细胞过继免疫治疗肿瘤的临床研究进展ppt课件

细胞过继免疫治疗肿瘤的临床研究进展,1,细胞免疫治疗肿瘤的发展,上世纪80年代初，StevenA.Rosenberg,ChiefofSurgeryattheNationalCancerInstitute，NIH，USLAK+IL-222例中：CR1，PR8（41%）LAK、TIL、基因修饰、联合放、化疗,2,国内细胞免疫治疗研究热潮,80年代末90年代初LAK推动了国内细胞免疫治疗研究的热潮。LAK：自体、异体、胎儿脾、胸腺、脐血90年代，我国药品监督管理局制定了相应法规，并公布实施。此后又进行了多次修订。首例申报CIK，北京人民医院（陆道培）DC疫苗，第二军医大学（曹雪涛）上个世纪末CIK、DC-CIK、NK、gdT、TIL、免疫杀手细胞等开展了多种免疫细胞抗肿瘤研究。,3,肿瘤的细胞免疫治疗经过体外培养加工,细胞过继免疫治疗(adoptiveimmunotherapy)LAK、TIL、CIK、DC-CIK、NK、NKT、CTL、gdT自体/异体，基因修饰细胞瘤苗（tumorvaccine）肿瘤细胞树突状细胞自体/异体、基因修饰融合细胞,4,CIKCytokineinducedkiller,PBMCIFN-ganti-CD3,IL-2,IL-1a,5,2009，Germany，Prof.Dr.Schmidt-Wolf建立了CIK临床研究登记网站，目前收集发表的文章39篇，中国人发表26篇(占2/3)。2011，JCancerResClinOncol.“ClinicaltrialsonCIKcells:firstreportoftheinternationalregistryonCIKcells(IRCC)”.PubMedthekeywordsCIKcellsclinicaltrials，evaluated11studies（426人）.并与作者联系核实涉及的杂志：DigestiveandLiverDisease,ChineseJournalofCancer,AnticancerResearch,WorldJournalofGastroenterology,Haematologica,BritishJournalofCancer,BiologyofBlood,andMarrowTransplantation.Insomestudies,abstractswereonlyavailableinChinese.Unfortunately,thesestudiesareoftenpublishedonlyinCernationalstandard,CIK临床研究国际登记网站,6,ClinicaltrialsonCIKcells:firstreportoftheinternationalregistryonCIKcells(IRCC),C.HontschaY.BorckH.ZhouD.MessmerI.G.H.Schmidt-WolfJCancerResClinOncol(2011)137:305310,入组标准排除标准,Table2.IndicationsandcontraindicationsofCIKimmunotherapyin11clinicaltrials,7,Table1Tumorentitywithnumberofrespectivepatients,male-to-femaleratio,andtherapeuticeffects,ClinicaltrialsonCIKcells:firstreportoftheinternationalregistryonCIKcells(IRCC),脊髓发育不良,8,CytokineInducedKillerCellsasPromisingImmunotherapyforSolidTumorsDarioSangiolo,ItalyJournalofCancer2011;2:363-36,CIK治疗实体瘤的9个临床研究报道,9,Fig.1.Disease-freesurvivalcurvesofthethreegroups.KaplanMeierwasemployedtoestimatethedisease-freesurvivalratesofthethreegroups.LogranktestshowsasignificantlyhigherratesintheCIK-Igroup(p=0.001)andtheCIK-IIgroup(p=0.004)thaninthecontrolgroup.NostatisticalsignificancewasfoundbetweentheCIK-IgroupandtheCIK-IIgroup(p=0.345).,Arandomized,controlledtrialofpostoperativeadjuvantcytokine-inducedkillercellsimmunotherapyafterradicalresectionofhepatocellularcarcinoma（127例）HuiDong,QiangLi,JianWang,TiZhang,Da-LuKong,DigestiveandLiverDisease41(2009)3641DepartmentofHepatobiliarySurgery,CancerHospitalofTianjinMedicalUniversity,Tianjin,PeoplesRepublicofChina,无病生存期肝癌切除术后1个月开始治疗，CIK-Ix3次CIK-IIx6次对照,10,Fig.2The1-,3-and5-yeardisease-freesurvivalratesinpatientsreceivingCIKcells(groups1and2)whencomparedtonoCIKcelltreatment(127patients);(trial11).Group1wastreatedwith3coursesofCIKcells,group2wastreatedwith6coursesofCIKcellsandthecontrolgroup(group3)receivednoCIKcelltreatmentforcomparison,1-,3-和5-年无病生存率,Arandomized,controlledtrialofpostoperativeadjuvantcytokine-inducedkillercellsimmunotherapyafterradicalresectionofhepatocellularcarcinoma（127例）,11,Arandomized,controlledtrialofpostoperativeadjuvantcytokine-inducedkillercellsimmunotherapyafterradicalresectionofhepatocellularcarcinoma,1-,3-and5-yearoverallsurvivalrates,生存期,12,Figure1Survivalratesofpatientsaftercytokine-inducedkillercellsimmunotherapypluschemotherapyandchemotherapyalone.CIK:Cytokineinducedkiller.,CIK治疗胃癌频率与疗效IncreasingthefrequencyofCIKcellsadoptiveimmunotherapymaydecreaseriskofdeathingastriccancerpatients(156例)Jing-TingJiang,.Xue-GuangZhang苏州大学WorldJGastroenterol2010December28;16(48):6155-6162,13,WorldJGastroenterol2010December28;16(48):6155-6162Jing-TingJiang,Yue-PingShen,Chang-PingWu,Yi-BeiZhu,Wen-XiangWei,Lu-JunChen,XiaoZheng,JingSun,Bin-FengLu,Xue-GuangZhang苏州大学,IncreasingthefrequencyofCIKcellsadoptiveimmunotherapymaydecreaseriskofdeathingastriccancerpatients,14,Cordblood-derivedcytokine-inducedkillercellsbiotherapycombinedwithsecond-linechemotherapyinthetreatmentofadvancedsolidmalignancies(40例),QiNiu,et.al.,InternationalImmunopharmacology11(2011)449456BeijingGreatWallInternationalCancerCenter,No.309PLAHospital,Beijing,PRChina,中位生存期化疗7.52个月化疗+CIK11.17个月差3.65个月,化疗联合脐血CIK治疗恶性肿瘤,15,Cordblood-derivedcytokine-inducedkillercellsbiotherapycombinedwithsecond-linechemotherapyinthetreatmentofadvancedsolidmalignancies,16,中国肺癌杂志2009,Vol.12,No.9昆明医学院第三附属医院赵光强黄云超叶联华段林灿周永春杨凯云马千里雷玉洁宋鑫黄明杨银山,全身化疗和支气管动脉灌注联合CIK过继免疫治疗期非小细胞肺癌的临床研究（113例）,17,3CR,1PR,6SD，6PD/16CANCERBIOTHERAPYANDRADIOPHARMACEUTICALS25%62.5%Volume25,Number4,2010,ImmunotherapywithCytokine-InducedKillerCellsinMetastaticRenalCellCarcinoma,XiaosanSu，etal.,TheFirstHospitalofKunming,Kunming,PeoplesRepublicofChina.,18,目前对CIK治疗肿瘤的总体评价,相对安全材料、方法不一致评价标准不统一对血液系统来源的恶性肿瘤疗效好对实体瘤的疗效有待确定CIK应用前景较好；异体CIK克服同种异体排斥反应后是较好的治疗手段。,19,PBMCd0,粘附细胞,非粘附细胞,GM-CSF,IL-4,IFN-rAnti-CD3,IL-2,IL-1,Ag,TNF-a,CIK,DC,DC-CIK,Day5-9,DC-CIK,20,共培养的树突状细胞与CIK细胞治疗结肠癌血源性肺转移的实验研究,张嵩王恩忠白春学徐永华中围科学院上海生命科学院生物化学与细胞生物学研究所，旦大学附属中山医院肿瘤2003年第23卷6期,Lovo人结肠癌健康人血,21,共培养的树突状细胞与CIK细胞治疗结肠癌血源性肺转移的实验研究,22,裸鼠接种Lovo人结肠癌1x107ivday3,7,10,P0.05,DC-CIK抑瘤效果优于CIK,23,提高急性白血病患者完全缓解病人的无病生存期陆道培教授领导的研究小组用自体CIK或自体DC-CIK治疗了111例经化疗或（和）自体造血干细胞移植（ASCT）治疗后完全缓解（CR）的急性白血病患者。96例CR患者再接受CIK或DC-CIK治疗后，5年无病生存率（DFS）为66%，明显高于在该医院接受单纯化疗或ASCT治疗的患者。1415例残留的白血病染色体和（或）基因标志均转阴。无一例发生严重不良反应。主要不良反应为畏寒、寒战、发热、疲乏，均在24小时内消失。,24,该研究小组采用自CIK或自体DC-CIK治疗了24例白血病合并丙肝病毒感染的患者。22例中有16例的血HCV-RNA转阴；在接受2疗程治疗的患者中，15例中有9例患者(60%)血中HCV-RNA持续转阴。,使白血病合并丙肝病毒感染的患者HCV-RNA持续转阴,25,Autologouscytokine-inducedkillercelltherapyinclinicaltrialphaseIissafeinpatientswithprimaryhepatocellularcarcinomaMingShi,Fu-ShengWang,DivisionofBiologicalEngineering,InstituteofInfectiousDiseases,302HospitalofPLA,BeijingWorldJGastroenterol2004;10(8):1146-1151,CIK抗乙肝病毒作用,26,Autologouscytokine-inducedkillercelltherapyinclinicaltrialphaseIissafeinpatientswithprimaryhepatocellularcarcinoma,Thegrowthoftumorsinallpatientsbecameslowdown,thetumorvolumewasdecreasedin3patients.,乙肝病毒测定,27,DC+CIK联合化疗治疗晚期非小细胞肺癌的临床疗效评价（61对）,杨莉莉曹水李慧于津浦任宝柱安秀梅任秀宝天津市肿瘤防治重点实验室天津医科大学附属肿瘤医院研究所免疫室中国肿瘤临床2009年第36卷第17期,28,DC+CIK联合化疗治疗晚期非小细胞肺癌的临床疗效评价,29,Comparativestudyonantitumorimmuneresponseofautologouscytokineinducedkiller(CIK)cells,dendriticcellsCIK(DC-CIK),andsemiallogeneicDC-CIK,QiJingWang,.Jian-chuanXia,StateKeyLaboratoryofOncologyinSouthernChina,Guangzhou,Guangdong510060,P.R.China2DepartmentofExperimentalResearch,SunYatsenUniversityCancerCenter,Guangzhou,Guangdong510060,P.R.ChinaChineseJournalofCancer2010Vol.29Issue7,30,Figure2IntracellularcytokineproductionofCIK,autologousDC-CIK,andsemi-allogeneicDC-CIKcellsA.IFN-gproductionofCIKcells,autologousDC-CIKcells,andsemi-allogeneicDC-CIKcellsfromRCCpatient;B.IL-4productionofCIKcells,autologousDC-CIKcells,andsemi-allogeneicDC-CIKcellsfromRCCpatient.,半相合DC-CIK增加产生IFN-gCIK降低产生IL-4CIK,31,上颌窦鳞状细胞癌,Comparativestudyonantitumorimmuneresponseofautologouscytokineinducedkiller(CIK)cells,dendriticcellsCIK(DC-CIK),andsemiallogeneicDC-CIK,32,NKNatureKiller,33,Naturalkillercell-basedtherapiesFranoisRomagnandEricVivier,FranceF1000MedicineReports2011,3:9,NKcellssenseinteractingcellsviatheiractivatingandinhibitoryreceptors.ThedensityofligandsforthesereceptorsdictateswhetherornotthisinteractionwillleadtoNK-cellactivationandhencecytotoxicityand/orcytokinesecretion.MHC,majorhistocompatibilitycomplex;KIR,killercellimmunoglobulin-likereceptor.,34,Fig1.Protocolschemahaplo-identicalNKcelltherapycombinedwithdelayedautografting.Conditioningtherapycomprisedfludarabine(Flu,25mg/m2onday)5today)2),dexamethasone(Dex,40mg/dondays)5to)2),andmelphalan(MEL,140mg/m2IVonday)1).FluandDexweregiventodepletepatientlymphocytesinordertopreventrejectionofallogeneicNKcells.MEL140mg/m2wasusedfortumourreduction.NKcellsfromahaplo-identicalKIR-ligandmismatchedfamilydonorweretransfusedondays0and+2.IL-2wasgivendailytosupportthesurvivalandexpansionofNKcellsinvivo.AutologousPBSCTwasdelayedtoprovideawindowfordonorNKcellproducttokillresidualmyelomacells,Infusionofhaplo-identicalkillerimmunoglobulin-likereceptorligandmismatchedNKcellsforrelapsedmyelomainthesettingofautologousstemcelltransplantation,KIR错配半相合NK治疗复发骨髓瘤,JumeiShi,etal.,USA,BritishJournalofHaematology,143,641653，2008,氟达拉滨+地塞米松+美法轮,抗CD3磁珠去除T细胞，+IL-2活化扩增NK,35,Infusionofhaplo-identicalkillerimmunoglobulin-likereceptorligandmismatchedNKcellsforrelapsedmyelomainthesettingofautologousstemcelltransplantation,36,CR5+PR1=60%;MR1,SD1,PD1,37,Fig2.AlldonorNKproductskilledKIR-ligandmismatchedMMcells.DonorNKcellslysedMMcelltargetslackinginhibitoryKIR-ligandsincludingpatientMMcells(whenavailable),withtheexceptionofdonor7,whodidnothaveallo-reactiveNKcells.K562,patientMMcells,U266MMcellline(homozygousCgroup1andHLA-Bw4negative),andpatientPHAblastswereemployedastargetsinastandard4-h51Crreleaseassay,atE/Tratiosof10:1.PatientprimaryMMcellswereavailableforpatients2,3,6,7and8.Specificlysispercentagewascalculatedas(testrelease)spontaneousrelease)/(maximalrelease)spontaneousrelease)100.AllexperimentswereperformedintriplicatewellsandthemeanSDwerepresented.Oneofthreeindependentexperimentswasshown.AllexperimentswereperformedwiththefinalNKcellproduct,whichhadbeenincubatedovernight(products14)orduringcellprocessingwith300IU/mlofIL-2(products510).,供者NK杀伤患者骨髓瘤细胞,38,MostofthedonorswerehaploidenticaltothecorrespondingrecipientandatleastoneofthedonorshadaKIRmismatch.DonorsPBbyFicoll-Hypaquedensitycentrifugation,usingstandardprocedures.CD56+cellswereisolatedfromPBMCusingCliniMACSCD56microbeads10-5MHC(氢化可的松),and20ng/mLIL-15,异体NK治疗晚期非小细胞肺癌AphaseItrialofadoptivetransferofallogeneicnaturalkillercellsinpatientswithadvancednon-smallcelllungcancer,EleniG.etal.,Greece,CancerImmunolImmunother(2010)59:17811789,39,异体NK治疗晚期非小细胞肺癌AphaseItrialofadoptivetransferofallogeneicnaturalkillercellsinpatientswithadvancednon-smallcelllungcancer,Fig.1aPercentagesofCD56+CD3-cellsamongtotalCD56+Gatedcellsisolatedfromdonorsperipheralbloodonday0andafter1821daysofculturewith20ng/mLIL-15and10-5MHC(氢化可的松).bExpressionofactivatingNKreceptors(NKp30,NKp44,NKp46,NKG2Dand2B4)ondonorsCD56+cellsexpandedfor1821daysofculturewith20ng/mLIL-15and10-5MHC.Graylinesindicateisotypematchedcontrols.Histogramsfromarepresentativedonor.Alldonorsgavesimilarresults.cCellgrowth(foldincrease)andcytotoxicactivityagainstK562cellsofdonorsCD56+cellsafter2123daysofculturewith20ng/mLIL-15and10-5MHC.Cytotoxicityassaywasperformedataneffector:targetratio1:1.Horizontalbarsindicatemeanvalues,CD56+CD3-cells,40,EleniG.etal.,Greece,CancerImmunolImmunother(2010)59:17811789,2PR12.5%6SD50%8PD,CD56+IL-15,异体NK治疗晚期非小细胞肺癌AphaseItrialofadoptivetransferofallogeneicnaturalkillercellsinpatientswithadvancednon-smallcelllungcancer,41,Fig.2EffectofnumberofinfusionsofallogeneicactivatedNKcellsonprogression-freesurvival(a)andoverallsurvival(b)ofallintendedtotreatnon-small-celllungcancerpatients.cOverallsurvival（OS）ofpatientswithstableordecreasedpulmonarydisease,receiving23and4NKcellinfusions,respectively,异体NK治疗晚期非小细胞肺癌AphaseItrialofadoptivetransferofallogeneicnaturalkillercellsinpatientswithadvancednon-smallcelllungcancer,42,gdT,43,GDT:gdTcells,TTP:timetoprogression,OS:overallsurvival,PD:progressivedisease,SD:stabledisease,BSC:bestsupportivecare,M:moleculartargetedtherapy,C:chemotherapy,R:radiotherapy,I:immunotherapy.aFromthedateof1stinjection.bCase#6,clinicalresponse=SD,after3rdinjection.S,surgery;M,moleculartargetedtherapy;C,chemotherapy;R,radiotherapy;I,immunotherapy.,gdT细胞治疗复发的非小细胞肺癌,AphaseIstudyofadoptiveimmunotherapyforrecurrentnon-small-celllungcancerpatientswithautologousgdTcellsJunNakajima，JapanEuropeanJournalofCardio-thoracicSurgery37(2010)11911197Zoledronate（唑来膦酸）+IL-2.,无放化疗生存长？,SD330%,44,PhaseI/IIstudyofadoptivetransferofTcellsincombinationwithzoledronicacidandIL-2topatientswithadvancedrenalcellcarcinoma,HirohitoKobayashi，Japan，CancerImmunolImmunother，26，April2001,CR1SD554.5%PD5,唑来膦酸+IL-2,45,CTLCytotoxicTLymphocyteTILTumorInfiltratingLymphocyte,46,T细胞过激免疫治疗疫苗免疫-外周血、淋巴结T细胞体外扩增TIL体外扩增,J.Clin.Invest.117(6):1466-1476(2007),外周血淋巴结肿瘤组织,47,HLA-A2+patientswithmetastaticmelanomareceivedimmunodepletingchemotherapywithcyclophosphamideandfludarabine（氟达拉滨）for7daysbeforetheadoptivetransferofhighlyselectedtumor-reactiveTcellsandhigh-doseinterleukin-2(IL-2)therapy,patient9patient10,CancerRegressionandAutoimmunityinPatientsAfterClonalRepopulationwithAntitumorLymphocytes，StevenA.RosenbergSCIENCE2002,298:850,抗肿瘤CTL克隆治疗转移性黑色素瘤,48,PR:6/13(46%)MixedRespose:4/13(30%),Patientdemographics,treatmentsreceived,andclinicaloutcomes,StevenA.RosenbergSCIENCE2002,298:850,49,TIL与放、化疗联合应用治疗转移性黑色素瘤AdoptiveCellTherapyforPatientsWithMetastaticMelanoma:EvaluationofIntensiveMyeloablativeChemoradiationPreparativeRegimensStevenA.RosenbergJClinOncol26:5233-5239.2008,Fig1.(A)Scheduleoftreatmentsadministeredforthreelymphodepletingcelltransferregimens.Cy,cyclophosphamide;Flu,fludarabine;TBI,total-bodyirradiation;CD34,hematopoieticstemcells;TIL,tumor-infiltratinglymphocytes;IL-,interleukin.,化疗和全身放疗是去除抑制细胞,50,TIL与放、化疗联合应用治疗转移性黑色素瘤AdoptiveCellTherapyforPatientsWithMetastaticMelanoma:EvaluationofIntensiveMyeloablativeChemoradiationPreparativeRegimensStevenA.RosenbergJClinOncol26:5233-5239.2008,51,Objectiveclinicalregressionsinpatientswithmetastaticmelanomatreatedwithcelltransfertherapy.(a)Regressionofmelanomametastasesintheheart(upper),adrenal(middle)andperitonealcavity(lower)nowongoingat34monthsina53-year-oldmale.(b)Regressionofmultiplelivermetastasesnowongoingat60monthsina45-year-oldmale.(c)Rapidregressionofmultiplesubcutaneousandnodalmetastasesnowongoingat35monthsina29-year-oldmale.(d)Regressionofalargefungatingscalpmassnowongoingat34monthsina40-year-oldmale.StevenA.Rosenberg，Adoptivecelltherapyforthetreatmentofpatientswithmetastaticmelanoma，CurrentOpinioninImmunology2009,21:233,52,TIL与放、化疗联合应用治疗转移性黑色素瘤,Survivalofpatientstreatedwithcelltransfertherapyinfourconsecutiveclinicaltrialsusingincreasingregimensofalymphodepletingpreparativeregimenbeforeadoptivecelltransfer(NMA,non-myeloablativechemotherapy;TBI,totalbodyirradiation).Thenumberinparenthesesisthenumberofpatientsineachtrial.CurrentOpinioninImmunology2009,21:233240,53,ObjectiveresponseratesusingRECISTcriteriainpatientswithmetastaticmelanomatreatedintheSurgeryBranch,NCIusingdifferenttherapeuticstrategies.Overallresponseratesinpatientstreatedwithvaccinesisabout3%andwithIL-2oranti-CTLA4isabout15%.Withincreasinglevelsoflymphodepletion,addingtotalbodyirradiation(TBI)andnonmyeloablativechemotherapy(NMA)toadoptivecelltransfer(ACT)canachieveresponseratesashighas72%.,TIL与放、化疗联合应用治疗转移性黑色素瘤,实体瘤的疗效评价标准ResponseEvaluationCriteriainSolidTumors,ACT:过继细胞输入NMA:非清髓化疗TBI:全身照射,54,Fig2.Adoptivecelltherapyafterlymphodepletion.(A)Telomerelengthofinfusedtumor-infiltratinglymphocytes(TIL)correlateswithclinicalresponse.Flow-fluorescentinsituhybridizationanalysiswasusedtoquantifythemeantelomerelengthofinfusedTILforpatientsonallthreeprotocols.,AdoptiveCellTherapyforPatientsWithMetastaticMelanoma:EvaluationofIntensiveMyeloablativeChemoradiationPreparativeRegimensMarkE.Dudley,StevenA.Rosenberg,JClinicalOncology,2008,V26(32),5233-5239,过继免疫治疗效果与TIL端粒长短相关,55,改进过继免疫治疗的方法,StevenARosenberg，CurrentOpinioninImmunology2009,21:233240,56,SuicideTcelltherapyusingadoptivelytransferredTcells.Tcellscanbeengineeredtoexpressconditionalsuicideswitchessothatth