腫瘤的免疫細胞療法摘要：隨著人們壽命的增加，癌癥逐漸成為了人類死亡的主要原因之一。針對癌癥的治療也一直是全球的研究熱點。傳統(tǒng)治療腫瘤的手術(shù)療法，放療，化療并不能從根本上消滅腫瘤細胞，往往有很高的復發(fā)率。而腫瘤的免疫療法為治愈癌癥帶來了新的希望。本文對免疫療法發(fā)展過程中出現(xiàn)的各種細胞療法進行了介紹。關(guān)鍵詞：免疫療法LAKTILCIKDC-CIKCAR-T根據(jù)世界衛(wèi)生組織2014年世界癌癥報告，癌癥是全球主要原因死亡原因之一，2012年約新增癌癥病例1400萬，癌癥相關(guān)死亡病例820萬。面對嚴峻的形勢，全球投入了大量的人力和資源研究腫瘤產(chǎn)生機制以及預防和治療的方法。一直以來，腫瘤的臨床治療手段主要是手術(shù)、放療和化療，雖然有一定的效果，但很難從根本上達到治療和控制腫瘤轉(zhuǎn)移和復發(fā)的目的。近年來，腫瘤的免疫療法的發(fā)展為解決這一難題帶來了希望。免疫療法的根源可以上溯到1897年，化學治療的建立者PaulEhrlich提出“魔術(shù)子彈”概念（magicbulletconcept）StrebhardtK,UllrichA.PaulEhrlich’smagicbulletconcept:100yearsofprogress.NatRevCancer,2008,8(6):473–480.。他認為，靶向藥物理論上能夠有效地攻擊病原體而對健康的組織無害，并且提出免疫系統(tǒng)能夠防止腫瘤發(fā)生，抗體是一種能確定目標而不傷害機體的“魔術(shù)子彈”。然而他并沒在腫瘤細胞上找到特定的免疫原性受體。因此癌癥的研究也沒有成為他的主要工作。Ehrlich提出的靶向治療理念并不是嚴格上的免疫療法，因為很多化學藥物對于生物分子或細胞也有靶向的作用，直到1875年“雜交瘤”制備單克隆抗體技術(shù)的發(fā)明K?hlerG,MilsteinC.Continuousculturesoffusedcellssecretingantibodyofpredefinedspecificity.Nature,1975,256(5517):495–497.才奠定了免疫療法的基礎。免疫療法的另一基石是Burnet和Thomas提出“免疫監(jiān)視”（immunesurveillance）概念。他們認為免疫系統(tǒng)一直在監(jiān)視著體內(nèi)的各種細胞，一旦出現(xiàn)癌變就會啟動殺傷功能，而腫瘤的發(fā)生是免疫系統(tǒng)失調(diào)造成的BurnetM.Cancer:Abiologicalapproach.I.Theprocessesofcontrol.BrMedJ,1957,1:779–786。然而裸鼠模型的研究發(fā)現(xiàn)，免疫系統(tǒng)缺陷的小鼠腫瘤發(fā)生率并未顯著提高StutmanO.Tumordevelopmentafter3-methylcholanthreneinimmunologicallydeficientathymicnudemice.Science,1974,183:534–536。這一發(fā)現(xiàn)打擊了“免疫監(jiān)視”概念，也使得免疫療法的發(fā)展出現(xiàn)了停滯。但隨著眾多腫瘤特異性抗原的發(fā)現(xiàn)和T細胞受體（Tcellreceptor,TCR）McIntyreBW,AllisonJP.ThemouseTcellreceptor:structuralheterogeneityofmoleculesofnormalTcellsdefinedbyxenoantiserum.Cell,1983,34:739–746、共刺激因子JuneCH,LedbetterJA,GillespieMM,LindstenT,ThompsonCB.T-cellproliferationinvolvingtheCD28pathwayisassociatedwithcyclosporine-resistantinterleukin2geneexpression.MolCellBiol,1987,7:4472–4481.、抗原提呈細胞（antigen-presenting

cell,APC）JenkinsMK,SchwartzRH.Antigenpresentationbychemicallymodifiedsplenocytesinducesantigen-specificTcellunresponsivenessinvitroandinvivo.JExpMed,1987,165:302–319等免疫作用機制的發(fā)現(xiàn)是免疫治療的研究再次得到發(fā)展。臨床上第一次利用單克隆抗體治療腫瘤是1980StrebhardtK,UllrichA.PaulEhrlich’smagicbulletconcept:100yearsofprogress.NatRevCancer,2008,8(6):473–480.K?hlerG,MilsteinC.Continuousculturesoffusedcellssecretingantibodyofpredefinedspecificity.Nature,1975,256(5517):495–497.BurnetM.Cancer:Abiologicalapproach.I.Theprocessesofcontrol.BrMedJ,1957,1:779–786StutmanO.Tumordevelopmentafter3-methylcholanthreneinimmunologicallydeficientathymicnudemice.Science,1974,183:534–536McIntyreBW,AllisonJP.ThemouseTcellreceptor:structuralheterogeneityofmoleculesofnormalTcellsdefinedbyxenoantiserum.Cell,1983,34:739–746JuneCH,LedbetterJA,GillespieMM,LindstenT,ThompsonCB.T-cellproliferationinvolvingtheCD28pathwayisassociatedwithcyclosporine-resistantinterleukin2geneexpression.MolCellBiol,1987,7:4472–4481.JenkinsMK,SchwartzRH.Antigenpresentationbychemicallymodifiedsplenocytesinducesantigen-specificTcellunresponsivenessinvitroandinvivo.JExpMed,1987,165:302–319NadlerLM,StashenkoP,HardyR,etal.Serotherapyofapatientwithamonoclonalantibodydirectedagainstahumanlymphoma-associatedantigen.CancerRes,1980,40(9):3147–3154.MaloneyGD,GrilloLópezAJ,WhiteCA,etal.IDEC-C2B8(Rituximab)anti-CD20monoclonalantibodytherapyinpatientswithrelapsedlow-gradenon-Hodgkin’slymphoma.Blood,1997,90(6):2188–2195.陳雪靜.抗腫瘤抗體藥物的研究進展.中國生物制品學雜志,2015,28(1):84-90.除了直接注射抗體、細胞因子等激活人體自身免疫系統(tǒng)外，還有一類免疫療法通過直接向病人輸入激活的免疫細胞來治療癌癥，又稱為“免疫細胞療法”或“細胞療法”。免疫細胞療法到目前為止至少有以下五種類型：淋巴因子激活的殺傷細胞（lymphokineactivatedkillercells,LAKcells）療法LAK細胞首次發(fā)現(xiàn)是通過腫瘤患者外周淋巴細胞在含有高劑量的白介素-2（interleukin-2，IL-2）的培養(yǎng)液中短期培養(yǎng)誘導產(chǎn)生，能夠殺傷新鮮分離的實體瘤細胞和體外培養(yǎng)的瘤細胞GrimmEA,MazumderA,RosenbergSA.ThehumanIl-2inducedkillercellphenomenon-lysisofNkresistantfreshsolidtumor-cellsbyIl-2-activatedPbl.FedProc,1982,41:957。LAK是一種T細胞和自然殺傷細胞（naturalkillercells，NKcells）組成的混合物。在體外實驗中，這兩種細胞都表現(xiàn)出了對于廣譜腫瘤細胞的非主要組織相容性復合體限制性（non-majorhistocompatibilitycomplex-restricted，non-MHC-restricted）的細胞毒性PhillipsJH,LanierLL.Dissectionofthelymphokine-activatedkillerphenomenon.RelativecontributionofperipheralbloodnaturalkillercellsandTlymphocytestocytolysis.JExpMed,1986,164:814–825.。將癌癥病人的外周淋巴細胞體外誘導擴增成LAK細胞回輸后，腫瘤出現(xiàn)消退RosenbergSA,LotzeMT,GrimmEA,MazumderA,RosenbergSA.ThehumanIl-2inducedkillercellphenomenon-lysisofNkresistantfreshsolidtumor-cellsbyIl-2-activatedPbl.FedProc,1982,41:957PhillipsJH,LanierLL.Dissectionofthelymphokine-activatedkillerphenomenon.RelativecontributionofperipheralbloodnaturalkillercellsandTlymphocytestocytolysis.JExpMed,1986,164:814–825.RosenbergSA,LotzeMT,MuulLM,etal.Observationsonthesystemicadministrationofautologouslymphokine-activatedkillercellsandrecombinantinterleukin-2topatientswithmetastaticcancer.NEnglJMed,1985,313:1485–1492.陳國友.LAK細胞識別、結(jié)合及殺傷腫瘤細胞分子機制的研究進展.國外醫(yī)學免疫學分冊,1994,3:123-127.AtkinsMB,GouldJA,AllegrettaM,etal.PhaseIevaluationofrecombinantinterleukin-2inpatientswithadvancedmalignantdisease.JClinOncol,1986,4:1380–1391.ThatcherN,DazziH,JohnsonRJ,etal.Recombinantinterleukin-2(rIL-2)givenintrasplenicallyandintravenouslyforadvancedmalignantmelanoma.AphaseIandIIstudy.BrJCancer,1989,60:770–774.腫瘤浸潤淋巴細胞（tumor-infiltratinglymphocytes，TIL）療法1863年，RudolfVirchow首次在腫瘤組織中發(fā)現(xiàn)淋巴細胞Virchow,R.,1863.CellularPathology.Philadelphia.。Zettergren第一次從腫瘤組織中分離出淋巴細胞ZettergrenJG,LuberoffDE,Pretlow2ndTG.Separationoflymphocytesfromdisaggregatedmousemalignantneoplasmsbysedimentationingradientsofficollintissueculturemedium.J.Immunol,1973,111:836-840.。從多種小鼠腫瘤中分離培養(yǎng)的淋巴細胞被證明有體內(nèi)抗腫瘤活性RosenbergSA,SpiessP,LafreniereR.Anewapproachtotheadoptiveimmunotherapyofcancerwithtumorinfiltratinglymphocytes.Science,1986,233:1318-1321.。TIL細胞具有良好的腫瘤特異性，主要作用于CD3+CD56-CD8+的細胞，只能殺死自體同源的腫瘤細胞而對于不同Virchow,R.,1863.CellularPathology.Philadelphia.ZettergrenJG,LuberoffDE,Pretlow2ndTG.Separationoflymphocytesfromdisaggregatedmousemalignantneoplasmsbysedimentationingradientsofficollintissueculturemedium.J.Immunol,1973,111:836-840.RosenbergSA,SpiessP,LafreniereR.Anewapproachtotheadoptiveimmunotherapyofcancerwithtumorinfiltratinglymphocytes.Science,1986,233:1318-1321.RosenbergSA,YannelliJR,YangJC,TopalianSL,SchwartzentruberDJ,WeberJS,etal.Treatmentofpatientswithmetastaticmelanomawithautologoustumor-infiltratinglymphocytesandinterleukin2.JNatlCancerInst,1994,86:1159.RosenbergSA,YangJC,SherryRM,KammulaUS,HughesMS,PhanGQ,etal.DurablecompleteresponsesinheavilypretreatedpatientswithmetastaticmelanomausingT-celltransferimmunotherapy.ClinCancerRes,2011,17:4550.過去十年間，TIL療法的作用機制得到了很好的闡明，尤其是宿主淋巴缺失的條件GattinoniL,FinkelsteinSE,KlebanoffCA,etal.Removalofhomeostaticcytokinesinksbylymphodepletionenhancestheefficacyofadoptivelytransferredtumor-specificCD8+Tcells.JExpGattinoniL,FinkelsteinSE,KlebanoffCA,etal.Removalofhomeostaticcytokinesinksbylymphodepletionenhancestheefficacyofadoptivelytransferredtumor-specificCD8+Tcells.JExpMed,2005,202,907-912.YaoX,AhmadzadehM,LuYC,etal.LevelsofperipheralCD4(+)FoxP3(+)regulatoryTcellsarenegativelyassociatedwithclinicalresponsetoadoptiveimmunotherapyofhumancancer.Blood,2012,119:5688-5696.ChaconJA,SarnaikAA,ChenJQ,etal.2015.Manipulatingthetumormicroenvironmentexvivoforenhancedexpansionoftumor-infiltratinglymphocytesforadoptivecelltherapy.ClinCancerRes,2005,21:611-621.細胞因子誘導的殺傷細胞（cytokine-inducedkillercells，CIKcells）療法CIK細胞是是一類體外誘導產(chǎn)生的異質(zhì)細胞，兼具T細胞的抗腫瘤活性和NK細胞非特異性殺瘤特點，通過刺激和恢復免疫系統(tǒng)對腫瘤細胞識別和殺傷MaY,XuYC,TangL,ZhangZ,WangJ,WangHX.Cytokine-inducedkiller(CIK)celltherapyforpatientswithhepatocellularcarcinoma:efficacyandsafety.ExpHematolOncol,2012,1:11。與LAK細胞類似，CIK細胞也是通過體外培養(yǎng)外周血淋巴細胞產(chǎn)生，除了添加IL-2外，CIK細胞的誘導還添加了一些其他因子，如IL-1，IFN-γ，Abanti-CD3Schmidt-WolfIG,NegrinRS,KiemHP,etal.UseofaSCIDmouse/humanlymphomamodeltoevaluatecytokine-inducedkillercellswithpotentantitumorcellactivity[J].JExpMed,1991,174(1):139-149.。相對于LAK細胞和MaY,XuYC,TangL,ZhangZ,WangJ,WangHX.Cytokine-inducedkiller(CIK)celltherapyforpatientswithhepatocellularcarcinoma:efficacyandsafety.ExpHematolOncol,2012,1:11Schmidt-WolfIG,NegrinRS,KiemHP,etal.UseofaSCIDmouse/humanlymphomamodeltoevaluatecytokine-inducedkillercellswithpotentantitumorcellactivity[J].JExpMed,1991,174(1):139-149.1、直接殺傷作用。CIK對靶細胞的殺傷可能有兩種途徑。一是CIK在淋巴細胞功能相關(guān)抗原-1（lymphocytefunctionassociatedantigen-1,LFA-1）的識別結(jié)常組織受損DuongCP,WestwoodJA,BerryLJ,etal.EnhancingthespecificityofT-cellsculturesforadoptiveimmunotherapyofcancer