腫瘤免疫研課程班課件

腫瘤免疫TumorImmunity腫瘤抗原機(jī)體抗腫瘤的免疫機(jī)制腫瘤的免疫逃逸機(jī)制腫瘤的免疫治療

Modelofsequentialgeneticalterationsleadingtometastaticcoloncancer.Eachofthestagesindicatedatthebottomismorphologicallydistinct,allowingresearcherstodeterminethesequenceofgeneticalterations.屢次打擊模型(MultihitModel)一、腫瘤特異性抗原二、腫瘤相關(guān)性抗原腫瘤抗原

(TumorAntigens)化學(xué)和物理致癌因素誘發(fā)的腫瘤抗原病毒誘發(fā)的腫瘤抗原癌基因和突變型抑癌基因表達(dá)的腫瘤抗原正常靜止基因表達(dá)的腫瘤抗原一、腫瘤特異性抗原

Micewereinducedtoproducetumorsbytheinjectionofachemicalcarcinogen(methyl-cholanthrene).Tumorcellsfromthesemiceweretheninjectedsubcutaneouslyintogeneticallyidenticalmice.Later,thegrowingtumorwereremovedsurgically.Micechallengedwiththesametumorwereabletorejectit,butthosechallengedwithadifferenttumor(inducedwiththesamecarcinogen)werenot.Theabilitytorejectthetumorcouldbetransferredwithlymphoidcells.化學(xué)和物理(理化)致癌因素誘發(fā)的腫瘤抗原

Experimentalinductionofimmunityagainsttumorcellsinducedbypolyomavirus(PV)病毒誘發(fā)的腫瘤抗原癌基因和突變型抑癌基因表達(dá)的腫瘤抗原在不同致癌因素和特定條件下，原癌基因可被異常激活，抑癌基因發(fā)生突變，導(dǎo)致正常細(xì)胞癌變，并能檢測(cè)到異常表達(dá)產(chǎn)物或相應(yīng)的編碼蛋白。這類蛋白在胞內(nèi)經(jīng)處理分解為不同小肽，通過MHCI類分子遞呈可作為腫瘤特異性抗原肽被T細(xì)胞所識(shí)別，激活CTL反響。Wild-typep53isrequiredtorestraincellgrowth.Itsactivitymaybelostbydeletionofbothwild-typeallelesorbyadominantmutationinoneallele.突變的抑癌基因編碼蛋白

Activationofagrowthfactorreceptorinvolvesligandbinding,dimerization,andautophosphorylation,Atruncatedoncogenicreceptorthatlackstheligand-bindingregionisconstitutivelyactivebecauseitisnotrepressedbytheN-terminaldomain.癌基因異常激活蛋白PathwaysthatrelyonRascouldfunctionbycontrollingeitherGNRForGAP.OncogenicRasmutantsarerefractorytocontrol,becauseRasremainsintheactiveform.

Translocationsbetweenchromosome22andchromosome9generatePhiladephiachromosomesthatsynthesizebcl-ablfusiontranscriptsthatareresponsiblefortwotypesofleukemia.

機(jī)體中某些染色體易位形成新的癌基因，所編碼蛋白由于融合點(diǎn)上出現(xiàn)新的氨基酸順序和形成新的空間構(gòu)象，僅在惡性細(xì)胞中表達(dá)，并與細(xì)胞的惡性轉(zhuǎn)化密切相關(guān),是T細(xì)胞識(shí)別的特異性腫瘤抗原。染色體易位產(chǎn)生的融合蛋白Alteration Functionofprotein TumortypePointmutationERBB2 Growthfactorreceptor BreastcarcinomaFMS CSF-1receptor AML,myelodysplasiaRas GTP-bindingprotein Carcinomasandothersp53 Tumorsuppressorcellcyclecontrol Manyincludingbladder, colon,lungRB1 Tumorsuppressorcellcyclecontrol Retinoblastoma, osteosarcoma PancreaticcarcinomaChromosomaltranslocationBCR-ABL Tyrosinekinase CML,ALLEZA-PRL Transcriptionfactor Pre-BcellALLH4-RET Growthfactorreceptor/tyrosinekinase ThyroidcarcinomaTPR-MET Growthfactorreceptor/tyrosinekinase GastriccarcinomaLMYC-RLF Transcriptionfactor SmallcelllungcarcinomaNPM/ALK Tyrosinekinase LymphomaDeletionmutations

ERB-B Growthfactorreceptor GlimasGeneticalterationsinhumantumorsproducingnewproteinsequences正常靜止基因表達(dá)的腫瘤抗原腫瘤細(xì)胞中某些被T細(xì)胞所識(shí)別的抗原往往由正常狀態(tài)下的靜止基因(silentgenes)所表達(dá)，除人的正常睪丸細(xì)胞外，這些基因一般只在惡性細(xì)胞中被激活而呈異常高表達(dá),又被稱為C-T抗原(cancer-testisantigen)；其中,MAGE家族至少有14個(gè)成員(MAGE-1～MAGE-14)，它們之間具有高度同源性(80～90％)；MAGE家族成員表達(dá)特征：在不同腫瘤中有不同程度的表達(dá)，多個(gè)成員也可在同一腫瘤中表達(dá)。同一腫瘤中可測(cè)出多種靜止基因的表達(dá)。這類基因的編碼蛋白經(jīng)胞質(zhì)溶膠途徑處理成小肽，通過MHCI類分子遞呈于腫瘤細(xì)胞外表，被CD8+T細(xì)胞所識(shí)別。TumourantigensrecognizedbyTlymphocytes:atthecoreofcancerimmunotherapy.NATUREREVIEWS|CANCERVOLUME14|FEBRUARY2021

ImmunogenicityofNY-ESO-1,MAGE-A1,MAGE-A3,andSSX-2CTantigenmRNAExpressionfrequencyFrequencyofserum T-cellepitopesantibodyincancerpatients CD8

CD4NY-ESO-134%Melanoma9%MelanomaA2DR425%OvarianCancer12%OvarianCancerA3116%LungCancer4%LungCancerCw3 24%BreastCancer 8%BreastCancer Cw6MAGE-A116%Melanoma 1%MelanomaA1B35Cw2DR13 28%OvarianCancer 3%OvarianCancerA3B7Cw3DR15 49%LungCancer 4%LungCancer A24B53Cw16 18%BreastCancer 0BreastCancer A28MAGE-A336%Melanoma 2%Melanoma A1B35DR4 17%OvarianCancer 0OvarianCancer A2B37DR7 47%LungCancer 0LungCancer A2B40DR11 11%BreastCancer 0BreastCancer A24B44 DR13 A24B52SSX-235%Melanoma 1%Melanoma A2 Unknown 0OvarianCancer 0OvarianCancer 17%LungCancer 0LungCancer 7%BreastCancer 0BreastCancerImmunologicalReviews2002;Vol188:22–32mRNAMAGE-1蛋白309氨基酸HLA-Cw16HLA-A1161169EADPTGHSYSAYGEPRKL

開放閱讀框230238MAGE-1基因X染色體q28區(qū)

外顯子3

外顯子2

外顯子1MAGE-1.Cw16肽段

MAGE1基因定位于X染色體q28區(qū)，mRNA轉(zhuǎn)錄表達(dá)的MAGE-1蛋白為309氨基酸，由HLA-A1和HLA-Cw16提呈的抗原肽位于161～169和230～238區(qū)域氨基酸序列。MAGE-1基因、編碼蛋白和抗原肽

腫瘤相關(guān)性抗原胚胎性抗原分化抗原癌基因高表達(dá)的抗原異常表達(dá)的糖脂/糖蛋白抗原ElevationofAFPandCEAinserumofpatientswithvariousdiseases胚胎性抗原細(xì)胞發(fā)生惡性轉(zhuǎn)化時(shí)，胚胎抗原編碼基因可被激活呈異常表達(dá)，出現(xiàn)在細(xì)胞質(zhì)、膜外表或分泌在血流中，其蛋白含量與細(xì)胞的惡性程度往往呈正相關(guān)。此類抗原一般難以激發(fā)機(jī)體產(chǎn)生抗體，但發(fā)現(xiàn)某些抗原可經(jīng)胞質(zhì)溶膠途徑處理成抗原肽由MHCI類分子遞呈于細(xì)胞膜外表，被T細(xì)胞識(shí)別。分化抗原分化抗原是細(xì)胞在分化成熟不同階段出現(xiàn)的抗原，不同來源、不同分化階段的細(xì)胞可表達(dá)不同的分化抗原。這些抗原在多種黑色素瘤細(xì)胞呈異常高表達(dá)、結(jié)構(gòu)高度同源，即很少顯示個(gè)體差異。異常表達(dá)的分化抗原可經(jīng)胞內(nèi)途徑處理成為抗原肽，通過MHCI或II類分子遞呈于細(xì)胞外表，被CD8+或CD4+T細(xì)胞所識(shí)別。某些腫瘤細(xì)胞癌基因表達(dá)產(chǎn)物未發(fā)現(xiàn)突變特征僅有表達(dá)量的差異。這些過度表達(dá)蛋白中某些肽經(jīng)MHCI類分子遞呈于細(xì)胞外表可被機(jī)體CD8+T細(xì)胞所識(shí)別，同時(shí)在某些患者體內(nèi)可測(cè)出相應(yīng)的抗體。癌基因高表達(dá)的抗原黑色素瘤特異性CTL識(shí)別的黑色素細(xì)胞分化抗原肽分化抗原 抗原肽結(jié)構(gòu) 肽位置 遞呈分子酪氨酸酶

MLLAVLYCL

1-9

HLA-A2

YMNGTMSQV

369-377

HLA-A2

AFLPWHRLF(L)

－

HLA-A24

SEIWRDIDF

192-200

HLA-B44Pmel17/gp100

KTWGQYWQV

154-162

HLA-A2

ITDQVQGSV

209-217

HLA-A2

YLEPGPVTA

280-288

HLA-A2

LLGDTATLRL

457-466

HLA-A2

VLYRYGSFSV

476-485

HLA-A2Melan-AMART-1

(E)AAGIGILTV

26(7)-35

HLA-A2

ILTVILGVL

32-40

HLA-A2gp75TRP1

－

－

HLA-A31Overexpressionofoncogene-encodedproteinsinhumantumorsProtein Normalfunction TumorknowntoexpresshighlevelsoftheproteinCyclinD1 RegulatoroftheG1-S Breastcancer(approx.20%),Coloncancer(20%) transitionviadependentkinase Thyroidcancer,Livercancer(11-13%)CyclinE RegulatoroftheG1-Stransition Breastcancer(90%) viabindtocyclin-dependentkinaseMdm2 Nuclearphosphoprotein. Sarcomas(30-36%),Leukaemia(between42-73%) Inhibitsthefunctionofp53 breastcancercellline,Gliomas(10-15%) byinteractionwiththeproteinReceptortyrosinekinasesEGFreceptorReceptorfortheepidermal Breastcancer(30-40%),Lungcancer(80%),Gliomas, growthfactor Bladdercancer(70%),Renalcancer(73%),Headand neckcancer(50%)ormore,dependentontype)ErbB2 Receptorbelongingtothesame Breastcancer(26%),Ovariancancer(20-30%),Stomach(Her-2/neu)familyastheEGFRreceptor.Cancer(4%)Non-small-celllungcancer(upto30%).

Ligandnotclearlyidentified.NuclearoncogenesC-myc Transcriptionfactor Breastcancer(6-57%),Small-celllung(20-30%), Cervicalcancer(30%),Testicularcancer,Colon cancerandHeadandneckcancer.ProteinsinvolvedintheregulationofapoptosisBcl-2 inhibitsapoptosis Non-Hodgkin’slymphoma(40-80%)WTp53 Tumorsuppressorprotein, Gliomas(astrocytomas)(60-80%) transcriptionfactorcaninduce Headandneckcancer(34%),Sarcoma(17%),apoptosisinresponsetoDNAdamage.

Acutemyeloidleukaemia(69%) 異常表達(dá)的糖脂/糖蛋白抗原某些腫瘤細(xì)胞可出現(xiàn)膜結(jié)構(gòu)改變，表達(dá)過量或結(jié)構(gòu)異常的糖脂和糖蛋白，其中包括神經(jīng)節(jié)苷脂、血型抗原和粘蛋白等。外表結(jié)構(gòu)改變和異常有助于腫瘤的侵襲和轉(zhuǎn)移。這類異常的糖脂和糖蛋白可誘發(fā)B細(xì)胞產(chǎn)生抗體和激發(fā)CTL反響。應(yīng)用相應(yīng)單抗檢測(cè)其含量，可為腫瘤的診斷和預(yù)后判斷提供參考。

B細(xì)胞CD4+T細(xì)胞CD8+T細(xì)胞MHCI類分子MHCII類分子Th1細(xì)胞因子

T細(xì)胞抗原特異T前體細(xì)胞NK

LAK活化M

直接裂解Th2細(xì)胞因子B細(xì)胞腫瘤細(xì)胞ADCCCDC激活補(bǔ)體分子腫瘤細(xì)胞克隆增殖APCTCRpMHCTCRpMHC激活的CD8+CTL降解成8-10寡肽降解成14-25寡肽CD4+T腫瘤細(xì)胞

CD8+T腫瘤抗原誘導(dǎo)凋亡機(jī)體抗腫瘤的免疫機(jī)制

MechanismofImmuneResponsetoTumor一、體液免疫抗腫瘤作用二、細(xì)胞免疫抗腫瘤作用

免疫系統(tǒng)識(shí)別腫瘤細(xì)胞表達(dá)抗原產(chǎn)生免疫應(yīng)答，激活效應(yīng)細(xì)胞和釋放一系列效應(yīng)分子，攻擊和去除腫瘤細(xì)胞。一、體液免疫抗腫瘤作用Tumor二、細(xì)胞免疫抗腫瘤作用1.T細(xì)胞CD4+T細(xì)胞與CD8+T細(xì)胞3.NK細(xì)胞4.巨噬細(xì)胞CD4+T細(xì)胞TumorCathepsins(blue)LFA-1(green)Talin(red)CD8+T細(xì)胞

殺傷腫瘤細(xì)胞過程分為效靶細(xì)胞結(jié)合，攻擊殺傷和靶細(xì)胞裂解。顯示對(duì)腫瘤細(xì)胞效應(yīng)功能具有高度的特異性和有效性。NK細(xì)胞特點(diǎn)NK細(xì)胞是一類對(duì)多種靶細(xì)胞自發(fā)性細(xì)胞毒活性的淋巴細(xì)胞譜系的特殊亞群，不表達(dá)T、B細(xì)胞特有外表標(biāo)志物(TCR、BCR、CD4和CD8等)，人類NK細(xì)胞表達(dá)CD16和CD56等分化抗原，占外周淋巴細(xì)胞的10-15%。殺傷靶細(xì)胞的機(jī)制

抗體依賴的細(xì)胞介導(dǎo)的細(xì)胞毒作用受體介導(dǎo)殺傷作用受體介導(dǎo)殺傷作用（MHCI類分子

）KIR和/或CD94/NKG2活化受體活化受體配體正常細(xì)胞NK細(xì)胞MHCI類分子－＋腫瘤細(xì)胞NK細(xì)胞殺傷＋受體介導(dǎo)的NK細(xì)胞激活及其對(duì)腫瘤細(xì)胞的殺傷機(jī)制

MHCI類分子是KIR配體，相互作用產(chǎn)生抑制性信號(hào)，可抑制NK細(xì)胞激活。當(dāng)腫瘤細(xì)胞I類分子表達(dá)下降或缺陷，缺乏炎癥信號(hào)，抑制NK細(xì)胞激活，發(fā)揮殺傷效應(yīng)。抗體依賴的細(xì)胞介導(dǎo)的細(xì)胞毒作用(ADCC)巨噬細(xì)胞巨噬細(xì)胞參與非特異性免疫和特異性免疫，殺傷腫瘤細(xì)胞機(jī)制主要為：①吞噬和殺傷作用；②介導(dǎo)炎癥反響；③釋放細(xì)胞因子參與免疫調(diào)節(jié)；④加工遞呈抗原、啟動(dòng)免疫應(yīng)答；⑤參與ADCC釋放效應(yīng)分子殺傷靶細(xì)胞。一、腫瘤細(xì)胞免疫原性低下二、免疫增強(qiáng)三、效應(yīng)細(xì)胞的功能異常四、腫瘤微環(huán)境中的免疫抑制細(xì)胞和相關(guān)分子腫瘤的免疫逃逸機(jī)制

MechanismofTumorEvasionoftheImmuneSystem一、腫瘤細(xì)胞免疫原性低下1.抗原表達(dá)不穩(wěn)定性2.MHCI類分子表達(dá)異常3.腫瘤細(xì)胞抗原加工處理缺陷4.缺乏共刺激信號(hào)Lowimmunogenicityandantigenicmodulation遺傳不穩(wěn)定性和抗原表達(dá)異質(zhì)性抗原調(diào)變

抗原表達(dá)不穩(wěn)定性NatureRev.CancerVol.15,473-483(2021)MHCI類分子表達(dá)異常腫瘤細(xì)胞外表MHCⅠ類分子表達(dá)下降或喪失2微球蛋白表達(dá)異常A1A33B8B44Cw3Cw4ABCw3A1B8CCw3A1A3Cw4Cw3B8Cw4B44DCw3A1Cw4B8E腫瘤細(xì)胞異常的HLA表型A.正常細(xì)胞HLAI類分子表型(舉例)。腫瘤細(xì)胞異常HLAI類分子表型：B.全部喪失，包括HLA-A、B、C；C.HLA一條單元型喪失，即HLA-A、B、三位點(diǎn)均喪失一半等位基因；D.某一位點(diǎn)喪失(此處為HLA-B或A位點(diǎn));E.某一等位基因喪失(此處為HLA-B44)。參見表18-5CD8+CTL內(nèi)源性抗原蛋白酶體TAPER抑制蛋白酶體活性：EBV，人CMV阻斷MHC合成和ER潴留：腺病毒，人CMV阻斷TAP轉(zhuǎn)運(yùn)：HSV從ER中移除MHCI類分子：CMVMHCI類相關(guān)抗原加工提呈途徑病毒I類樣分子干擾CTL的識(shí)別：小鼠CMV抗原肽pMHC

某些病毒在腫瘤細(xì)胞內(nèi)可抑制蛋白酶體活性，阻斷TAP轉(zhuǎn)運(yùn)，干擾MHC分子合成及其與抗原肽的結(jié)合等，從而影響MHC分子-抗原肽復(fù)合物在腫瘤細(xì)胞膜上的表達(dá)和CD8CTL對(duì)腫瘤細(xì)胞的識(shí)別。腫瘤細(xì)胞抗原加工處理途徑的缺陷多種病毒干擾腫瘤細(xì)胞抗原加工處理途徑參見表18-6缺乏共刺激信號(hào)CD8T細(xì)胞腫瘤細(xì)胞CD28TCRMHCI類分子抗原肽CD28B7TCRMHCI類分子抗原肽T細(xì)胞增殖細(xì)胞毒效應(yīng)ABB7cDNACTL殺傷協(xié)同刺激信號(hào)〔B7分子〕與T細(xì)胞增殖、活化〔A〕腫瘤細(xì)胞缺乏B7分子，不能有效激活T細(xì)胞；〔B〕轉(zhuǎn)染B7基因的腫瘤細(xì)胞表達(dá)B7分子后可激活CTL顯示對(duì)腫瘤細(xì)胞的細(xì)胞毒活性。二、免疫增強(qiáng)在實(shí)驗(yàn)中發(fā)現(xiàn)，給荷瘤動(dòng)物輸入抗腫瘤免疫血清可促進(jìn)腫瘤細(xì)胞的生長，稱之為免疫增強(qiáng)?！蚕魅鯔C(jī)體的抗腫瘤能力，從而有利于腫瘤細(xì)胞逃避效應(yīng)細(xì)胞的識(shí)別和攻擊〕?，F(xiàn)認(rèn)為，免疫增強(qiáng)是由于血清中存在封閉因子(blockingfactor)，后者遮蓋了腫瘤細(xì)胞外表的抗原決定簇。腫瘤的免疫增強(qiáng)不僅與封閉因子有關(guān)，也可能涉及某些淋巴細(xì)胞。在過繼性免疫治療中發(fā)現(xiàn)，某些致敏淋巴細(xì)胞過繼性注入帶瘤宿主，對(duì)腫瘤細(xì)胞出現(xiàn)刺激而非抑制其生長。這些淋巴細(xì)胞有可能屬于抑制性細(xì)胞。三、效應(yīng)細(xì)胞的功能異常1.T細(xì)胞信號(hào)轉(zhuǎn)導(dǎo)途徑缺陷2.細(xì)胞因子產(chǎn)生異常引起Th1/Th2細(xì)胞漂移AbnormalanddeficientofTCR-mediatedsignaltransductionT細(xì)胞信號(hào)轉(zhuǎn)導(dǎo)途徑缺陷參見表18-7TwosubsetsofCD4+Tcellseachregulationbyproducingcytokines細(xì)胞因子產(chǎn)生異常引起Th1/Th2細(xì)胞漂移髓樣抑制性細(xì)胞M2巨噬細(xì)胞調(diào)節(jié)性T細(xì)胞腫瘤細(xì)胞分泌免疫抑制因子腫瘤微環(huán)境中存在多種蛋白酶和相關(guān)分子四、腫瘤微環(huán)境中的免疫抑制細(xì)胞和相關(guān)分子VEGF,GM-CSF,

腫瘤細(xì)胞腫瘤微環(huán)境DC成熟T細(xì)胞IL-10IL-12下降NK細(xì)胞精氨酸酶ROS

髓樣抑制細(xì)胞在腫瘤微環(huán)境中VEGF等因子誘導(dǎo)作用下，髓樣抑制細(xì)胞〔MSC〕通過分泌精氨酸酶、ROS和IL-10，下調(diào)效應(yīng)T細(xì)胞的殺傷活性,抑制DC細(xì)胞成熟。IL-10也可抑制巨噬細(xì)胞分泌IL-12，從而降低NK細(xì)胞的活性。髓樣抑制細(xì)胞發(fā)揮免疫抑制作用的可能機(jī)制髓樣抑制性細(xì)胞巨噬細(xì)胞TGF-

,IL-6,PGE2,COX-2M2巨噬細(xì)胞腫瘤抗原腫瘤細(xì)胞PGE2CCL22H鐵蛋白腫瘤浸潤巨噬細(xì)胞Treg耐受型DC效應(yīng)性T細(xì)胞CCR4CCR4CCR7CCR7CD45RA未致敏Treg記憶性Treg誘導(dǎo)耐受誘導(dǎo)Treg擴(kuò)增吸引Treg腫瘤細(xì)胞以及腫瘤浸潤的巨噬細(xì)胞分泌CCL22及H鐵蛋白，招募初始的CCR4+Treg細(xì)胞在腫瘤部位積聚，Treg細(xì)胞與耐受性DC細(xì)胞相互作用下，抑制效應(yīng)T細(xì)胞的功能。Treg細(xì)胞的誘導(dǎo)和免疫抑制作用調(diào)節(jié)性T細(xì)胞IL-10FasLFasIL-6

腫瘤細(xì)胞阻止凋亡

＋mTNFRsTNF-αBPsTNF-αTGF-

凋亡

效應(yīng)細(xì)胞腫瘤細(xì)胞產(chǎn)生的免疫抑制因子和表達(dá)蛋白對(duì)效應(yīng)細(xì)胞的作用--腫瘤細(xì)胞分泌免疫抑制因子FasLandthepossiblefatesofantitumorCTLs

ARG是L-精氨酸代謝通路中的關(guān)鍵酶之一，能催化L-精氨酸轉(zhuǎn)化成L-鳥氨酸，從而降低L-精氨酸的水平。而L-精氨酸缺乏可下調(diào)T細(xì)胞CD3

鏈的表達(dá)；L-鳥氨酸是體內(nèi)多胺生物合成的關(guān)鍵成分，而多胺是重要的內(nèi)源性腫瘤刺激因子，在腫瘤細(xì)胞轉(zhuǎn)化和腫瘤增殖中發(fā)揮重要作用。

L-精氨酸是NOS的作用底物，NOS能催化L-精氨酸產(chǎn)生NO和L-瓜氨酸。NOS2及其產(chǎn)物NO可以阻斷IL-2的信號(hào)轉(zhuǎn)導(dǎo)途徑，從而抑制T細(xì)胞增殖活化。ARG1和NOS可協(xié)同作用導(dǎo)致過氧化亞硝酸鹽ONOO－的產(chǎn)生，抑制T細(xì)胞功能。

COX-2過表達(dá)可刺激血管生成因子如VEGF和bFGF分泌，促進(jìn)血管形成；COX-2過表達(dá)能夠增加Bcl-2蛋白表達(dá)，抑制腫瘤細(xì)胞凋亡;COX-2能促進(jìn)Th2型免疫應(yīng)答;增強(qiáng)CD4+CD25+Treg的抑制功能，并誘導(dǎo)CD4+CD25－T表達(dá)foxp3。腫瘤微環(huán)境中存在多種蛋白酶和相關(guān)分子腫瘤的免疫治療〔Tumorimmunotherapy〕一、腫瘤的主動(dòng)免疫治療二、腫瘤的被動(dòng)免疫治療

免疫治療分為主動(dòng)免疫療法和被動(dòng)免疫療法兩大類。前者著重激發(fā)機(jī)體抗腫瘤免疫應(yīng)答能力；后者向宿主轉(zhuǎn)移有抗腫瘤活性的治療因子或細(xì)胞，抑制腫瘤生長。1.非特異性主動(dòng)免疫療法2.特異性主動(dòng)免疫療法腫瘤的主動(dòng)免疫治療非特異性刺激因子非特異性地激發(fā)機(jī)體的免疫系統(tǒng)，增強(qiáng)抗腫瘤免疫應(yīng)答能力，而到達(dá)殺傷腫瘤細(xì)胞的目的。目前常用的有卡介苗(BCG)、短小棒狀桿菌(PV)、左旋咪唑(Levamisole，LMS)和寡聚脫氧核苷酸(CpG)等。非特異性主動(dòng)免疫療法細(xì)胞因子特異性主動(dòng)免疫療法處理的瘤細(xì)胞作為疫苗腫瘤抗原(肽)和人工合成肽抗原作為疫苗Enhancementoftumorcellimmunogenicitybytransfectionofcostimulatorandcytokinegenes.TumorcellsthatdonotadequatelystimulateTcellsontransplantationintoananimalwillnotberejectedandwillthereforegrowintotumors.Vaccinationwithtumorcellstransfectedwithgenesencodingcostimulatorsorcytokines,suchasIL-2,canleadtoenhancedactivationofTcells.Thisapproachofusingtransfectedtumorcellsasvaccineshasworkedinmousemodels,butclinicaltrialshavenotyetbeensuccessful.Tumorvaccines.Twotypesoftumorvaccinesthathaveshownefficacyinclinicaltrialsandanimalmodelsareillustrated.Autologousdendriticcellsarepreparedfrompatients’ownperipheralbloodcells.Thedendriticcellsareeitherpulsedwithrecombinantproteinortransfectedwithageneconstructthatexpressestheprotein.Theconstructmayalsoexpresscostimulatorymolecules(notshown).基因工程疫苗UseofDNAvaccinesraisesbothhumoralandcellularimmunityDNA疫苗DC治療性疫苗DC是專職抗原提呈細(xì)胞，負(fù)責(zé)對(duì)抗原進(jìn)行加工處理后提呈給T細(xì)胞，誘導(dǎo)T細(xì)胞的活化和增殖，激發(fā)有效的免疫應(yīng)答。由目前基于DC的治療性腫瘤疫苗，主要分為兩種，一是DC體外別離培養(yǎng)、擴(kuò)增鑒定后，荷載特定抗原回輸?shù)交颊唧w內(nèi)，二是誘導(dǎo)DC在體內(nèi)增殖、成熟，有效攝取特定腫瘤抗原提呈并激活T細(xì)胞。二者的共同目的都是最大程度活化腫瘤抗原特異性CD4T和CD8T細(xì)胞，發(fā)揮抗腫瘤效應(yīng)。目前認(rèn)為，單純DC疫苗雖有療效但仍須改進(jìn)，需要尋找更為適宜的腫瘤抗原、更有效地促進(jìn)DC增殖活化的方法或者與其他方法聯(lián)合使用。需在整體設(shè)計(jì)實(shí)施合理，以保證在促進(jìn)DC抗腫瘤效應(yīng)，同時(shí)注意檢測(cè)負(fù)向調(diào)節(jié)方面的變化。AppropriateutilizationandregulationofDCsinvaccinedesigninduceamuchmorepotentCTLantitumorimmuneresponse.(a)Tumorantigen-loadingtechniquesactivateDCsexvivo.(b)TargeteddrugsfacilitatethecaptureoftumorantigensbyDCsandtheexpressionofcostimulatorymoleculesandMHC-IIinvivo.(c)StimulatoryadjuvantsinducematurationofDCsandenhancetheactivationofCTLs.Aneffectivevaccineagainsthumanpapillomavirus(HPV)inducesantibodiesthatprotectagainstHPVinfection.Serotype16ofHPV(HPV-16)ishighlyassociatedwiththedevelopmentofcervicalcancer.Inaclinicaltrial,755healthyuninfectedwomenwereimmunizedwithavaccinegeneratedfromhighlypurifiednoninfectious'virus-likeparticles'(VLP)consistingofthecapsidproteinL1ofHPV-16andformulatedwithanalumadjuvant(inthiscasealuminumhydroxyphosphatesulfate).Incomparisonwiththeverylowtitersofantibodyinplacebo-treateduninfectedwomen(greenline),orwomenpreviouslyinfectedwithHPVthatreceivedplacebo(blueline),thewomentreatedwiththevirus-likeparticlevaccine(redline)developedhightitersofantibodyagainsttheL1capsidprotein.NoneoftheseimmunizedwomensubsequentlybecameinfectedbyHPV-16.Ananti-HPVvaccinemarketedasGardasilisnowavailableandrecommendedforuseingirlsandyoungwomenasaprotectionfromcervicalcancercausedbyHPVserotypes6,11,16,and18.腫瘤的被動(dòng)免疫治療

1.過繼性免疫療法

2.抗體導(dǎo)向療法過繼性免疫療法過繼性免疫療法是指把自身或異體的具有抗腫瘤活性的免疫血清或免疫細(xì)胞轉(zhuǎn)輸?shù)矫庖吖δ艿拖碌哪[瘤患者，在體內(nèi)發(fā)揮抗腫瘤作用，以此到達(dá)治療腫瘤的目的。淋巴因子激活的殺傷細(xì)胞〔LAK〕腫瘤浸潤性淋巴細(xì)胞〔TIL〕LAK細(xì)胞的制備和應(yīng)用腫瘤患者

全血細(xì)胞（淋巴細(xì)胞）加IL-2，培養(yǎng)、擴(kuò)增過繼性輸注抗腫瘤淋巴細(xì)胞加IL-2培養(yǎng)、擴(kuò)增的淋巴細(xì)胞Before

AfterTreatmentofmelanomawithLAKcellsandIL-2淋巴因子激活的殺傷細(xì)胞小鼠脾臟細(xì)胞或人外周血淋巴細(xì)胞在體外培養(yǎng)中，經(jīng)高濃度的細(xì)胞因子〔主要為IL-2〕誘導(dǎo)后發(fā)生擴(kuò)增，產(chǎn)生一類能非特異性地殺傷自身和異體腫瘤細(xì)胞的效應(yīng)細(xì)胞，稱為淋巴因子激活的殺傷細(xì)胞〔lymphokineactivatedkillercell〕,簡(jiǎn)稱LAK細(xì)胞。

LAK細(xì)胞是一群異質(zhì)性的細(xì)胞群，主要來源于外周血淋巴細(xì)胞，其表型既可是CD3＋細(xì)胞，也可是CD3－細(xì)胞，往往具有NK細(xì)胞樣標(biāo)記〔CD16和CD56〕，其殺傷腫瘤細(xì)胞不需要抗原致敏，亦無MHC約束性。TIL細(xì)胞的制備和應(yīng)用腫瘤患者腫瘤切除

腫瘤浸潤淋巴細(xì)胞(TIL)加IL-2反復(fù)培養(yǎng)ELISA測(cè)刺激后IFN-

水平用IL-2及抗CD3單抗克隆擴(kuò)增TIL過繼性輸注抗腫瘤淋巴細(xì)胞陽性克隆治療前以化療作非髓系淋巴細(xì)胞清除BeforeAfterTreatmentofMelanomaswithTIL+IL-2腫瘤浸潤性淋巴細(xì)胞〔tumor-infiltrationlymphocytes,TIL〕嵌合抗原受體(CAT)Examplesoftumorantigensthathavebeentargetedbymonoclonalantibodiesintherapeutictrials.抗體導(dǎo)向療法人源化抗體前藥活性藥物腫瘤細(xì)胞酶mAb-核素結(jié)合物mAb-蓖麻毒素結(jié)合物雙特異性抗體（抗腫瘤/抗CD16）NK細(xì)胞人源性Fc段嵌合抗體免疫細(xì)胞因子（IL-2）腫瘤細(xì)胞雙特異性抗體（抗腫瘤/抗CD3）T細(xì)胞CD16CD3A．制備抗腫瘤的抗體與效應(yīng)分子〔毒蛋白、化療藥物或放射性同位素〕偶聯(lián)物，利用抗體特異性識(shí)別腫瘤抗原作用，使效應(yīng)分子有效地到達(dá)腫瘤部位，選擇性殺傷腫瘤細(xì)胞。B.雙特異性抗體中包含著兩種不同識(shí)別特異性抗原的Fab段，通過特異結(jié)合腫瘤抗原同時(shí)結(jié)合不同效應(yīng)細(xì)胞和分子，到達(dá)有效殺傷腫瘤的作用?？贵w偶聯(lián)物/雙特異性抗體的抗腫瘤作用機(jī)制雙特異性抗體抗體偶聯(lián)物Mechanismsoftumourcellkillingbyantibodies.a|Directtumourcellkillingcanbeelicitedbyreceptoragonistactivity,suchasanantibodybindingtoatumourcellsurfacereceptorandactivatingit,leadingtoapoptosis(representedbythemitochondrion).Itcanalsobemediatedbyreceptorantagonistactivity,suchasanantibodybindingtoacellsurfacereceptorandblockingdimerization,kinaseactivationanddownstreamsignalling,leadingtoreducedproliferationandapoptosis.Anantibodybindingtoanenzymecanleadtoneutralization,signallingabrogationandcelldeath,andconjugatedantibodiescanbeusedtodeliverapayload(suchasadrug,toxin,smallinterferingRNAorradioisotope)toatumourcell.b|Immune-mediatedtumourcellkillingcanbecarriedoutbytheinductionofphagocytosis;complementactivation;antibody-dependentcellularcytotoxicity(ADCC);geneticallymodifiedTcellsbeingtargetedtothetumourbysingle-chainvariablefragment(scFv);Tcellsbeingactivatedbyantibody-mediatedcross-presentationofantigentodendriticcells;andinhibitionofTcellinhibitoryreceptors,suchascytotoxicTlymphocyte-associatedantigen4(CTLA4).c|Vascularandstromalcellablationcanbeinducedbyvasculaturereceptorantagonismorligandtrapping(notshown);stromalcellinhibition;deliveryofatoxintostromalcells;anddeliveryofatoxintothevasculature.MAC,membraneattackcomplex;MHC,majorhistocompatibilitycomplex;NK,naturalkiller.Antibodytherapyofcancer.NATUREREVIEWS.CANCERVOLUME12APRIL2021Strategiestoinactivateand/ordepleteCD4+CD25+Tregcellsincancer.BasedonthecurrentlyavailableevidenceinfavourofTreg-mediatedsuppressionofanti-tumorT-cellresponses,severalstrategiestoinactivateand/ordepleteTregshavebeenevaluatedinpre-clinicalmodelsofcancer.ChemotherapeuticdrugssuchasCYandfludarabineinterferewithTregfunctions.AbdirectedagainstAgassignedtoTregcells(CD25,GITR,CTLA-4,CD4)depleteand/orfunctionallyinhibitTregs,thuscontributingtotherapeuticefficacyofconcomitantlyadminis