1、细胞免疫抗肿瘤作用细胞免疫抗肿瘤作用体液免疫抗肿瘤作用体液免疫抗肿瘤作用细胞因子抗肿瘤的作用细胞因子抗肿瘤的作用Tumour antigens recognized by T lymphocytes:at the core of cancer immunotherapy.NATURE REVIEWS|CANCER VOLUME 14|FEBRUARY 2014 细胞免疫抗肿瘤作用细胞免疫抗肿瘤作用一、一、T细胞细胞二、二、NK细胞细胞三、三、NKT细胞细胞四、巨噬细胞四、巨噬细胞(M)T细胞在抗肿瘤的特异性细胞在抗肿瘤的特异性免疫中是主要效应细胞。免疫中是主要效应细胞。1 1.T.T细胞的分化
2、、成熟和活化细胞的分化、成熟和活化2 2.T.T细胞抗肿瘤的效应机制细胞抗肿瘤的效应机制一、一、T细胞细胞T T细胞的分化、成熟和活化细胞的分化、成熟和活化 T T细胞在胸腺中分化、成熟细胞在胸腺中分化、成熟 T T细胞抗原受体和相关分子细胞抗原受体和相关分子 抗原的识别和抗原的识别和T T细胞活化、增殖细胞活化、增殖Most of the immature thymocytes in the thymus die either because they make an unproductive TCR-gene rearrangement or because they fail posit
3、ive or negative selection.Three mature T-cell populations(green,pink,blue)are produced and move to the peripheral lymphoid organs.Most T cells express the TCR and either CD4(blue)or CD8(pink).A few T cells express the TCR(green);most of these lack both CD4 and CD8.Note that the boundary between the
4、early and the late phase of T-cell development is not sharp.T T细胞在胸腺中分化、成熟细胞在胸腺中分化、成熟 Positive and negative selection of thymocytes in the thymus.Thymic selection involves thymic stromal cells(epithelial cells,dendritic cells,and macrophages),and results in mature T cells that are both self-MHC rest
5、ricted and self-tolerant.Positive and negative selection of thymocytes in the thymus阳性选择阳性选择（positive selection）在在胸腺皮质中，胸腺皮质中，CD4CD4+CD8CD8+双阳性双阳性T T细胞，其细胞，其TCRTCR能与胸腺基质细胞能与胸腺基质细胞表面的表面的MHCMHC/类分子类分子-抗原肽结合，且具抗原肽结合，且具适当（低）亲和力适当（低）亲和力的的DPDP细细胞分化为单阳性（胞分化为单阳性（SPSP）T T细胞，其中与细胞，其中与类分子结合的类分子结合的DPDP细胞分化为细胞分化
6、为CD8CD8+T T细胞（细胞（SPSP）；与）；与类分子结合的类分子结合的DPDP细胞分化为细胞分化为CD4CD4+T T细胞（细胞（SPSP）；）；而不能与而不能与MHC-MHC-抗原肽结合抗原肽结合或高亲和力的或高亲和力的DPDP细胞则发生凋亡遭克隆清细胞则发生凋亡遭克隆清除。此过程也称为胸腺的阳性选择除。此过程也称为胸腺的阳性选择。生物学。生物学意义：意义：赋予成熟的赋予成熟的T T细胞细胞具有具有MHCMHC限制性限制性。经历阳性选择的经历阳性选择的SPSP细胞在胸腺的皮髓质交界处及髓质区还须经细胞在胸腺的皮髓质交界处及髓质区还须经历阴性选择：凡是能识别自身抗原历阴性选择：凡是能识
7、别自身抗原-MHC-MHC复合物、且具有高亲和力的复合物、且具有高亲和力的SPSP细胞发生凋亡遭克隆清除，其实质是清除自身反应性细胞发生凋亡遭克隆清除，其实质是清除自身反应性T T细胞，即阴细胞，即阴性选择。生物学意义：性选择。生物学意义：赋予成熟的赋予成熟的T T细胞具有自身免疫耐受（中枢）细胞具有自身免疫耐受（中枢）的特性的特性。T T细胞经三个发育阶段及胸腺选择后成为成熟细胞经三个发育阶段及胸腺选择后成为成熟T T细胞，迁出胸腺细胞，迁出胸腺进入外周进入外周T T细胞库。细胞库。阴性选择阴性选择（negative selection）T T细胞抗原受体和相关分子细胞抗原受体和相关分子 H
8、oming of lymphocytes Examples of homing receptors and vascular addressins involved in selective trafficking of nave and effector T cells.(a)Nave T cell tend to home to secondary lymphoid tissue through their HEV regions.(b,c)Various subsets of effector T cells express high levels of particular homin
9、g receptors that allow them to home to endothelium in particular tertiary extralymphoid tissues.抗原的识别和抗原的识别和T T细胞活化、增殖细胞活化、增殖膜表面分子膜表面分子相互作用相互作用 Overview of TCR-mediated signaling.TCR engagement by peptide-MHC complexes initiates the assembly of a signaling complex.An early step is the Lck-mediated p
10、hosphorylation of ITAMs on the zeta chains of the TCR complex,creating docking sites to which the protein kinase ZAP-70 attaches and becomes activated by phosphorylation.A series of ZAP-70-catalyzed protein phosphorylations enable the generation of a variety of signals.信号转导信号转导效应功能效应功能CD4CD4+T T细胞细胞
11、CD8CD8+T T细胞细胞 +T T细胞细胞TregTreg细胞细胞T T细胞抗肿瘤的效应机制细胞抗肿瘤的效应机制CD4CD4+T T细胞细胞CD8CD8+T T细胞细胞增殖分化增殖分化杀伤效应杀伤效应 杀伤肿瘤细胞过程分为效靶细胞结合，杀伤肿瘤细胞过程分为效靶细胞结合，攻击杀伤和靶细胞裂解。显示对肿瘤细胞攻击杀伤和靶细胞裂解。显示对肿瘤细胞效应功能具有高度的特异性和有效性。效应功能具有高度的特异性和有效性。Stages in CTL-mediated killing of target cells Comparison of morphologic changes that occur i
12、n apoptosis and necrosis.Apoptosis,which results in the programmed cell death of hematopoietic cells,does not induce a local inflammatory response.In contrast,necrosis,the process that leads to death of injured cells,results in release of the cells contents,which may induce a local inflammatory resp
13、onse.Apoptosis.Light micrographs of (a)normal thymocytes(developing T cells in the thymus)and(b)apoptotic thymocytes.Scanning electron micrographs of(c)normal and(d)apoptotic thymocytes.The Cancer-Immunity Cycle.The generation of immunity to cancer is a cyclic process that can be self propagating,le
14、ading to an accumulation of immune-stimulatory factors that in principle should amplify and broaden T cell responses.The cycle is also characterized by inhibitory factors that lead to immune regulatory feedback mechanisms,which can halt the development or limit the immunity.This cycle can be divided
15、 into seven major steps,starting with the release of antigens from the cancer cell and ending with the killing of cancer cells.Each step is described above,with the primary cell types involved and the anatomic location of the activity listed.Abbreviations are as follows:APCs,antigen presenting cells
16、;CTLs,cytotoxic T lymphocytes.Oncology Meets Immunology:The Cancer-Immunity Cycle.Immunity 39,July 25,2013CD4 调节性调节性 T 细胞的分化及功能特点细胞的分化及功能特点自然调节T细胞（CD25+Foxp3+nTreg）直接分化于胸腺，通过细胞间接触以CTLA-4等行使抑制功能。适应性调节T细胞（CD25+Foxp3+iTreg）在TGF-及IL-2诱导下自nTreg或Foxp3-T细胞产生，借助分泌IL-10和TGF-发挥抑制作用。另外两种调节T细胞（Tr1和Th3）分化自CD25-F
17、oxp3-T细胞，其中Tr1的产生由多种因素（IL-10,Vit D3,imDC）参与诱导，通过分泌IL-10和TGF-发挥作用。免免疫疫抑抑 制制 CTLA-4IL-10,TGF-Foxp3+nTreg Th3Tr1 Foxp3+iTreg胸腺TGF-TGF-IL-10等 自身肽-MHC II亲和力()自身肽-MHC II亲和力()CD25+Foxp3+TCD25Foxp3TIL-10,TGF-起源 细胞前体 调控因素 Treg 亚群 效应分子 功能Treg细胞细胞 +T T细胞细胞Time course of appearance of thymocytes and thymocytes
18、during mouse fetal development.The graph shows the percentage of CD3+cells in the thymus that are double-negative(CD48)and bear the T-cell receptor(black)or are double positive(CD4+8+)and bear the T-cell receptor(blue).+T T细胞是指细胞是指TCRTCR和和链构成的链构成的T T细细胞，在胞，在T T细胞中占细胞中占5%5%。通常表达。通常表达CD3CD3和和CD56CD56分
19、子，大多数不表达分子，大多数不表达CD4CD4和和CD8CD8分子。分子。该该TCRTCR直接识别磷酸抗原和某些单一肽类分子配体。直接识别磷酸抗原和某些单一肽类分子配体。在抗肿瘤作用中，主要通过分泌多种细胞因子，如在抗肿瘤作用中，主要通过分泌多种细胞因子，如IL-IL-2 2、IL-4IL-4、IFN-IFN-和和TNF-TNF-等，其作用类似于等，其作用类似于CD4CD4+T T细胞；细胞；其效应作用可释放胞内的细胞毒颗粒杀伤靶细胞，其其效应作用可释放胞内的细胞毒颗粒杀伤靶细胞，其作用类似于作用类似于CD8CD8+T T细胞，但杀伤作用不受细胞，但杀伤作用不受MHCMHC限制性。限制性。Tu
20、mor cell ligands recognized by human T cells and strategies for T cellbased immunotherapy.Left panel,the dominant population of V2V9 T cells recognizes via its TCR nonpeptidic phosphoantigens,such as the naturally occurring isoprenoid metabolite IPP,the synthetic bromohydrin pyrophosphate(BrHPP;Phos
21、phostim),aminobisphosphonates(N-BP),the ectopically expressed F1-ATPase,and apolipoprotein A-I.Ligands recognized by the V1 TCR have been less well defined but include MICA.In addition,MICA/MICB and ULBPs frequently expressed on tumor cells are ligands for the activating NK receptor NKG2D,which is p
22、resent on V1 and V2 T cells.Right panel,strategies for T cellbased immunotherapy include(i)the adoptive cell transfer of in vitro expanded T cells and(ii)the in vivo activation of V2V 9 T cells by phosphoantigens(e.g.,bromohydrin pyrophosphate/Phosphostim)or aminobisphosphonates and low-dose IL-2.V2
23、V9 T cells can be readily activated and expanded in vitro by phosphoantigens(bromohydrin pyrophosphate)or aminobisphosphonates in the presence of IL-2.Possible beneficial effects of additional NKG2D stimulation by activating antibodies or naturally occurring NKG2D ligands require further investigati
24、on.In the absence of defined ligands(except for MICA)that are recognized by the V1 TCR,tumor-reactive V1 T cells can be activated and expanded in vitro by stimulation with mitogenic anti-TCR antibodies and IL-2 in the absence or presence of additional NKG2D ligation.Ligands recognized by human T cel
25、lStrategies for T cell-based immunotherapyTCRNKG2DNKG2DTCRAPO-A-1F1-ATPaseIPP,BrHPP,NBPMICA/BULBPsULBPsMICA/BV 1V 2Adoptive cell transferIn vivo activation+BrHPP or NBP+IL-2+/-NKG2DBrHPP or NBP+IL-2+anti-TCR antibody+IL-2+NKG2DV 1V 2Cancer Res 2007;(67):1,5-8二、二、NKNK细胞细胞 NK细胞是抗肿瘤细胞是抗肿瘤免疫早期起重要作用的免疫早期
26、起重要作用的效应细胞，参与非特异效应细胞，参与非特异性免疫和特异性免疫。性免疫和特异性免疫。主要通过受体介导主要通过受体介导杀伤杀伤作用和抗体依赖的细胞作用和抗体依赖的细胞介导的细胞毒作用。介导的细胞毒作用。1.NK1.NK细胞的特性细胞的特性2.NK2.NK细胞的效应功能细胞的效应功能NKNK细胞特性细胞特性 NK细胞是一类对多种靶细胞自发性细胞毒活性的淋巴细细胞是一类对多种靶细胞自发性细胞毒活性的淋巴细胞谱系的特殊亚群，不表达胞谱系的特殊亚群，不表达T、B细胞特有表面标志物（细胞特有表面标志物（TCR、BCR、CD4和和CD8等），人类等），人类NK细胞表达细胞表达CD16和和CD56等分
27、等分化抗原，占外周淋巴细胞的化抗原，占外周淋巴细胞的10-15%。效应功能：ADCCLAK天然细胞毒CD56brightIL-2c-kitCCR7 CD16dimL-selectinhighNKR：KIRCD94-NKG2A高分泌细胞因子：GM-CSF,TNF-/-IFN-,IL-10CD94-NKG2ACD16brightKIRNKR：KIRCD94-NKG2ACD56dim IL-2RCXCR1CX3CR1 效应功能：ADCC LAK 天然细胞毒 低分泌 细胞因子 PEN5-PSGL-1CD56brightNK和和CD56dimNK细胞亚群的特征细胞亚群的特征识别识别HLA 类分子的抑制或
28、活化受体类分子的抑制或活化受体杀伤细胞免疫球蛋白样受体杀伤细胞免疫球蛋白样受体(KIR)(KIR)杀伤细胞凝集素样受体杀伤细胞凝集素样受体(KLR)(KLR)识别非识别非HLA 类分子的活化受体类分子的活化受体NKG2DNKG2D自然细胞毒性受体自然细胞毒性受体(NCR(NCR)NKNK细胞杀伤活性受体细胞杀伤活性受体和抑制受体和抑制受体Recognition of tumour cells by NK cells.a|Natural killer(NK)cells are tolerant to healthy host cells,as the strength of the activa
29、ting signals they receive on encountering these cells is dampened by the engagement of inhibitory receptors(tolerance).b|Tumour cells may lose expression of MHC class I molecules.NK cells become activated in response to these cells,as they are no longer held in check by the inhibitory signal deliver
30、ed by MHC class I molecule engagement.This is known as missing-self triggering of NK cell activation.c|In addition,NK cells are selectively activated by stressed cells,which upregulate activating ligands for NK cells and thereby overcome the inhibitory signalling delivered by MHC class I molecules.T
31、his is known as stress-induced self triggering of NK cell activation.In both conditions,NK cell activation leads to tumour elimination directly(through NK cell-mediated cytotoxicity)or indirectly(through the production of pro-inflammatory cytokines,such as interferon).Targeting natural killer cells
32、and natural killer T cells in cancer.NATURE REVIEWS|IMMUNOLOGY VOLUME 12|APRIL 2012 NKNK细胞识别模式细胞识别模式（19861986年年Karre等）等）NK NK 细胞细胞的的“丢失自我丢失自我”和和“诱导自我诱导自我”识别模式识别模式A.早期NK细胞与表达MHC I类分子的正常细胞（教育细胞）相互作用，获得辨别自身正常细胞的能力（称为受教育），当此NK细胞接受IL-15等多种细胞因子刺激后进一步激活（被致敏）。B.NK细胞活化性受体及抑制性受体间的平衡，决定NK细胞是否激活并对靶细胞实施杀伤。右上：健康细
33、胞 I 类分子与NK细胞抑制性受体结合，产生的抑制信号抑制活化性受体活性，NK细胞不杀伤靶细胞（耐受状态）。右中和右下：受到应急诱导的细胞（肿瘤或病毒感染细胞）I 类分子表达下调（“丢失自我”识别模式），或活化性配体表达上调（“诱导自我”识别模式），抑制信号不能发挥作用，导致NK对靶细胞的杀伤。受教育MHC I类分子MHC I类专一抑制性受体IL-12IL-12RIL-18IL-18RIL-2,IL-15IL-2R,IL-15R(链+c)IL-21IL-21RIFN-IFN-R-活化受体活化配体耐受状态 丢失自我诱导自我 被致敏教育细胞NK 健康细胞应急细胞NKNKNK -应急细胞NKABMH
34、C I类分子MHC I类专一抑制性受体DNA损伤,肿瘤恶变,病毒感染NK细胞IFN-,TNF-等细胞因子M树突状细胞应急诱导性靶细胞ADCCT细胞靶细胞裂解清除应急诱导性靶细胞抗原提呈 正常细胞 抗原提呈 NK细胞的细胞的免疫生物学免疫生物学功能功能NK细胞可以识别并以抗体依赖细胞介导细胞毒（ADCC）形式杀伤多种应急诱导的靶细胞(如DNA损伤细胞、肿瘤细胞和病毒感染细胞等），NK细胞还可以促进DC细胞成熟和发挥抗原提呈功能，参与激活T细胞。另外，NK细胞可以通过分泌IFN-,TNF-或IL-10的方式来提高或者降低巨噬细胞或树突状细胞的活性。NKNK细胞的效应功能细胞的效应功能杀伤作用和杀伤
35、作用和免疫调节免疫调节诱导活化产生IFN-/，TNF-,IL-12NK细胞在抗病毒中的作用细胞在抗病毒中的作用 小鼠实验表明，病毒感染后体内最早出现的是I型干扰素、TNF-和IL-12，以及由NK细胞参与的杀伤，然后是 病毒特异性细胞毒性T细胞（CTL）发挥作用，病毒滴度下降。三、三、NKT细胞细胞CD1dDN-TDP-TSP-TNK TTCR-NKT前体CD4+CD8+MHCIIMHC IMHC IMHC II凋亡凋亡 自然杀伤自然杀伤T T细胞细胞(nature killer T cells,NKT)是一群细胞是一群细胞表面具有表面具有TCRTCR又表达又表达NKNK细胞受体细胞受体的淋巴
36、细胞亚群。的淋巴细胞亚群。NKTNKT细胞的分化发育细胞的分化发育A.主流前体模式:NKT细胞与传统T细胞一样在胸腺中发育，由CD4+CD8+双阳性(DP)T细胞经CD1d分子的选择，逐渐脱离主流分化途径，获得NK细胞表型，最终形成NKT细胞，进入外周血、肝脏和脾脏中。B.特定前体模式：NKT细胞通过非胸腺依赖的途径发育分化，即从可能存在的NKT前体细胞独立地在外周器官(如肝脏)中分化成熟。DN-T：双阴性(CD4-CD8-)T细胞；SP-T：单阳性(CD4+或CD8+)T细胞。NKT细胞识别由细胞表面CD1分子递呈的脂类抗原。NKT细胞识别抗原不同于T细胞,但它们的激活和T细胞一样需要共刺激
37、信号。NKT细胞（经典）的TCR识别DC的CD1递呈的-半乳糖神经酰胺(-galactosylceramide,-GalCer),同 时,其 表 面 的 C D 2 8 及CD154(CD40L)分别与DC表面的CD80/CD86(B7-1/B7-2)及CD40相互作用,在激活DC产生IL-12及IL-18同时使NKT细胞表面的IL-12R与IL-18R表达上调。NKT细胞被IL-12与IL-18进一步激活,产生高水平的IFN-与IL-4。由脾脏等外周NKT细胞受到刺激后主要产生IFN-。NKTNKT细胞对抗原的识别细胞对抗原的识别 Cellular and molecular mechani
38、sms of antitumor immune responses mediated by-GalCer-activated NKT cells.The-GalCer/CD1d complex activates NKT cells to upregulate CD40L and cytotoxic molecule expressions.CD40L on NKT cells stimulates CD40 on DC,leading to their activation to produce IL-12.IL-12 from DC activates NKT cells to produ
39、ce IFN-which in turn stimulates NK cells and CD8 CTLs mediating antitumor cytotoxicity.-GalCer-activated NKT cells also induce maturation of DC,which contributes to the upregulation of Th1 responses.免疫调节免疫调节：NKTNKT细胞激活后，可通过分泌细胞激活后，可通过分泌IL-4IL-4和和IFN-IFN-调节调节ThTh细胞的分化细胞的分化,对免疫应答和自身免疫的调节。对免疫应答和自身免疫的调节
40、。-半乳糖神经酰胺 Natural killer T(NKT)cells recognize glycolipids bound to CD1d at the surface of antigen-presenting cells(APCs).Activated NKT cells secrete IFN-in a CD40-and IL-12-dependent manner and can lyse tumor cells though an MHC-unrestricted,TRAIL or perforin-dependent pathway.细胞毒作用细胞毒作用：NKT细胞被活化后，
41、可分化成具有细胞毒细胞被活化后，可分化成具有细胞毒性和杀伤活性的效应细胞性和杀伤活性的效应细胞,在免疫、保护性免疫应答中在免疫、保护性免疫应答中发挥抗肿瘤作用。发挥抗肿瘤作用。NKTNKT细胞效应功能细胞效应功能Antitumour activities of iNKT cells.a|Tumour cells that express CD1d can be directly recognized by invariant natural killer T(iNKT)cells and subsequently eliminated either directly,by iNKT cell
42、activity,or indirectly via iNKT cell-mediated activation of natural killer(NK)cells.b|Although not all tumour cells express CD1d,iNKT cells can also become activated in response to CD1d expressing antigen-presenting cells(APCs).The activated iNKT cells then promote NK cell activation and thereby ind
43、irectly mediate tumour cell elimination.c|It has also been proposed that iNKT cells can limit tumour growth by suppressing the production of pro-angiogenic factors by macrophages,although this still remains to be confirmed.TAM,tumour-associated macrophage;TCR,T cell receptor.Targeting natural killer
44、 cells and natural killer T cells in cancer.NATURE REVIEWS|IMMUNOLOGY VOLUME 12|APRIL 2012 四、单核四、单核/巨噬细胞巨噬细胞1.1.来源和来源和组织分布组织分布2.2.表面标志表面标志3.3.生物学功能和调节生物学功能和调节 单核单核/巨巨噬细胞噬细胞的来源和的来源和组织分布组织分布单核单核巨巨噬细胞噬细胞表面标志表面标志葡聚糖受体 甘露糖受体 Toll样受体 CR3 LPS受体(CD14)清道夫受体 B7分子 MHCI类分子 TNF-CD40 NO O2 TNF受体 MHC II 类分子 AB巨噬细胞
45、表达的受体和表面分子巨噬细胞表达的受体和表面分子 A.A.巨噬细胞；巨噬细胞；B.B.活化的巨噬细胞活化的巨噬细胞 巨噬细胞参与固有免疫和特异性免巨噬细胞参与固有免疫和特异性免疫，杀伤肿瘤细胞机制主要为：疫，杀伤肿瘤细胞机制主要为：吞噬和杀伤作用；吞噬和杀伤作用；介导炎症反应；介导炎症反应；加工递呈抗原、启动免疫应答；加工递呈抗原、启动免疫应答；释放细胞因子参与免疫调节；释放细胞因子参与免疫调节；参与参与ADCC释放效应分子杀伤靶细胞。释放效应分子杀伤靶细胞。生物学功能和调节生物学功能和调节 Mediators of antimicrobial and cytotoxic activity o
46、f macrophages and neutrophils体液免疫抗肿瘤作用体液免疫抗肿瘤作用 体液免疫是指抗体所发挥的效应功能。体液免疫是指抗体所发挥的效应功能。一、一、B细胞的分化、成熟和活化、增殖细胞的分化、成熟和活化、增殖二、生发中心的形成、体细胞突变和抗体类别转换二、生发中心的形成、体细胞突变和抗体类别转换三、三、B细胞抗原受体和信号转导细胞抗原受体和信号转导四、体液免疫抗肿瘤效应机制四、体液免疫抗肿瘤效应机制一、一、B细胞的分化、成熟和活化、增殖细胞的分化、成熟和活化、增殖 Overview of B-cell development.During the antigen-inde
47、pendent maturation phase,immuno-competent B cells expressing membrane IgM and IgD are generated in the bone marrow.Only about 10%of the potential B cells reach maturity and exit the bone marrow.Naive B cells in the periphery die within a few days unless they encounter soluble protein antigen and act
48、ivated TH cells.Once activated,B cells proliferate within secondary lymphoid organs.Those bearing high-affinity mIg differentiate into plasma cells and memory B cells,which may express different isotypes because of class switching.The numbers cited refer to B-cell development in the mouse,but the ov
49、erall principles apply to humans as well.二、生发中心的形成、体细胞突变和抗体类别转换二、生发中心的形成、体细胞突变和抗体类别转换Germinal centers are sites of intense cell proliferation and cell death.The photomicrograph(left panel)shows a high-power view of a section through a human tonsillar germinal center.Closely packed centro blasts seen
50、 in the lower part of this photomicrograph form the so-called dark zone of the germinal center.Above this region is the less densely packed light zone.The right panel shows immunofluorescent staining of a germinal center.B cells are found in the dark zone,light zone,and mantle zone.Proliferating cel
