白介素12的创新临床应用肿瘤免疫治疗的新思路

ANewApproachtoCancerImmunotherapy肿瘤免疫治疗的新思路斯坦福大学医学院外科宗康拉1)Slowertumorgrowth(抑制90%)2)Tumorsshrink(肿瘤体积减小,少见)3)Tumorsdisappear(治愈,极少)治疗0-30days5x105tumorcellsHowtodoantitumorexperiments如何做肿瘤治疗效果试验32-dayMCA207beforetreatmentOneweekafterCy+IL-12TwoweeksafterCy+IL-12ThreeweeksafterCy+IL-12FourweeksafterCy+IL-12FiveweeksafterCy+IL-12SixweeksafterCy+IL-12PotentiationofCy-inducedCancerRegressionbyIL-12通过白介素12提高化疗的抗肿瘤效果noRxCyaloneIL-12aloneCy+IL-12Day-18peritonealMCA207tumors18天腹腔实体MCA207肿瘤Day-14experimentallungmetastases14天静脉注射建立的肺扩散肿瘤模型AntitumoreffectsofCy+IL-12inMCA207i.p.andi.v.modelsCy+IL-12在腹腔及肺扩散肿瘤模型中的治疗效果routeofinoculation接种treatment(startatday)治疗（起始时间）curerate(survivaldays)治愈率（存活天）i.p.腹腔i.p.腹腔i.v.肺扩散None无Cy+IL-12(18)Cy+IL-12(14)0/5(20-27)0/5(21-31)5/5(90)8/8(90)i.v.肺扩散None无EffectsofIL-12andCy+IL-12intheSa1ascitestumormodelIL-12和Cy+IL-12在Sa1腹水肿瘤模型中的治疗效果100%50%0%saline生理盐水IL-12CyCy+IL-12010203040tumor接种treatment治疗survival存活率days天数TherejectioninducedbyIL-12/Cy+IL-12isassociatedwithastrongTcellresponse与肿瘤排斥所对应的强免疫反应CD4CD8before之前after之后TumorrejectionismediatedbyaTh1response抗肿瘤作用需要Th1型T细胞参与host宿主curerate治愈率Normal正常TCRbKOT细胞受体敲除Nude裸鼠IFN-gKO咖玛干扰素敲除IL-4KO白介素-4敲除10/100/30/200/2010/10DoesCy+IL-12workonothertumormodels?环磷酰胺加白介素是否对所有肿瘤有效？Respondingtumors:C57B/6:MC203,MCA205,MCA207,FBL-3BALB/c:CT26,CSA1M,OV-HMA/J:Sa1Non-respondingtumors:C57BL/6:MCA101,B16,LLC,Pan02,EL-4BALB/c:4T1,S180Question问题Ifimmunotherapyisabletoeradicatelate-stagelargetumorburdens,whatistheproperconditionforit?如果免疫疗法有可能治愈晚期癌症，条件是什么？DoesCy+IL-12workonothertumormodels?环磷酰胺加白介素12是否对所有肿瘤有效？tumor/straincureratetumorsize(day)MCA203/B6MCA105/B6MCA205/B6Sa1/AB6F1CT-26/CB6F1MCA101/B6B16/B6Lewislung/B6Pan02/B6immunogenicyesyesyesyesyesnonononono13-17mm(14)10-14mm(14)9-15mm(14)12-22mm(10-14)10-13mm(14)7-15mm(10-14)4-13mm(7-14)4T1/CB6F15-14mm(7-14)6-10mm(14)5-17mm(7-12)5/54/54/512/156/100/100/100/100/100/5Condition#1：Pre-existingimmunity条件一：预存免疫Whatispre-existingimmunity?什么是预存免疫？•Antigen－specificrecognitionoftumorbythehostimmunesystem宿主对肿瘤抗原有特异性识别•Theimmunesystemhasrespondedtotheexistingtumorpriortotherapystart宿主免疫系统在治疗之前已经对肿瘤有攻击•Thehostresponsetothetumoriscell-mediatedTh1type宿主对肿瘤的应答属于Th1型细胞反应Experimentalprocedureforadoptivecelltransfer体细胞转导试验步骤donor供体tumorvaccine瘤苗tumorchallenge肿瘤接种tumor-free排斥接种recipient受体tumorchallenge肿瘤接种14dayTcelltransfer输入T细胞TcellIL-12/Cy+IL-12治疗Response效果?2dayTumor-sensitizedTcellsarenecessaryforIL-12-inducedtumorrejection肿瘤特异的T细胞在白介素12治疗中的关键作用donorcells输入细胞treatment治疗curerate治愈率nonenaïveTcellstumor-immuneTcellsIL-12salinesalineIL-12IL-12Cy+IL-120/10assemblyofpre-existingimmunityinTcell-deficienthost0/80/81/109/1210/10tumor-immuneTcellsnaïveTcellstumor-immuneTcellsnaïveTcellsCy+IL-120/5Ccondition#2条件2IL-12shouldbegivenduringtheearlyphaseofrecalledpre-existingimmunity白介素12最佳给药时间是在预存免疫的回放早期TimingofIL-12followingchemotherapy白介素12的给药最佳时期timetumorsizeCyIL-12IL-12IL-12CriticaltimingofIL-12administration白介素12给药时间的关键性IL-12timingfollowingCycurerate治愈率day3-7（第3－7天）100%day7-11第7－11天）day14-18（第14－18天）40%0%LargeMCA207modelCyat125mg/kgIL-12at200ngx3(q.o.d.)Ccondition#3条件3chemotherapymustactivateantitumorimmunity化疗必须激活一个抗肿瘤免疫反应Chemotherapytoactivateantitumorimmunity?化疗引发抗肿瘤免疫反应？ImmunityisresponsibleforcureofsmalltumorbyCychemotherapy免疫参与是环磷酰胺化疗治愈小肿瘤的必要条件host70-100%tumorburdencureratenormalnormalnoTcell3-day7-10-day8-day0%0%MCA207smalltumormodelused3-daytumorisnon-palpable7-10tumorsare2-5mminsizeCyat125mg/kgisusedPEIstatusnotyetestablishedneverImmunityisresponsibleforsignificantlargetumorregressionfollowingchemotherapy免疫参与加大环磷酰胺化疗疗效CytimetumorsizenormalnoTcellWhycanchemotherapyactivateantitumorimmunity?为什么化疗可以激活抗肿瘤免疫反应？Throughacuteantigenreleaseandrecallofpre-existingimmunity通过抗原急性释放达到记忆免疫的回放Iftrue,thenincreaseantigenpresentationatthesiteofantigenreleasemayincreaseresponsetochemotherapy如果如此，那么在抗原释放位置增加抗原呈递救有可能提高化疗疗效IncreaseantigenpresentationbyDCfollowingchemotherapyenhancestumorresponses提高化疗后的抗原呈递可以提高化疗疗效curerate治愈率treatment治疗Cyalone单独化疗DCalone单独树突细胞Cy+DC联合治疗Responses应答regr.relapse2/10progressionregression0/1010/10Mediumsized(8-11mm)MCA207tumorused.Cy=120mg/kggivenonday21DC=culturedimmatureDCat1x106givenintratumortwodaysafterCyChemotherapyResponses,RelapseandResistancetoRepeatedTherapy化疗应答，复发及随后的抗药性RelapsedtumorisresistanttorepeatedchemotherapywithCyinnormalmice肿瘤复发后对二次化疗产生抗药性1stCytimetumorsize2ndCynormalnoTcellRelapsedtumorfollowingCybecomesresistanttoCy+IL-12therapy复发后的肿瘤对Cy＋IL-12也产生抗药性micebearingresponsetoCy+IL-12CyaloneuntreatedtumorCy-treatedrelapsedtumor+++100%cure+20%cureMCA207largetumormodelused+++:significanttumorregressionfollowingCy+:transientshallowtumorregressionfollowingCyresponsetoCy+IL-12:curebystandardCy+IL-12therapyWhatisthereasonforrelapse-associatedresistancetorepeatedchemotherapy?产生抗药性的原因是什么呢？MDR?Selectionofchemo-resistanttumorcellisNOTthereasonforresistance抗药性不是由于筛选了抗药性肿瘤细胞的原因tumorfromsensitivitytoCyafterreplantinginnaïvehostuntreatedstockafter1stCyrelapseafter2ndCyrelapse100%100%100%SensitivitytoCyinvivoismeasuredbycomplet