肿瘤免疫课件071103

immunology肿瘤免疫Cancerandtheimmunesystem邱双健复旦大学中山医院肝癌研究所、肝外科1901年以来，免疫学领域研究中有19项获得Nobel奖例如:人类血型的发现(1930)

移植免疫(1991)immunologyTheimmunesystemImmunesystem解剖屏障(皮肤、粘膜等)

生理屏障(温度,pH)

吞噬屏障(细胞吞噬外来分子)

炎症屏障(红、肿、热、痛)

抗原特异性多样性

免疫记忆自身/非自身抗原识别

天然免疫

获得性免疫

immunologyTcells两种类型:HelperTcells(Th):activatesothercellsCytotoxicTcells(Tc):cankillothercellsT细胞识别抗原需有MHC分子的参与immunologyPresentationofantigenstoTcellsProteins(peptides)frominsidethecellarepresentedbyMHCImoleculestoTccells.Proteins(peptides)fromtheoutsideofcellsarepresentedbyMHCIImoleculestoThcells.MHCIonalmostallcellsMHCIIonspecializedantigen-presentingcellsimmunologyT细胞的抗肿瘤作用双重激活信号第一信号：APC细胞的递呈及TCR复合物的形成第二信号：APC共刺激因子（B7、粘附分子等）参与第一信号中，T细胞活化受MHC分子限制-MHC-Ⅰ类分子递呈内源性抗原，激活CD8+T细胞-MHC-Ⅱ类分子递呈外源性抗原，激活CD4+T细胞immunologyT细胞的抗肿瘤作用活化的CD8+T细胞（CTL）是主要的效应细胞CTL分泌释放多种细胞因子及酶表达FasL，于Fas结合，诱导肿瘤细胞凋亡活化的CD4+T细胞通过分泌细胞因子来激活和增强CTL、NK、巨噬细胞的杀瘤效应少数CD4+T细胞也具有直接杀伤肿瘤细胞作用

immunologyThTc病毒感染细胞、肿瘤细胞抗原递呈细胞MHCII+peptideMHCI+peptideimmunologyNK细胞的抗肿瘤作用NK细胞不需致敏即可杀伤多种肿瘤细胞及病毒感染细胞对肿瘤细胞的杀伤不要表现为直接溶解细胞和分泌细胞因子两方面一些肿瘤细胞对NK细胞杀伤的敏感性与存在于肿瘤的MHC分子的变化有关immunology巨噬细胞的抗肿瘤作用作为效应细胞通过非特异性吞噬作用杀伤细胞通过分泌多种细胞毒效应分子直接或间接抑瘤或杀瘤：No、IL-1、TNF、IFN等作为抗原递呈细胞，激发特异性T细胞免疫immunologyNKT细胞的抗肿瘤作用既表达T细胞的表型标记（CD3、TCR)又表达NK细胞的表型标记（CD56)NKT细胞识别的是类似于Ⅰ类MHC的单态性抗原近年的研究表明NKT细胞是肿瘤免疫的执行者，在机体免疫自稳和肿瘤免疫方面有重要意义immunology树突状细胞的作用很少量DC可以激发强大的T细胞反应可以激活静息T细胞，这种作用与DC-SIGN分子有关体内可以致敏CD4+T辅助细胞和CD8+T杀伤细胞immunologyCancerThesecondrankingcauseofdeathafterheartdiseaseintheWesternworld.mostorgansandtissuesinanorganismareinbalance(deathandrenewal)

cancercellshavenocontrolingrowthmechanisms,canexpandtoalargesizeproducingatumor

immunologyTumorgrowthandmetastasisimmunology免疫缺陷与癌症的发生密切相关原发性免疫缺陷淋巴瘤Burkitt’s

淋巴瘤

疟疾

继发性性免疫缺陷淋巴瘤,宫颈癌,肝癌,皮肤癌,Kaposi’s肉瘤.

自身免疫淋巴瘤肿瘤免疫缺陷的病因肿瘤刺激机体的免疫应答动物可对肿瘤产生免疫免疫力可转移给其他动物在患有一些恶性肿瘤的人体，已经能检测到肿瘤特异性抗体和细胞抗肿瘤免疫机制Immunesystem补体的溶细胞效应CDC效应抗体依赖的细胞介导的细胞毒效应

ADCC效应抗体的免疫调理作用抗体对肿瘤细胞的封闭作用抗体干扰肿瘤细胞的黏附作用

T细胞的抗肿瘤作用

主要是细胞毒性T淋巴细胞NK细胞的抗肿瘤作用巨噬细胞的抗肿瘤作用

作为效应细胞通过非特异性吞噬作用通过分泌多种细胞毒效应分子起作用作为抗原递呈细胞NKT细胞同时有T、NK细胞表型树突状细胞

体液免疫机制

细胞免疫机制

immunology肿瘤抗原两种类型:肿瘤相关抗原（TAA、TATAs)

正常组织也有不同程度表达肿瘤特异性抗原（TSA、TSTAs)

仅仅表达于肿瘤细胞immunology肿瘤抗原的现代分类immunology抗原类型人类肿瘤抗原举例癌基因、肿瘤抑制基因Ras突变产物、Her-2/neu产物、p53突变产物等与癌变无关的基因突变产物黑色素瘤的能为CTL识别的突变蛋白大多数正常组织中静止的基因产物黑色素瘤的MAGE、BAGE、GAGE抗原正常情况下主要在睾丸和胎盘中表达过度表达的基因产物如黑色素瘤等的酪氨酸激酶、gp100、MART肿瘤相关病原体病毒产物HP、人乳头状病毒E6、E7蛋白、SV40T抗原等胚胎性抗原CEA、AFP糖脂和糖蛋白GM2、GD2（黑色素瘤）来自起源组织的分化抗原PSA、淋巴细胞标记（CD10、CD20以及Ig独特型）寻找肿瘤特异性抗原肿瘤细胞cDNA文库转染表达相应HLA分子的靶细胞肿瘤细胞表面分子或纯化的HLA分子洗脱肽反向免疫法寻找肿瘤细胞特异的高表达分子重组cDNA表达文库的血清学鉴定技术等蛋白组学高通量筛选技术immunology肿瘤免疫逃逸机制被动机制肿瘤细胞表面MHC分子表达及抗原处理异常缺乏抗原性表位缺乏共刺激因子Fas/FasL系统的异常表达immunology肿瘤免疫逃逸机制主动机制肿瘤组织产生免疫抑制因子T细胞的异常

消除肿瘤特异性T细胞诱导免疫抑制功能T细胞产生引起T细胞的耐受引起TAA特异性的T细胞无能抑制DC的分化、成熟TCR及IL-2受体通路异常

CD4+/CD8+倒置和Th1/Th2飘移肿瘤抗原被封闭其他方面：膜结合补体调节蛋白、黏附分子缺失、癌基因过度表达等T调节细胞（CD4、CD25共表达T细胞）的增加immunology肿瘤的免疫治疗非特异性卡介苗,短小棒状杆菌,左旋咪唑等.杀伤的肿瘤细胞,纯化或基因重组的肿瘤抗原特异性主动免疫治疗LAK细胞,细胞因子非特异性抗体单独或与其他试剂结合,激活的T淋巴细胞特异性被动免疫治疗肿瘤特异性抗原的应用制备单克隆抗体用于诊断用于治疗在体内刺激特异性免疫应答特异性主动治疗特异性被动治疗肿瘤特异性抗原制备的单克隆抗体的应用单克隆抗体可作为诊断的工具非特异性免疫治疗激活巨噬细胞和NK细胞IFN-α,IFN-β,IFN-γ,IL-2,TNF-α细胞因子产生干扰素吡喃,多聚I:C合成分子激活巨噬细胞和

NK细胞(通过细胞因子)卡介苗,短小棒状杆菌,胞壁酰二肽细菌产品肿瘤免疫治疗进展肿瘤疫苗研究细胞因子基因修饰的疫苗转染共刺激因子的肿瘤疫苗抗原特异性疫苗细胞因子在肿瘤免疫治疗中的应用增强抗肿瘤潜能的研究DC疫苗在肿瘤免疫治疗中的应用肿瘤抗原修饰的DC疫苗其它细胞因子等修饰的DC疫苗immunology肿瘤疫苗的研究早期试图用死亡的肿瘤细胞或裂解产物来激发特异的免疫反应目前主要集中在基因工程疫苗的研究－细胞因子基因修饰的瘤苗－转染共刺激因子的瘤苗immunology抗原特异性疫苗多肽疫苗几乎都是MHCⅠ分子限制性的抗原肽，有成功报道，但有可以导致免疫耐受重组病毒免疫常用痘病毒或腺病毒载体，表达相关的肿瘤抗原重组细菌疫苗多用沙门杆菌、卡介苗等核酸疫苗使用编码肿瘤抗原的裸DNA作疫苗immunologyDC疫苗的研究肿瘤抗原修饰的DC疫苗其他因子修饰的DC疫苗：细胞因子、趋化因子等DC疫苗已进入临床实验阶段，初步的研究表明：DC免疫不会引起明显的急性毒性反应；不会产生有明显临床意义的自身免疫性疾病immunology增加MHC-I分子的表达，可能为抗肿瘤机制缓解毛细胞白血病,对癌症有弱的效应IFN-α,-βIFN-β增加MHC-I分子的表达,激活Tc

和NK细胞缓解卵巢癌IL-2使T细胞活化和增殖,激活NK细胞缓解肾细胞癌和黑色素瘤TNF-α

激活巨噬细胞和淋巴细胞减少恶性腹水细胞因子用于免疫治疗增强抗肿瘤免疫潜能的研究重视TNF家族的免疫调节作用热休克蛋白作为抗原载体重视CD4+T细胞的抗肿瘤作用重视DC疫苗的体内效能发挥重视各种方法的联合应用immunology重视与其他治疗手段综合、序贯应用将最新进展转化为临床实践可操作的手段任何免疫治疗都必须与其他治疗手段如手术、放疗、化疗相结合精心优化治疗方案，设计行之有效的肝癌综合治疗策略，降低肝癌转移复发问题与展望肿瘤细胞免疫逃避机制未充分了解重视肿瘤微环境对T细胞的抑制作用实验研究向临床实践的转换immunologyCancerDespiteImmunosurveillance:

ImmunoselectionandImunosubversionShuang-JianQiu,MD.,PhD.ZitvogelL,etal.NatureRevImmunol2006;6:715-727OutlineIntroductionCancerimmunosurveillanceIntrinsicandextrinsticbarrierstocarcinogenesisThesixcell-intrinsichallmarksofcancerTheseventhhallmarkofcancerConcludingremarksSevenhallmarkersofcancerCell-intrinsicProvidetheirowngrowthsignals;Ignoregrowth-inhibitorysignals;Avoidcelldeath;Replicatewithoutlimits;Sustainangiogenesis;Invadetissue;Cell-extrinsicAvoidimmunosurveillance.Theimmunesystemcanrecognizeanddestroyprecursorsofcancerbeforeclincallyapparent.InitiallyproposedbyPaulEhlrichacenturyagoFormallyintroducedbyBurnetandThomasinthe1950'sCriticisedbyPrehnandothers”Immunostimulationtheory”considereddeadBy1978Since1994keyfindingsrekindleinterestincancerimmunosurveillanceAbroaderterm,immunoediting,emergedImmunosurveillance–historicalperspectivesCD8+

CTLsrecognizeandkillstromaland/ortumorcellssecretetheanti-angiogeniccytokineIFNγDefinedcancerimmunosurveillance(a)convertM1macrophagesintoM2macrophagesreleaseIFNγ,whichinhibitsangiogenesisproduceIL-4andblockneo-angiogenesisCD4+TcellsDefinedcancerimmunosurveillance(b)Definedcancerimmunosurveillance(c)killtumorcellsinTRAILandperforin-dependentmanneramajorsourceofIFNγcross-presenttumorantigenstoTcellsIFN-producingkillerdendriticcells(IKDCs)

activatedbyDCindiversemechanismsNK

cellsDefinedcancerimmunosurveillance(d)Definedcancerimmunosurveillance(e)recognizeglycolipidsboundtoCD1datthesurfaceofAPCsecreteIFNγinaCD40-andIL-12-dependentmannerlysetumorcellsthoughTRAILorperforin-dependentpathwayNKT

cellsDefinedcancerimmunosurveillance(e)Definedcancerimmunosurveillance(f)TumorcellsorAPCs

activateγδTcellsproducecytokineskilltumorcellsinanNKG2D-dependentmannerγδTcellsDefinedcancerimmunosurveillance(f)Definedcancerimmunosurveillance(g)inducingcomplement-mediatedlysis

ADCbyNKcellsandmacrophagesTumor-specificAbsDefinedcancerimmunosurveillance(g)AvoidanceofimmunosurveillanceImmunoselectionOutgrowthof

poorly/non-immunogenictumorcellvariants;Alsoknownasimmunoediting;ImmunosubversionSuppressimmuneresponsesactively.Cell-extrinsicmechanismsCooperatewiththesixcell-intrinsiccharacteristicsmechanistically.ImmunoselectionGenomicinstabilitygivesrisetogeneticdiversityintumors.NaturalselectionoftumorvariantsoccursbydifferentialpropagationoftumorsubclonesThisalsoappliestotumorgrowthaftereffectiveimmunotherapythreeEsofcancerimmunoeditingCancerimmunoeditingencompassesthreeprocess.(a)Eliminationcorrespondstoimmunosurveillance.(b)Equilibriumrepresentstheprocessbywhichtheimmunesystemiterativelyselectsand/orpromotesthegenerationoftumorcellvariantswithincreasingcapacitiestosurviveimmuneattack.(c)Escapeistheprocesswhereintheimmunologicallysculptedtumorexpandsinanuncontrolledmannerintheimmunocompetenthost.Crosstalkofcancer-intrinsicfactorsandhostimmunesysteminmulti-stepcarcinogenesisSixcell-intrinsichallmarkersofcancerImmunesystemIntrinsicandextrinsicbarrierstocarcinogenesisDNAdamageIntrinsic

activatetheATM–CHK1–p53pathway;leadtocell-cyclearrestorapoptosis.Extrinsic(immune-mediated)ActivatetheexpressionofligandsforNKG2Dbytumorcells;RenderthetumorcellsimmunogenicandsusceptibletokillingbyNKG2D-expressingNK,NKT,Tcells.TypeIIFNandp53TypeIIFNhaveanimportantroleinimmunosurveillance;TypeIIFN

upregulatesthep53-mediatedresponseoftumorcells;Lossofp53increasessusceptibilitytotype-I-IFN-induced,TRAIL-dependentapoptosis.IntrinsicandextrinsicbarrierstocarcinogenesisSelf-sufficiencyingrowthsignalsIL-4,IL-6,IL-10IntrinsicAutocrinesignalsbytumorcells;Extrinsic(immune-mediated)PromotedifferentiationintoTH2cells;Inhibitcrosstalkbetweeninnateandadaptiveimmuneresponse;StimulateaB7-H4-expressingT-cellsuppressingmacrophagepopulation.Impactof

CancerintrinsicCharacteristicsonIS(a)TGF-βIntrinsicTumorsdisablescomponentsofTGF-β-mediatedsignalpathway;TGF-β

favourstheepithelialtomesenchymaltransitionoftumorcells.Extrinsic(immune-mediated)Inhibitsantigenpresentation;InhibitsTcellproliferation;InhibitsNKcellcytotoxity;ActivateregulatoryTcell.InsensitivitytoantigrowthsignalsImpactof

CancerintrinsicCharacteristicsonIS(b)BCL-2,MUC1,SurvivinIntrinsicUpregulatedanti-apoptoticproteins.Extrinsic(immune-mediated)Theyaretumorassociatedorspecificantigens;MUC1inhibitsDCdifferentiation,maturationandinducesaregulatoryphenotype;SurvivinpromotesTcellapoptosisthroughinducingCD95Lexpressingbytumorcells;EvasionofapoptosisImpactof

CancerintrinsicCharacteristicsonIS(c)APAF1(apoptotic-protease-activatingfactor1),IAPs(inhibitor-of-apoptosisproteins)IntrinsicInactivationofAPAF1Overexpress

IAPsExtrinsic(immune-mediated)LackofcaspaseactivationaffecttheimmunogenecityofcelldeathReducetheimmunogenecityofapoptosisEvasionofapoptosisImpactof

CancerintrinsicCharacteristicsonIS(d)Mutatedp53,TERT(Telomerasereversetranscriptase)IntrinsicMaintainoftheendsofchromosomes;Ensurereplicativepotential.Extrinsic(immune-mediated)Tumorantigens.LimitlessreplicativepotentialImpactof

CancerintrinsicCharacteristicsonIS(d)VEGF,COX2IntrinsicHigherexpressionbytumorcells;Mediateaproangiogenic

swithExtrinsic(immune-mediated)InhibitT-cellactivationandcytotoxity;PromotesTH2-cellresponseInhibitNF-kBactivationinDCs.SustainedagiogenesisImpactof

CancerintrinsicCharacteristicsonIS(e)MMPs(matrixmetalloproteinase)IntrinsicSecretedbytumorcellsandstromalcells;Promotetumor-inducedangiogenesis,invasionandestablishmentofmetastaticfoci.Extrinsic(immune-mediated)InducecleavageofCD25;ActivateTGF-β;PromotesheddingofICAM1andICAM2.TissueinvasionandmetastasisImpactof

CancerintrinsicCharacteristicsonIS(f)NECL2(nectinlike2)atumor-suppressorgeneencodedprotein;Mediatesepithelialcelljunctionsthroughnectinlikeproteins.IntrinsicInactivatedintumorcells;Allowsthedisorganizationofepithelial-celllayers.productionExtrinsic(immune-mediated)InhibitsNKcellcytotoxity;InhibitsCD8+TcellIFNγ

production;TissueinvasionandmetastasisImpactof

CancerintrinsicCharacteristicsonIS(f)TheseventhhallmarkofcancerTumorsevadeorparalyseimmunesystemTumorvariantsloseantigen-processingmachinery,tumorantigensandsensitivitytoimmuneeffectorssuchasinterferons(IFNs)Mech.tumorescapefromimmunesystem(a)Tumor-derivedfactorsrecruitmyeloidsuppressorcells(MSCs)andpreventtheirdifferentiationintomatureDCsMSCsinhibittumour-specificTcellsthrougharginase-1(ARG1)ornitric-oxidesynthase2(NOS2)Mech.tumorescapefromimmunesystem(b)InteractionbetweenCD80orCD86andCTLA4inducesIFNγandimmunosuppressivefactorIDObyDCsThisresultsinareductionintheamountoftryptophanandinthegenerationofkynurenines,whichkillTcells.Mech.tumorescapefromimmunesystem(c)pDCsactivatedbyIL-3andCD40Lpromotethediffer.intoTH2cellsandIL-10-producingCD8+TregThisstateofanergy(withrespecttotumour-celllysis)ismediatedbyIL-10,directlyorindirectlyMech.tumorescapefromimmunesystem(d)RepetitivestimulationofnaiveTcellswithimmatureDCsresultsinT-cellanergy,togetherwithIL-10productionandIL-10-independent,cell-contact-dependentregulatoryactivityMech.tumorescapefromim