免疫學及免疫檢測技術(shù)論文翻譯

Armedandaccurate:engineeringcytotoxicTcellsforeradicationofleukemia裝備并確認：工程細胞毒性T細胞根除白血病MarkoRadicCorrespondence:MarkoRadicmradic@AuthorAffiliationsThisisanOpenAccessarticledistributedunderthetermsoftheCreativeCommonsAttributionLicense(/licenses/by/2.0),whichpermitSunrestricteduse,distribution,andreproductioninanymedium,providedtheoriginalworkisproperlycited.AbstractTranslationalmedicinedependsonarapidandefficientexchangeofresultsbetweenthebenchandthebedside.Arecentexamplefromthefieldofcancerimmunotherapyhighlightstheessentialnatureofthisexchange.Methodshavebeendevelopedtoconvertapatient'scytotoxicTcellsintoefficientandspecifickillersofcancercellsinpatientswithleukemia.ByusingrecombinantDNAtechniques,alentiviralvectorwasconstructedtoexpresschimericantigenreceptorsincytotoxicTcellsfrompatientswithadvancedchroniclymphocyticleukemia.ThepurposeofthechimericreceptorswastodirectthecytotoxicTcellactivityagainstcellscausingthecancer.TheeffectofinfusingtheengineeredTcellsbackintothecancerpatientswastestedinaPhaseItrialattheUniversityofPennsylvania,andtheinitialresultsweredescribedintwoarticlesfromtheresearchteamofDr.CarlJune.Theremarkablesuccessofthistrialshouldenergizefurtherapplicationsofbiotechnologyinthedevelopmentofnewcancerimmunotherapies.轉(zhuǎn)化醫(yī)學取決于工作臺和病床間的快速，高效的轉(zhuǎn)換的結(jié)果。最近的一個例子，從癌癥的免疫療法領(lǐng)域突出這種交換的本質(zhì)。方法已被開發(fā)是將患者的細胞毒性T細胞轉(zhuǎn)換成高效特異的癌細胞殺手。通過使用重組DNA技術(shù)，慢病毒載體構(gòu)建從先進的慢性淋巴細胞白血病的患者的細胞毒性T細胞中表達嵌合抗原受體。嵌合受體的目的是指導細胞毒性T細胞對抗致癌細胞。I期臨床試驗在美國賓夕法尼亞大學設(shè)計的工程T細胞注入到癌癥患者的影響進行了測試，卡爾博士的研究小組的兩篇文章中描述了初步的結(jié)果。這項試驗非凡的成功，應該激勵在新的癌癥免疫療法的發(fā)展中進一步應用生物技術(shù)

MainTextRecentadvancesincancerimmunotherapyhaveallowedtheconversionofTcellsfromleukemiapatientsintoefficientandspecifickillersoftheirowncancercells.Thenewtechniquehasbeenrecentlytestedwithremarkableresults.Theoutwardsignsofthesuccessfultherapyweredramaticandclear.Thecancerpatientsshookwithfever,chillsandpain;theirbloodpressureprecipitouslydropped.TheydriftedinandoutofsleepfordaysfollowingtheinfusionoftheirownengineeredkillerTcells.TheinternalstrugglepeakedbetweentwoandthreeweeksafterTcellreinfusion.Theloadoflysedcancercellsstressedandnearlypoisonedthepatients'kidneys.Theirimmunesystems,reinvigoratedbytheimmunotherapy,haddestroyednearly1kgofcancercells.Nootheravailabletreatmentcouldhaveachievedacomparabletherapeuticsuccess.Thenewtreatment,administeredbyDr.CarlJuneandhiscolleaguesoftheAbrahamsonCancerCenterattheUniversityofPennsylvania,achievedanunprecedentedcompleteeradicationofthecancerintwoofthethreechroniclymphocyticleukemia(CLL)patientsinwhomitwastried.Ithadbeenadreamofmany:theharnessingoftheimmunesystem'sferociouspowertoeliminateanadvancedmetastaticcancer.在腫瘤免疫治療的最新進展已經(jīng)可以轉(zhuǎn)換白血病患者T細胞變成自己的癌細胞的高效特異的細胞殺手。這項新技術(shù)最近已測試成效顯著。成功治療的外在表現(xiàn)是顯著的，明確的。因發(fā)燒，發(fā)冷和疼痛而顫抖的癌癥患者，他們的血壓會急劇下降。在注入他們自己的工程殺傷性T細胞會使他們在睡醒之間輾轉(zhuǎn)反側(cè)好幾天。T細胞回輸后的內(nèi)部斗爭在兩至三個星期間達到高峰。裂解腫瘤細胞的負荷加大了病人的腎臟壓力并幾乎腎中毒。通過免疫治療恢復活力的免疫系統(tǒng)已經(jīng)摧毀了近1公斤的腫瘤細胞。沒有其他可供選擇的治療已經(jīng)取得了類似的治療成功。由卡爾博士和他的在美國賓夕法尼亞大學的亞伯拉罕森癌癥研究中心的同事共同執(zhí)行的新的治療方法，在試驗的3個慢性淋巴細胞白血病(CLL)患者其中兩個實現(xiàn)了前所未有的徹底根除癌癥。它曾是許多人的夢想：利用免疫系統(tǒng)的驚人的治理力量消除晚期轉(zhuǎn)移性癌癥。Thestunningresultswerepublishedintwosimultaneousjournalarticles[1,2],oneofthereportspresentingdatafromasinglepatient[1].Thesuccesswasbeyondexpectation.This,afterall,wasaPhaseIclinicaltrial,oneinwhichtheprimaryobjectivewastotestdifferentdosesoftheengineeredcytotoxiclymphocytes(CTL).Technically,thesuccesscouldbetracedtothefusionofextracellularandintracellulardomainsintoanew,syntheticreceptorfortheCD8+Tcells[3].In_immunotherapyjargon,thistypeofrecombinantproteiniscalledachimericantigenreceptor(CAR).Inthiscase,theextracellulardomainwasderivedfromamousemonoclonalantibodyspecificfortheCD19surfaceproteinofhumanBcells.BecauseCLLisaBcellcancer,CLLcellsexpressCD19.Theanti-CD19antibodywasreducedtoitssmallestactiveform,asingle-chainvariablefragment(scFv),andgraftedontoaTcellreceptortransmembranedomain.Ontheinsideofthecell,theconstructwasfusedtothecytoplasmicsignalingdomainsofCD137(41BB)andtheZhainofCD3(Figure1).Thisparticularcombinationoffunctionaldomainsiscreditedforachievingthespectacularsuccessofthetherapy

令人驚嘆的研究結(jié)果發(fā)表在兩個同時進行的期刊文章，其中一個報告數(shù)據(jù)來自一個單身病人。這個成功的研究是出乎意料的。畢竟，這是第一期臨床試驗，其主要目的之一是測試不同劑量的工程細胞毒性T細胞（CTL）。從技術(shù)上講，這次成功可以追溯到融合細胞外和細胞內(nèi)結(jié)構(gòu)域成一個新的，合成受體的CD8+T細胞。在免疫治療術(shù)語中，這種類型的重組蛋白被稱為嵌合抗原受體（CAR）。在這種案例中，胞外結(jié)構(gòu)域是來自特定的人類B細胞CD19表面蛋白的小鼠單克隆抗體。由于CLL是一種B細胞癌，所以白血病細胞表達CD19蛋白?？?CD19抗體被減少到最小的活性形式，單鏈可變區(qū)片段（scFv）,并且被移植到T細胞受體的跨膜結(jié)構(gòu)域。在細胞的里面，該結(jié)構(gòu)被融合到胞質(zhì)信號域CD137（41BB）和CD3Z鏈（圖1）這種功能結(jié)構(gòu)域的特定組合因獲得治療的驚人成功而得名Truncated—envTruncated-一gag/polWPRE5JLTRTruncated—envTruncated-一gag/polWPRE5JLTRpELPS19-BB-^{11556bp)3,LTR-—BovineVGHPolyA、BacterialReplicationOriginAmpRFigurel.ThebiotechnologyofchimericantigenreceptorsforcytotoxicTcell-basedcancerimmunotherapy.A.MapofpELPS19-BB-3Z，alentiviralvectorthatwasapprovedforclinicaltrials[1].TheopenreadingframeencodingthechimericfusionproteinCD19-BBZcontainsportionscodingforananti-CD19single-chainvariableregionfragment(scFv),thehingeandtransmembrane(TM)domainofCD8,andapairofsignalingendodomains(onefromCD137,theotherfromCD3Z).ThisfusionproteingeneisembeddedwithinadisabledHIV-derivedbackbonecontainingtruncated(CD137Survival1andProliferation(CD137Survival1andProliferationSignalingUrt^1AW%MIMA

gag,polandenvsequencesandthe5'and3'longterminalrepeats（LTR）.Inaddition,theconstructcontainselongationfactorla（EF-1a）codingsequences,thewoodchuckhepatitisviruspost-transcriptionalregulatoryelement（WPRE）andthebovinegrowthhormonepolyadenylation（GHpolyA）tail,aswellastheampicillinresistancegeneandabacterialreplicationorigin.Thetotalsizeoftheconstructis11,556basepairs.B.DiagramofthechimericreceptorproteinintheTcellplasmamembrane.TheextracellularantibodyrecognitiondomainislinkedviatheCD8TMdomaintothecytoplasmicsignalingdomainsofCD137andCD3Z.TheseendodomainsaccomplishtheactivationoftransducedTcellsandtheystimulateTcellsurvivalandproliferationinvivo.圖1.細胞毒性T細胞為基礎(chǔ)的腫瘤免疫治療之嵌合抗原受體的生物技術(shù)。A.pELPS19-BB-3Z圖，慢病毒載體被批準用于臨床試驗的。編碼嵌合融合蛋白CD19-BBZ的開放閱讀框，包含抗CD19單鏈可變區(qū)片段（scFv）,CD8的鉸鏈和跨膜（TM）域，以及一對內(nèi)域信號（一個來自CD137，另一個CD3Z）的部分編碼。此融合蛋白的基因被嵌入一個包含刪除gag,pol和env序列以及5'和3'長末端重復序列（LTR）的帶有缺陷型獲得性HIV脊椎。此外，該結(jié)構(gòu)包含延伸因子1a（EF-1a）的編碼序列，土撥鼠肝炎病毒轉(zhuǎn)錄后調(diào)控元件（WPRE）和牛生長激素多聚腺苷酸化（生長激素多聚A）尾，以及氨芐青霉素抗性基因和一個細菌的復制起點。該結(jié)構(gòu)的總大小是11556個堿基對。B.圖中的T細胞質(zhì)膜的嵌合受體蛋白質(zhì)。通過CD8TM域?qū)崿F(xiàn)胞質(zhì)信號領(lǐng)域的CD137和CD3Z和細胞外抗體識別域的鏈接。這些內(nèi)域完成轉(zhuǎn)導T細胞的激活和刺激T細胞在體內(nèi)的存活和增殖。ThegeneforthisCARwasdeliveredbylentivirustopurifiedcytotoxickillerTcellsfromeachpatient.TheHIV-1virusservedasthebackboneintheconstructionofthelentivirusvector.ThechoiceofthisvirusprovedfortunatebecausethenaturalhostforHIV-1isaTlymphocyte.Therefore,thevectorincludedsequencesthathadevolvedforreplicationandexpressioninTcells.ApreviousversionofthisvectorwasusedtotreatpatientswithchronicHIVinfections[4].TheCLLpatients'purifiedTcellsweretransducedinvitro,thenstoredforthetimerequiredtotreateachpatientwithchemotherapyandreducethenumbersofunmodifiedlymphocytes.ThisprovidedtheinvivospacetoaccommodatetheengineeredTcells.CAR基因通過慢病毒轉(zhuǎn)入純化每個病人的細胞毒性殺傷性T細胞。HIV-1病毒是慢病毒載體的主干。選擇這種病毒是幸運的，因為HIV-1的天然宿主是一種T淋巴細胞。因此，這個載體包含在T細胞中復制和表達的序列。這個載體的早期版本被用于治療艾滋病毒慢性感染。CLL患者的純化的T細胞在體外轉(zhuǎn)導，然后記錄每個化療病人治療和減少未變性淋巴細胞的數(shù)目所需要的時間。這提供了適應工程T細胞在體內(nèi)的空間Oncethepatientswerereadyfortheexperimentaltreatment,theyreceivedbetween15Millionand1BilliontransducedCTL.TheCTLrapidlymultipliedtoapproximately1000-foldhighernumbersinvivo[2]￡artofthisexpansionmaybeattributedtothepresenceoftargetcells.ThescientistsperformingthetrialestimatedthateachengineeredCTLdestroyednearly1000leukemiacells,until,threeweeksaftertheinfusionoftheCTL,nodetectablecancercellsremained.ThepeakofCTLproliferationcorrespondedtothehighestlevelofIFN-Y,IL-6and

CXCL9incirculation[2]Jhesecytokineswereresponsibleforthepatients'highfever.OncetheCLLnumbersdroppedbelowdetectionlevels,theCTLnumbersnormalizedandtheremainingengineeredcellsacquiredthephenotypeofeffectorandmemoryTcells.Atthattime,thepatients'overallwellbeingmarkedlyimproved.一旦患者準備進行實驗性治療，他們收到了15億至10億之間轉(zhuǎn)導的CTL。的CTL迅速乘以在體內(nèi)的約1000倍的更高的數(shù)字[2]。這種擴張的部分內(nèi)容可以歸因于靶細胞的存在。進行試驗的科學家估計，每個工程的CTL破壞近1000個白血病細胞，直到輸注的CTL，三周后，沒有檢測到癌細胞仍然。的CTL增殖的峰值相對應的最高水平的IFN-y，IL-6和CXCL9流通中[2]。這些細胞因子負責病人的高燒。一旦CLL數(shù)字下降到低于檢測水平，CTL數(shù)歸一化和收購余下的工程細胞的表型效應和記憶性T細胞。當時，顯著提高患者的整體健康。Oneside-effectofthetherapyisBcellimmunodeficiencybecausetheengineeredCTLalsokillthepatients'non-cancerousBcells.ThiswasananticipatedconsequenceofthetherapybecausetheCTLtargetantigen,theCD19receptor,isexpressedonallmatureBcellsandBcellprecursors.ThetransferofIgGfromnormaldonors(IVIG)canreplenishthepatients'circulatingantibodiesandpartiallycompensatefortheabsenceofBcells.ThelgGinjectionswillbeperformedatmonthlyintervalstoreducetheriskofinfectionsintreatedpatients.Twoofthepatientswere,atthetimeofthereports,freeofcancerrecurrencefor6to11months[2].ThethirdshowedasustainedimprovementoftheCLL.治療的一個副作用是B細胞免疫缺陷，因為工程CTL也殺死患者的非癌性的B細胞。這是一個預期的治療結(jié)果，因為CTL的靶抗原，CD19受體被表達于所有成熟B細胞和B細胞的前體。來自正常捐助者(IVIG)的IgG轉(zhuǎn)移可以補充抗體循環(huán)并局部彌補缺乏B細胞。將進行間隔每月一次的IgG注射來減少治療的患者中感染風險。當時的報道，患者中有兩位在6個月至11個月無癌癥復發(fā)。第三個表現(xiàn)出CLL持續(xù)改善VariousgroupsofscientistshadattemptedtomodifyCTLtoattackcancercells.Inpioneeringwork,EshharandcollaboratorsfromtheWeizmannInstituteofScienceinRehovot(Israel)demonstratedthatCTLcanexpressarecombinantsurfacereceptorthatlinksanextracellularrecognitiondomainderivedfromanantibodytoanintracellularsignalingdomainderivedfromtheTcellreceptor[5].SuchTcellsnolongerdependonrecognitionofHLA-peptidecomplexesforspecificsignalingandTcellactivation.各種科學家團體曾試圖修改CTL來攻擊癌細胞。在開創(chuàng)性的工作中，Eshhar和來自以色列魏茨曼科學研究所的合作者表明，CTL表達的重組表面受體連接獲得抗體的胞外識別域和獲得T細胞受體的細胞內(nèi)信號域。這種T細胞不再依賴于特定的信號和T細胞活化的HLA-肽復合物識別。Normally,cytotoxicTcellsrecognizeforeignpeptidesinthegrooveofanHLAclassIreceptor.Thepeptidesmaybederivedfromavirusinfectingahostcell,whichdegradesaportionofthenewlyformingviralproteinsintopeptidesandloadsthemintoaclassIHLA.TherecognitionofthepeptidesasforeigntothehosttriggerstheTcelltoreleaseperforinandgranzyme,proteinsthatformporesinthetargetcellandinitiateacascadeofenzymaticreactionsleadingtothe

programmeddeathoftheinfectedcells.Inthismanner,thevirusispurgedfromthebody.通常情況下，細胞毒性T細胞在HLA類的受體的凹槽中識別外源肽。從病毒感染的宿主細胞中可以獲得這些降低部分新形成的病毒蛋白形成的多肽并加載它們?yōu)镮類HAL。從外來得到宿主的多肽的識別觸發(fā)T細胞釋放穿孔素和顆粒酶，蛋白質(zhì)，形成在目標小區(qū)中的孔，并啟動酶促反應，導致受感染的細胞程序性死亡的級聯(lián)。在這種方式中，該病毒是從體內(nèi)清除AnanalogousreactionisthoughttooccurbetweencytotoxicTcellsandtumorcells.Theoncogenictransformationofacancercellinducestheexpressionofproteinsthatarenotexpressedinhealthytissues.TheimmunesystemcandetectthedifferencesinHLA-associatedpeptidesfromcancercells,andthecytotoxicTcells,inconsequence,releasetheircell-degradingeffectormolecules.However,inadvancedcancers,thesecytotoxicTcellsaredepletedandinefficient.Gradually,thecancercellsgaintheupperhandandgrowtonumbersthatbecomemoreandmoredifficulttocontrol.一個類似的反應被認為是發(fā)生在細胞毒性T細胞和腫瘤細胞之間。癌癥細胞的致癌性轉(zhuǎn)化誘導在健康組織中不表達的蛋白質(zhì)的表達。免疫系統(tǒng)可以檢測癌細胞中與HAL相關(guān)的多肽的差異，因此，細胞毒性T細胞釋放其降解細胞的效應分子。然而，在晚期癌癥，這些細胞毒性T細胞被耗盡，效率不高。漸漸地，癌細胞占據(jù)上風和成長的數(shù)字變得越來越難以控制InTcellsexpressingchimericantigenreceptors,thescFvantibodyfragmentthatbindstoantigensubstitutesfortheTCRbindingtoHLApluspeptide.TheinitialsuccessofGrossetal.[5]inspired_numeroussubsequentstudiesbutobstacleshinderedeffortstoexpressrecombinantreceptorsthatcouldbindthedesiredtargetsandactivatethecytolyticfunctionsoftheTcells[6].VariousinvestigatorsdesignedandtestedsecondandthirdgenerationCARfusionproteinsbecausesingleactivationdomainCARshowedonlylimitedutilityforthestimulationofnaiveTcells[6]^Themoreadvancedreceptordesignincorporatedadditionalendodomainsthatcouldsimultaneouslyactivatemorethanonesignalingpathway.ExperimentsbyImaietal.[7]and_Finneyetal.[8]testeddualactivationdomains,includingthecombinationofCD3andCD137(4-1BB)andfoundtheaddedactivationdomainstoincreaseresponsesofengineeredCTL.Subsequently,Miloneetal.[3]demonstratedtheinvivoefficacyofthenewchimericactivationdomainsinxenogeneictumorbearingmice.TheseexperimentssetthestageforthesuccessfulclinicaltrialattheUniversityofPennsylvania[1,2].在T細胞中表達嵌合抗原受體，scFv抗體片段綁定的結(jié)合HLA加肽的TCR抗原代用品。Gross等人的初步成功鼓勵了眾多的后續(xù)研究，但可以結(jié)合預期的目標和激活的T細胞的殺傷功能的重組受體的表達受到阻礙。各種研究人員設(shè)計和測試了第二代和第三代的CAR融合蛋白，因為單激活域CAR僅表現(xiàn)對幼稚T細胞的刺激的有限用途。更先進的受體設(shè)計還采用了其它內(nèi)域能同時激活多個信號轉(zhuǎn)導通路。Imai等人和芬尼等人的實驗測試了雙激活域，包括CD3和CD137(4-1BB)相結(jié)合的測試，發(fā)現(xiàn)增加對工程CTL效應的其他激活域。接著，Milone等表明在異種荷瘤小鼠的新的嵌合激活域在體內(nèi)的療效。在賓夕法尼亞大學，這些實驗成功的臨床試驗階段。

ThebreakthroughoccurredwiththediscoverythatthecytoplasmicdomainfromCD137(4-1BB)enabledstrongactivationoftheengineeredTcells,providedthatitwasinsertedbetweentheCD8atransmembraneregionandtheCD3Zcytoplasmicdomain[3].ThistandemconfigurationofactivatingdomainsmergedintoasinglepolypeptidechainthetaskofdeliveringtwosignalsneededfortheactivationofnaiveTcells(Figure1).Inthismanner,bindingoftheextracellularantibodydomaintoantigencouldtransmittwosignalsneededtoinducethematurationofnaiveCD8TcellsintoeffectorTcells.Thecellsexpressingthetandemchimericantigenreceptorsreceivedstrongsignalstoproliferateanddifferentiateintopotentcancer-destroyingkillerTcells[1].這一突破來自CD137(4-1BB)的胞質(zhì)域中使工程T細胞增強活性的發(fā)現(xiàn)，條件是它被插入CD8a跨膜區(qū)和CD3Z胞質(zhì)域之間的。這種串聯(lián)結(jié)構(gòu)的激活提供幼稚T細胞的活化(圖1)所需的兩個信號的任務合并成一個單一的多肽鏈的域。以這種方式，對抗原的抗體的胞外域綁定可以傳輸需要兩個信號誘導幼稚CD8+T細胞的成熟成效應T細胞。串聯(lián)嵌合抗原受體的細胞表達獲得強烈的信號進行增殖并分化成強力摧毀癌癥的殺傷性T細胞。Clearly,thenewresearchwillstimulateeffortstoextendthisapproachtootherrefractorycancers.Still,caveatsremain.Thenumbersofpatientstreateduptonowremainverysmallandthetimeelapsedsincethetreatmentisshort,thussuccesscouldstillremainelusive.However,hopesthatapromisingnewtherapyiswithinreachincreasewitheverypassingday.Inall,morethan25clinicaltrialscurrentlyexploretheuseofchimericTcellreceptorsinvariousformsofcancer[9].Itmight_befeasibletoextendtheCLLtherapy[1,2]tootherBlymphocyticneoplasms,assimilarlyengineeredCTLshouldbecapableofattackinganyCD19-positivecancercells.Yetimportanthurdlesremain:Itmayprovedifficulttodevelopantibodieswithexclusivespecificityforcancercellsthatdonotcross-reactwithhealthytissues.Anycross-reactivitymayleadtoautoimmunitythatcoulddamagevitalorgans.Furthermore,theviolentside-effectsassociatedwiththeclearanceofcancercellsmakeitdesirabletofindwaystoregulatetherateofCTLactivation.Forexample,therearewaysinwhicha"safetyswitch"maybeincorporatedintoengineeredTcellsthatcoulddeactivatethemincasethattheCTLtherapyexposespatientstounacceptablecomplications[10].ThesuccessofthephaseItrialwithtandemendodomainCARsdeliveredbylentivirus[1.2]willsurelyenergizethesearchforadditionalrecombinantimmunetherapiesforadvancedcancer.顯然，新的研究將刺激拓展這種方法到其他難治性癌癥的努力。不過，注意事項依然存在。到現(xiàn)在治好的患者人數(shù)仍然非常少并且療程短，因此成功可能仍然是難以捉摸的。然而，有前途的新療法的希望是日趨增加。超過25項臨床試驗，目前正在探索各種形式的癌癥。它可能是可行的，延長CLL的治療到其他B淋巴細胞腫瘤，像類似的工程CTL應該是能夠攻擊任何CD19陽性癌細胞的。然而，重要的障礙依然存在：它可能難以對不與健康組織發(fā)生交叉反應的腫瘤細胞產(chǎn)生專一特異性抗體。任何交叉反應可能會導致自身免疫性疾病，可能會損壞重要器官。此外，與癌細胞的清除的嚴重副作用使得必須能找到方法來調(diào)節(jié)CTL的激活率。例如，有一些方法，其中一個能禁止“安全開關(guān)”成為工程T細胞一部分以防CTL治療使患者遭受不可接受的并發(fā)癥。通過慢病毒傳遞的帶有串聯(lián)內(nèi)域CARs的I期臨床試驗的成功無疑必將激勵尋找對晚期癌癥其他的重組免疫療

CompetinginterestsTheauthordeclaresthathehasnocompetinginterests.AcknowledgementsTheauthoracknowledgestheexpertassistanceofDafneSolera,Ph.D.,ExecutiveEditorofBMCBiotechnology,withessentialadjustmentsinstyleandcontent.ThetechnicaldiagramofMr.TimHiggins,seniorillustrator,isacknowledged.ReferencesPorterD丄evineBL,KalosM,BaggA,JuneCH:Chimericantigenreceptor-modifiedTcellsinchroniclymphoidleukemia.NEnglJMed2011,365:725-733.PubMedAbstract|PublisherFullTextKalosM,LevineBL,PorterDL,KatzS,GruppSA,BaggA,JuneCH:Tcellswithchimericantigenreceptorshavepotentantitumoreffectsandcanestablishmemoryinpatientswithadvancedleukemia.SciTranslMed2011,3:95ra73.PubMedAbstract|PublisherFullTextMiloneMC,FishJD,CarpenitoC,CarrollRG,BinderGK,TeacheyD,SamantaM,LakhalM,GlossB,Danet-Des