecruitmentofcellsofinnateimmunesystem:macrophages,DCs,-T,NKandNKT.ProduceIFN-,7,TheinitialIFN-startsacascadeofimmuneresponse:1.Inductionofchemokines:CXCL10,CXCL9,CXCL11.Blockneovascularizationoftumor,recruitmacrophages,DCs,-T,NKandotherimmunecells.2.Antiproliferationofdevelopingtumor.3.TheactivationofcytocidalactivityinmacrophagesandNKcellsenteringthetumor.DeadtumorcellsortumorcellsdebrisareingestedbyDCandaretraffickedtothedraininglymphnode.,8,1.TumorgrowthiskeptincheckbythecytocidalNKandmocrophages.2.CD4+TandCD8+Tcellsthatarespecificfortumorantigensaredevelopingindraininglymphnode.,9,TumorspecificCD4+TandCD8+Tcellshometotumorsitealongachemokinegradient.,10,DunnG,etal.Natureimmunology.2002.3(11):991,Eliminationcorrespondstoimmunosurveilliance,Thehostimmunesystemandanytumorvariantthathassurvivedintheeliminationprocessenterintothedynamicequilibrium:*lymphocytesandIFN-exertpotentselectionpressure.*Atumorbediscontainingmanygeneticallyunstableandrapidlymutatingtumorcells.,Survivaltumorvariantsthathaveacquiredinsensitivitytoimmunologicdetectionand/oreliminationbegintoexpandinuncontrolmanner.,11,肿瘤研究肿瘤细胞研究,一个世纪的观念：肿瘤是局部组织/器官细胞异常生长形成的疾病肿瘤是个系统性疾病肿瘤是一种组织（间质、血管、微环境），不是一堆肿瘤细胞免疫功能失控，肿瘤患者的免疫系统不是简单的低下：保护肿瘤生长的免疫细胞：功能增强抑制肿瘤细胞的免疫细胞：功能低下,12,肿瘤抗原,OliveraJ.Finn,Ph.D.,NEnglJMed2008;358:2704-15,OliveraJ.Finn,Ph.D.,NEnglJMed2008;358:2704-15,13,肿瘤抗原的加工提呈,OliveraJ.Finn,Ph.D.,NEnglJMed2008;358:2704-15,14,肿瘤干细胞,15,KnutsonK,etal.CancerImmunolImmunother2005,54:721728,APCTHTregCytoxicTcellsOthers,IFNIL-2IL-12GranzymePerforinAbsothers,Tumormicroenvironment:,16,NEnglJMed2006;355:1253-1261.,17,Cancerstemcell,NatureReviewsCancerdoi:10.1038/nrc1232,TumorHeterogeneityandCancerStemCells,18,免疫成分在肿瘤微环境中的失衡,19,Nature.2005,5:263,20,21,22,23,24,25,肿瘤微环境中的免疫细胞,26,TH17cellsintumourimmunityandImmunotherapy.NATuREREVIEWS|Immunology.doi:10.1038/nri2742,DifferentiationofhelperTcellsubsets,27,28,TH17cellsandantitumourimmunity,29,30,肿瘤微环境中的抑制性成分Treg细胞,31,Treg细胞定义,1995由Sakaguchi等发现了机体重要的负性免疫调节成分调节性CD4+CD25+T细胞（regulatoryTcell,Treg）。随后的研究根据细胞内标记分子Foxp3和细胞膜分子CD127表达与否，将经典的Treg细胞定义为CD4+CD25+Foxp3+CD127low/negT细胞。通过细胞间接触依赖机制发挥作用或分泌IL-10和TGF-等细胞因子，Treg细胞抑制CD4+25-T细胞、CD8+T细胞和树突状细胞等其他免疫细胞的活性和功能。,32,大量的研究报道表明:CD4+CD25+Treg细胞是抑制机体抗肿瘤免疫应答的关键成分之一，例如：Woo等发现在非小细胞肺癌和卵巢癌患者的肿瘤浸润淋巴细胞中Treg的含量明显增多；在胃肠道肿瘤患者中CD4+CD25-T细胞明显减少而Treg细胞数量增多，而且Treg细胞数量的增高与较差的预后和较低的生存率高度相关；Curiel等报道，在卵巢癌中Treg细胞能抑制T细胞介导的特异性抗肿瘤免疫应答，并且有利于肿瘤的生长；因此研究者希望通过减少Treg细胞的数量或降低其活性来开辟治疗肿瘤免疫治疗的新途径。例如，以Foxp3为靶点清除荷瘤小鼠体内的Treg细胞后肿瘤会消退。,33,Treg细胞的作用机制,34,非小细胞性肺癌（NSCLC）与Treg新亚群,35,Fig1,36,Fig.2,37,Fig3,38,Fig4,39,CD8+T细胞在肿瘤免疫中的作用,肿瘤微环境中的“正性”免疫成分,40,Gadd45-/-miceorHet/WTmicewereinjectedintointradermallyofflankskinwith2.5X104B16cells/mouse.,Tumorsizesweremeasuredevery2days.,Tumorinjectsite,Gadd45bKOmicewerechallengedwithmelanomacells:B16,Freezedownthetissueoftumorsite.,41,Fig4Gadd45-/-miceorWTmicewereinjectedsubcutaneouslywith2.5X104B16cells/mouse,tumorsizesweremeasuredevery2days.Thesedataarefrom3separatedexperiments.,42,5.BothWTandGadd45bKOTcellsareabletoinfiltrateintotumorsites.,43,Gadd45-/-miceorWTmicewereinjectedintradermallyofflankskinwith5X104B16cells/site/20ulPBS,Skinofinjectedsitesoroppositesiteswasresectedandfreezedownimmediately,SpleenCellsAdoptiveTransferandB16inoculation,Tumorinjectsite,Checktumorformationfrominjectedsites,500Rad,WT,KO,500Rad,Donor,Host,44,6.Gadd45isimportantforlymphocytesintumorsurveillance,P0.05,*,45,CD4+TCellsAdoptiveTransferandB16inoculation,500Rad,WTCD4+T,KOCD4+T,500Rad,Donor,Host,WTsplenocytesdepletedCD4+T,WTsplenocytesdepleteCD4+T,+,+,46,7.Gadd45bKOdoesnotaffectfunctionofCD4+Tcellsintumorsurveillance,KO:TransferwithCD4+TcelldepletedspleencellsfromWTmice+CD4+TcellsfromGadd45bKOmice,PBS:OnlytransferPBS,WT:TransferwithCD4+TcelldepletedspleencellsfromWTmice+CD4+TcellsfromGadd45bHetorWTmice,47,CD8+TCellsAdoptiveTransferandB16inoculation,500Rad,WTCD8+T,KOCD8+T,500Rad,Donor,Host,WTsplenocytesdepletedCD8+T,WTsplenocytesdepleteCD8+T,+,+,48,8.Gadd45bKOcausesfunctiondefectofCD8+Tcellsintumorsurveillance,KO:TransferwithCD8+TcelldepletedspleencellsfromWTmice+CD8+TcellsfromGadd45bKOmice,PBS:OnlytransferPBS,WT:TransferwithCD8+TcelldepletedspleencellsfromWTmice+CD8+TcellsfromGadd45bHetorWTmice,P0.05,*,49,Gadd45bisexpressedinCD8+Tcellsduringtheiractivation,Figure1Gadd45bmessageisinducedbyTCRsignalingandcostimulation.NaiveCD8+Tcellswerestimulatedwithplateboundanti-CD3(-CD3)plus-CD28andTh1conditionfor0h,1h,12h,24h,48hand96h.TheGadd45bmessagewasdetectedwithRT-PCR.,50,2.Gadd45isimportantforthefunctionofCD8+Tcells.,IL12/IL18,-CD3,51,52,MAPkinaseprofileactivatedinCD8+Tcells,-CD3(10g/ml),IL-12(10ng/ml)+IL-18(30ng/ml),Figure3Gadd45mediatesp38activationbyTCRsignalingorIL-12plusIL-18inCD8+Tcells.(a)EffectorCD8+Tcellswerestimulated(times,abovelanes)withplate-boundanti-CD3(-CD3;10g/ml)or(b)IL-12(10ng/ml)andIL-18(30ng/ml).Cellextractswereanalyzedbyimmunoblotwithanti-phospho-ERK(-phospho-ERK),-phospho-JNKand-phospho-p38.p-,phosphorylated.,53,Gadd45bKOcausesfunctiondefectoftumorinfiltratingCD8+Tcells,Fig5Gadd45KOcausetumorinfiltratingCD8+Tcellfunctiondefectinvivo.Tumormasswasremovedatday13afterB16F0cellsinoculationanddigestedwithcollagenaseI.Cellswerestimulatedwithphorbol12-myristate13-acetate(50ng/ml)andionomycin(500ng/ml)for6handincubatedforthelast3hwithBrefeldinA.CellsproducingIFN-wasidentifiedwithintracellularcytokinestaining.,54,Type,Density,andLocationofImmuneCellsWithinHumanColorectalTumorsPredictClinicalOutcome,GalonJ,etal.29SEPTEMBER2006VOL313SCIENCE,55,Type,Density,andLocationofImmuneCellsWithinHumanColorectalTumorsPredictClinicalOutcome,GalonJ,etal.29SEPTEMBER2006VOL313SCIENCE,(B)ComparisonofthemeanofimmunecelldensitiesintheCTandIMfrompatientswithtumorrecurrence(blackbars)orwithouttumorrecurrence(whitebars).,56,肿瘤微环境中的Immunecheckpoints,NATUREREVIEWS|CANCER2012doi:10.1038/nrc3239,57,Multipleco-stimulatoryandinhibitoryinteractionsregulateTcellresponses,58,Immunecheckpointsregulatedifferentcomponentsintheevolutionofanimmuneresponse,59,Twogeneralmechanismsofexpressionofimm