肿瘤与免疫-肿瘤免疫治疗-课件

肿瘤免疫肿瘤免疫治疗治疗 Tumor immunotherapy概概 述述主动免疫疗法主动免疫疗法 被动免疫疗法被动免疫疗法肿瘤免疫治疗 概 述1Paul Ehrlichs magic bullet concept:100 years of progress,Nature Reviews/Cancer Vol.8,2008概概 述述Paul Ehrlichs magic bullet co2NATURE REVIEW/CANCER,2008,Vol.8:299-307NATURE REVIEW/CANCER,2008,Vol3 肿肿瘤瘤免免疫疫治治疗疗分分为为主主动动免免疫疫疗疗法法和和被被动动免免疫疫疗疗法法两两大大类类。前前者者着着重重激激发发机机体体抗抗肿肿瘤瘤免免疫疫应应答答能能力力；后后者者向向宿宿主主转移有抗肿瘤活性的治疗因子或细胞，抑制肿瘤生长。转移有抗肿瘤活性的治疗因子或细胞，抑制肿瘤生长。肿瘤免疫治疗分为主动免疫疗法和被动免疫疗法两大类。前4 肿瘤免疫疗法肿瘤免疫疗法的分类及常用的生物制剂的分类及常用的生物制剂 分类分类治疗治疗因子或细胞因子或细胞 主动免疫疗法主动免疫疗法非特非特异性异性 BCG、CpG ODN、HSP、IFN-等等 特异性特异性 减减毒或灭活的瘤苗毒或灭活的瘤苗、修饰、修饰的瘤苗的瘤苗、肿瘤抗原肿瘤抗原肽肽疫苗和基因重组产物疫苗和基因重组产物 被动免疫被动免疫疗法疗法非特异性非特异性细胞因子诱导的杀伤细胞细胞因子诱导的杀伤细胞（CIK）淋巴因子激活的杀伤细胞淋巴因子激活的杀伤细胞（LAK）DCDC介导的免疫治疗介导的免疫治疗 特异性特异性肿瘤浸润淋巴细胞肿瘤浸润淋巴细胞（TIL)抗体抗体（Antibody）免疫偶免疫偶联联物物（Immuno-conjugates）嵌合抗原受体（嵌合抗原受体（CAT）BCG:Bacille Calmette-Guerin,CpG ODN:CpG Oligodeoxynucleotide,HSP:heat shock protein,CIK:cytokine inducing killer cell,LAK:lymphokine activated killer cell,TIL:tumor-infiltration lymphocytes,CAT：chimeric antigen receptor 肿瘤免疫疗法的分类及常用的生物制剂5Approved immune therapies for cancerApproved immune therapies for 6Antibody therapy of cancer.NATURE REVIEWS.CANCER VOLUME 12 APRIL 2012 ADCC,antibody-dependent cellular cytotoxicity;CDC,complement-dependent cytotoxicity;CLL,chronic lymphocytic leukaemia;CTLA4,cytotoxic T lymphocyte-associated antigen 4;EGFR,epidermal growth factor receptor;FDA,US Food and Drug Administration;IgG,immunoglobulin G;INFa;interferon-a;NHL,non-Hodgkins lymphoma;NSCLC,non-small-cell lung cancer;SCCHN,squamous cell carcinoma of the head and neck;VEGF,vascular endothelial growth factor.*Based on information from the European Medicines Agency.Not recommended for patients with colorectal cancer whose tumours express mutated KRAS.Antibody therapy of cancer.NA7一、非特异性主动免疫疗法一、非特异性主动免疫疗法二、特异性主动免疫疗法二、特异性主动免疫疗法主动免疫治疗主动免疫治疗(Active Immunotherapy)一、非特异性主动免疫疗法主动免疫治疗8 非特异性地激发机体的免疫系统，增强抗肿瘤免疫应非特异性地激发机体的免疫系统，增强抗肿瘤免疫应答能力，而达到杀伤肿瘤答能力，而达到杀伤肿瘤细胞。细胞。1 1.免疫因子免疫因子 胸腺素(Thymosin)，转移因子(Transter factor)免疫核糖核酸(Immune RNA)细胞因子:IFN-,IFN-,IFN-,IL-2,TNF-等2.2.微生物制剂微生物制剂 卡介苗(BCG)，CpG寡聚脱氧核苷酸(CpG ODN)3.3.化学合成药物化学合成药物 左旋咪唑(Levamisole，LMS)4.4.中药制剂中药制剂(滋阴、补气、补血)人参、黄芪等(主要为多糖成分)一、非特异性主动免疫疗法一、非特异性主动免疫疗法 非特异性地激发机体的免疫系统，增强抗肿瘤免疫应一、非9卡介苗卡介苗制剂制剂 牛型结核杆菌或其细胞壁成分或细胞骨架成分，牛型结核杆菌或其细胞壁成分或细胞骨架成分，BCG中的有效成分是中的有效成分是胞壁酰二肽胞壁酰二肽(muramyl dipeptide，MDP)。作用作用机制机制 直接直接活化活化M，刺激，刺激M 表达表达IL-6、IL-1,增强增强IFN-对对 M的刺激作用。的刺激作用。刺激刺激特异性特异性TDTH的产生，间接活化的产生，间接活化M。刺激刺激NK细胞增殖。细胞增殖。非特非特异性刺激单核巨噬细胞。异性刺激单核巨噬细胞。促进促进IL-2和和IL-4对对B细胞的作用。细胞的作用。临床临床应用应用 黑色素瘤黑色素瘤：瘤灶内直接注射：瘤灶内直接注射 浅浅表性膀胱癌：膀胱内滴注表性膀胱癌：膀胱内滴注卡介苗10卡介苗的临床应用卡介苗的临床应用（黑色素瘤）（黑色素瘤）皮肤转移灶内皮肤转移灶内BCGBCG直接注射可使直接注射可使60%60%左右接受注射的病灶消退，而且有左右接受注射的病灶消退，而且有15%15%未接受注射的病灶也消退，提示局部注射可导致全身性抗肿瘤免疫的未接受注射的病灶也消退，提示局部注射可导致全身性抗肿瘤免疫的建立。总共包括建立。总共包括269269名患者的名患者的1616项使用这一方法的研究都获得不同程度的项使用这一方法的研究都获得不同程度的疗效，完全缓解率最高达疗效，完全缓解率最高达90%(90%(范围范围7-90%)7-90%)，部分缓解率范围为，部分缓解率范围为5-50%5-50%。在一项结合放疗的试验中，在一项结合放疗的试验中，74%74%患者获得完全缓解，另外患者获得完全缓解，另外5%5%患者获得患者获得部分缓解。部分缓解。BCGBCG瘤内注射疗法可提高患者生存率。不用瘤内注射疗法可提高患者生存率。不用BCGBCG治疗的皮肤癌复发患者，治疗的皮肤癌复发患者，存活时间的中值为存活时间的中值为13.313.3月，而接受月，而接受BCGBCG瘤内注射患者的瘤内注射患者的5 5年生存率达年生存率达27%27%。有相当一部分原发性黑色素瘤患者经皮内注射有相当一部分原发性黑色素瘤患者经皮内注射BCGBCG治疗后，长期存活。治疗后，长期存活。卡介苗的临床应用卡介苗的临床应用（浅表性膀胱癌）（浅表性膀胱癌）BCG BCG膀胱内滴注可以消除肉眼可见的浅表性膀胱癌，并能预防复发。膀胱内滴注可以消除肉眼可见的浅表性膀胱癌，并能预防复发。膀膀胱胱内内注注射射BCGBCG用用于于预预防防膀膀胱胱癌癌复复发发，可可以以显显著著延延迟迟疾疾病病的的进进展展，延延长长保保留留膀膀胱胱的的时时间间，提提高高总总存存活活率率。5 5项项追追踪踪期期范范围围为为12-6012-60月月的的随随机机研研究究发发现现，具具有有高高度度复复发发危危险险的的患患者者，用用膀膀胱胱内内注注射射BCGBCG治治疗疗者者，70%70%保保持持无无瘤瘤；而用反复尿道内切除者，仅有而用反复尿道内切除者，仅有31%31%的患者保持无瘤。的患者保持无瘤。报道实例报道实例卡介苗的临床应用（黑色素瘤）皮肤转移灶内BCG11CpGCpG寡聚脱氧核苷酸寡聚脱氧核苷酸CpG寡聚脱氧核苷酸12肿瘤与免疫-肿瘤免疫治疗-课件13Activation of innate and adaptive immunity by TLR9 activation.Among human immune cells,only B cells and pDCs constitutively express TLR9.These cells endocytose DNA into an endosomal compartment where it binds to TLR9,forming a signaling complex.If the DNA contains unmethylated CpG motifs,TLR9 is stimulated,and the cell becomes activated.In pDCs,this results in type I IFN secretion,which activates NK cells,monocytes,and other APCs,and in the pDC maturation into a more effective APC able to activate naive T cells.Opposing these immune boosting effects,pDCs activated through TLR9 also mediate immune-suppressive effects through counter regulatory factors such as indoleamine 2,3-dioxygenase(35,36)and the generation of Tregs.In B cells,TLR9 stimulation results in the secretion of proinflammatory cytokines,such as IL-6,and in the release of immune regulatory cytokines that might limit the intensity of the inflammatory response,such as IL-10.TLR9 activation of B cells confers a greatly increased sensitivity to antigen stimulation and enhances their differentiation into antibody-secreting plasma cells.On balance,these immune effects of CpG DNA generally promote strong Th1 CD4+and CD8+T cell responses.However,the concurrent activation of counter regulatory pathways such as the induction of Tregs limit TLR9-induced immune activation,offering a potential for enhancing the therapeutic efficacy of TLR9 agonists by coadministration of antagonists of one or more of these inhibitory pathways.Activation of innate and adapt14肿瘤与免疫-肿瘤免疫治疗-课件15肿瘤与免疫-肿瘤免疫治疗-课件16细胞因子细胞因子Function of cytokines in host defense细胞因子Function of cytokines in h17 上调免疫细胞的表面分子和受体的上调免疫细胞的表面分子和受体的表达；表达；促进促进DCDC细胞的成熟细胞的成熟,增强增强T T细胞的细胞的增殖、分化和增殖、分化和CTLCTL的活的活 化和效应功能，化和效应功能，刺激刺激B B细胞产生抗体，提高细胞产生抗体，提高NKNK细胞活性，细胞活性，激发巨噬细胞等产生抗肿瘤免疫应答；激发巨噬细胞等产生抗肿瘤免疫应答；促进免疫效应细胞释放淋巴毒素和效应分子杀伤肿瘤；促进免疫效应细胞释放淋巴毒素和效应分子杀伤肿瘤；促进肿瘤细胞表达促进肿瘤细胞表达MHCMHC分子，增强肿瘤细胞的免疫原性分子，增强肿瘤细胞的免疫原性 和对效应细胞的敏感性；和对效应细胞的敏感性；某些细胞因子具有直接破坏肿瘤细胞和促使其发生凋某些细胞因子具有直接破坏肿瘤细胞和促使其发生凋 亡的作用，如亡的作用，如TNFTNF。细胞因子杀伤肿瘤细胞的效应机制细胞因子杀伤肿瘤细胞的效应机制 上调免疫细胞的表面分子和受体的表达；细胞因子杀伤肿瘤细胞18肿瘤与免疫-肿瘤免疫治疗-课件19二、特异性主动免疫疗法二、特异性主动免疫疗法1.肿瘤疫苗肿瘤疫苗2.抗独特型抗体作为疫苗抗独特型抗体作为疫苗 二、特异性主动免疫疗法20肿瘤疫苗肿瘤疫苗使用使用肿瘤疫苗的原理肿瘤疫苗的原理肿瘤肿瘤疫苗分类疫苗分类肿瘤肿瘤疫苗治疗肿瘤疫苗治疗肿瘤的可能影响因素的可能影响因素肿瘤疫苗使用肿瘤疫苗的原理21使用肿瘤疫苗的原理使用肿瘤疫苗的原理肿肿瘤瘤细细胞胞表表达达肿肿瘤瘤抗抗原原并并被被机机体体免免疫疫系系统统所识别；所识别；肿肿瘤瘤疫疫苗苗刺刺激激主主动动特特异异性性抗抗肿肿瘤瘤应应答答免免疫疫，激发或加强宿主免疫以消灭肿瘤；激发或加强宿主免疫以消灭肿瘤；肿肿瘤瘤患患者者中中许许多多已已知知的的和和未未知知的的因因素素（肿肿瘤免疫原性）造成了免疫应答的失败。瘤免疫原性）造成了免疫应答的失败。使用肿瘤疫苗的原理肿瘤细胞表达肿瘤抗原并被机体免疫系统所识别22肿瘤与免疫-肿瘤免疫治疗-课件23肿瘤疫苗分类肿瘤疫苗分类根据根据应用目的分类应用目的分类 预防性预防性肿瘤疫苗肿瘤疫苗治疗治疗性肿瘤疫苗性肿瘤疫苗 根据根据肿瘤疫苗中肿瘤抗原的存在形式分类肿瘤疫苗中肿瘤抗原的存在形式分类细胞细胞疫苗疫苗 灭活肿瘤细胞、灭活肿瘤细胞、DC、DC/肿瘤融合细胞肿瘤融合细胞肿瘤抗原疫苗肿瘤抗原疫苗 肿瘤肿瘤细胞裂解物、肿瘤抗原、抗原肽细胞裂解物、肿瘤抗原、抗原肽基因工程基因工程疫苗疫苗 肿瘤抗原肿瘤抗原、细胞因子或协同刺激分子细胞因子或协同刺激分子等等 (基因重组产物基因重组产物)基因基因转染转染DC、肿瘤细胞等肿瘤细胞等DNADNA疫苗疫苗 肿瘤抗原基因质粒肿瘤抗原基因质粒肿瘤疫苗分类根据应用目的分类 24u肿瘤细胞肿瘤细胞疫苗疫苗uDC疫苗疫苗uDC/肿瘤融合细胞肿瘤融合细胞 细胞疫苗细胞疫苗肿瘤细胞疫苗 细胞疫苗25 肿瘤细胞疫苗肿瘤细胞疫苗 肿瘤细胞疫苗26DC疫苗疫苗特征特征DC疫苗特征27制备和应用制备和应用制备和应用28 DC/肿瘤融合细胞肿瘤融合细胞 适用于大多数肿瘤抗原还未鉴定的肿瘤适用于大多数肿瘤抗原还未鉴定的肿瘤。DC/肿瘤融合细胞29肿瘤抗原的来源和应用肿瘤抗原的来源和应用 来源来源 举举 例例癌基因产物癌基因产物 ras12密码子突变密码子突变:胰腺癌胰腺癌 BCR/abl重排产物重排产物:CML 静止基因产物静止基因产物 MAGE家族家族:黑色素瘤，乳腺癌黑色素瘤，乳腺癌病毒基因产物病毒基因产物 EBV:Burkett淋巴瘤，鼻咽癌淋巴瘤，鼻咽癌 HPV:宫颈癌宫颈癌 HBV:肝细胞癌肝细胞癌组织特异性蛋白组织特异性蛋白(分化抗原分化抗原)酪氨酸酶酪氨酸酶:黑色素瘤黑色素瘤突变的抑癌基因产物突变的抑癌基因产物 P53:多种肿瘤多种肿瘤抗独特型抗体抗独特型抗体(Anti-Id-Ab)TCR Id:T细胞淋巴瘤细胞淋巴瘤肿瘤抗原疫苗肿瘤抗原疫苗肿瘤抗原的来源和应用 来源 30mRNAMAGE-1蛋白309氨基酸HLA-Cw16HLA-A1161 169 EADPTGHSY SAYGEPRKL 开放阅读框 230 238MAGE-1基因X染色体q28区 外显子3 外显子2 外显子1MAGE-1.Cw16肽段 MAGE1MAGE1基因定位于基因定位于X X染色体染色体q28q28区，区，mRNAmRNA转录表达的转录表达的MAGE-1MAGE-1蛋白为蛋白为309309氨基酸，由氨基酸，由HLA-A1HLA-A1和和HLA-Cw16HLA-Cw16提呈的抗原肽位于提呈的抗原肽位于161161169169和和230230238238区域氨基酸序列。区域氨基酸序列。MAGE-1MAGE-1基因、编码蛋白和抗原肽基因、编码蛋白和抗原肽mRNAMAGE-1蛋白HLA-Cw16HLA-A1161 31黑色素瘤特异性黑色素瘤特异性CTLCTL识别的黑色素细胞分化抗原肽识别的黑色素细胞分化抗原肽分化抗原分化抗原抗原肽结构抗原肽结构肽位置肽位置递呈分子递呈分子酪氨酸酶酪氨酸酶MLLAVLYCL1-9HLA-A2 YMNGTMSQV369-377HLA-A2 AFLPWHRLF(L)HLA-A24 SEIWRDIDF192-200HLA-B44Pmel 17/gp100KTWGQYWQV154-162HLA-A2 ITDQVQGSV209-217HLA-A2 YLEPGPVTA280-288HLA-A2 LLGDTATLRL457-466HLA-A2 VLYRYGSFSV476-485HLA-A2Melan-AMART-1(E)AAGIGILTV26(7)-35HLA-A2 ILTVILGVL32-40HLA-A2gp75TRP1HLA-A31黑色素瘤特异性CTL识别的黑色素细胞分化抗原肽分化抗原 抗原32制备和应用制备和应用制备和应用33肿瘤与免疫-肿瘤免疫治疗-课件34An effective vaccine against human papilloma virus(HPV)induces antibodies that protect against HPV infection.Serotype 16 of HPV(HPV-16)is highly associated with the development of cervical cancer.In a clinical trial,755 healthy uninfected women were immunized with a vaccine generated from highly purified noninfectious virus-like particles(VLP)consisting of the capsid protein L1 of HPV-16 and formulated with an alum adjuvant(in this case aluminum hydroxyphosphate sulfate).In comparison with the very low titers of antibody in placebo-treated uninfected women(green line),or women previously infected with HPV that received placebo(blue line),the women treated with the virus-like particle vaccine(red line)developed high titers of antibody against the L1 capsid protein.None of these immunized women subsequently became infected by HPV-16.An anti-HPV vaccine marketed as Gardasil is now available and recommended for use in girls and young women as a protection from cervical cancer caused by HPV serotypes 6,11,16,and 18.An effective vaccine against h35PLG based 3D polymer scaffold vaccine to recruit and program immune cells to generate antitumor immunity.(a)Overall schematic of the vaccine.1.SEM image of a PLG 3D scaffold;scale bar:1000 mm.(b)Number of CD11c+DCs isolated from the scaffold on day 14 post implantation in response to 0,400 ng,3000 ng and 7000 ng of GM-CSF.(c)The number of CD11c+CCR7+DCs isolated from the scaffold loaded with PEI-ODN control,10 mg PEI-CpG-ODN,400 and 3000 ng GM-CSF and 400 and 3000 ng GM-CSF in combination with 10 mg PEI-CpG-ODN at day 7 after implantation.(d)A comparison of the survival of mice following prophylactic(left)and therapeutic(right)vaccination.In the prophylactic vaccination,mice were vaccinated with blank PLG scaffolds(Blank),antigen+100 mg CpG-ODN(Lys+100CpG),antigen+3000 ng GM-CSF+10 mg CpG-ODN(Lys+3000GM+10CpG),antigen+3000 ng GM-CSF+100 mg CpG-ODN(Lys+3000GM+100CpG),or irradiated,GM-CSF-transduced B16-F10 cells(cell based).Mice were challenged(day 0 on graphs)with 105 B16-F10 melanoma tumor cells and monitored for the onset of tumor occurrence.In the therapeutic vaccination,mice were inoculated with 5 X 105 B16-F10 cells and allowed to develop for 9 days and treated with blank PLG matrices(Blank),PLG matrices loaded with 3000 ng of GM-CSF and 100 mg of CpG-ODN(GM+CpG).Mice were also treated once(Vax,1;at day 9),or twice(Vax,2;at days 9 and 19)with PLG matrices incorporating GM-CSF,CpG-ODN,and tumor lysates(Vax).Mice were also treated with 5 X105 irradiated,GM-CSF-transduced B16-F10 cells.Panels(c)and(d left)reproduced with permission 19,Copyright 2009,Nature Publishing Group.Panels(b)and(d right)reproduced with permission 20,Copyright 2009,The American Association for the Advancement of Science.Materials based tumor immunotherapy vaccines.Current Opinion in Immunology 2013,25:238245PLG based 3D polymer scaffold 36 基因工程疫苗基因工程疫苗l修饰的肿瘤疫苗修饰的肿瘤疫苗 协同刺激分子、细胞因子基因修饰的协同刺激分子、细胞因子基因修饰的 肿瘤细胞肿瘤细胞 l重组病毒疫苗重组病毒疫苗 用已知肽的用已知肽的cDNA序列与灭活病毒重组序列与灭活病毒重组 产生的疫苗，可同时转入产生的疫苗，可同时转入MHC、B7等等 基因。基因。基因工程疫苗修饰的肿瘤疫苗37Enhancement of tumor cell immunogenicity by transfection of costimulator and cytokine genes.Tumor cells that do not adequately stimulate T cells on transplantation into an animal will not be rejected and will therefore grow into tumors.Vaccination with tumor cells transfected with genes encoding costimulators or cytokines,such as IL-2,can lead to enhanced activation of T cells.This approach of using transfected tumor cells as vaccines has worked in mouse models,but clinical trials have not yet been successful.Tumor vaccines.Two types of tumor vaccines that have shown efficacy in clinical trials and animal models are illustrated.Autologous dendritic cells are prepared from patients own peripheral blood cells.The dendritic cells are either pulsed with recombinant protein or transfected with a gene construct that expresses the protein.The construct may also express costimulatory molecules(not shown).Enhancement of tumor cell immu38肿瘤与免疫-肿瘤免疫治疗-课件39Use of DNA vaccines raises both humoral and cellular immunity DNA疫苗疫苗Use of DNA vaccines raises bot40 肿肿瘤瘤DNADNA疫疫苗苗由由肿肿瘤瘤抗抗原原基基因因与与质质粒粒重重组组后后形形成成，其其中中肿肿瘤瘤抗抗原原基基因因随随治治疗疗的的肿肿瘤瘤的的类类型型及及选选用用的的肿肿瘤瘤抗抗原原不不同同而而不不同同，质质粒粒则则是是作作为为肿肿瘤瘤抗抗原原基基因因的的载载体体。肿肿瘤瘤DNADNA疫疫苗苗中中常常用用的的质质粒粒有有pSV2pSV2、pRSVpRSV、pcDNA3.1pcDNA3.1和和pC1pC1等等，这这些些载载体体一一般般都都设设计计有有真真核核基基因因表表达达调调控控序序列列（如如增增强强子子和和启启动动子子等等）、供供目目的的基基因因插插入入的的多多克克隆隆位位点点、转转录录终终止止序序列列以以及及可可使使质质粒粒在大肠杆菌中保持并多拷贝复制的序列在大肠杆菌中保持并多拷贝复制的序列ColEIColEI等等。肿肿瘤瘤DNADNA疫疫苗苗的的免免疫疫接接种种有有多多种种途途径径，包包括括直直接接注注射射、基基因因枪枪免免疫疫和和电电穿穿孔孔免免疫疫等等。不不同同免免疫疫途途径径的的免免疫疫机机制制不不同同，所所诱诱导导的的免免疫疫保保护护力力强强弱弱和和维维持持时时间间也也不不尽尽相相同同。肌肌肉肉注注射射是最早采用的免疫途径，研究最充分，效果也较好。是最早采用的免疫途径，研究最充分，效果也较好。肿肿瘤瘤DNADNA疫疫苗苗的的临临床床应应用用目目前前报报道道较较少少，仅仅有有少少量量进进入入I/III/II期期临临床床试试验验，如如gp100gp100和和蛋蛋白白酪酪氨氨酸酸激激酶酶用用于于治治疗疗黑黑色色素素瘤瘤、CEACEA疫疫苗苗用用于于治治疗疗结结直直肠肠癌癌、PSAPSA和和PSMAPSMA用用于于治治疗疗前前列列腺腺癌癌、HPV-E6HPV-E6，E7E7用用于于治治疗疗子子宫宫颈颈癌癌以以及及NY-ESO-1NY-ESO-1用用于于治治疗疗非小细胞性肺癌等。非小细胞性肺癌等。肿瘤DNA疫苗由肿瘤抗原基因与质粒重组后形成41Appropriate utilization and regulation of DCs in vaccine design induce a much more potent CTL antitumor immune response.(a)Tumor antigen-loading techniques activate DCs ex vivo.(b)Targeted drugs facilitate the capture of tumor antigens by DCs and the expression of costimulatory molecules and MHC-II in vivo.(c)Stimulatory adjuvants induce maturation of DCs and enhance the activation of CTLs.Appropriate utilization and re42肿瘤疫苗治疗肿瘤的可能影响因素肿瘤疫苗治疗肿瘤的可能影响因素肿瘤抗原选择肿瘤抗原选择MHC I、II类分子和共刺激分子的缺陷或降低类分子和共刺激分子的缺陷或降低诱导诱导T细胞细胞CTLA-4的表达的表达CD4+CD25+Foxp3+调节调节T T细胞的抑制作用细胞的抑制作用TH1/TH2细胞不平衡细胞不平衡免疫程序优化组合免疫程序优化组合(1)(1)抗原和佐剂的剂量及途径应用恰当抗原和佐剂的剂量及途径应用恰当(2)(2)减轻肿瘤负荷减轻肿瘤负荷(3)(3)免疫活性细胞含量和功能检测免疫活性细胞含量和功能检测(4)(4)抑制性抑制性DCDC、TregTreg细胞、细胞、MSCMSC和抑制性巨噬细胞等含量分析和抑制性巨噬细胞等含量分析(5)(5)肿瘤治疗的干预手段联合应用肿瘤治疗的干预手段联合应用肿瘤疫苗治疗肿瘤的可能影响因素肿瘤抗原选择43 Schematic illustration of human telomerase reverse transcriptase(hTERT)antigenic peptides presented by MHC class I complex on the tumor cell surface.Multiple antigenic peptides derived from hTERT in tumor cell are recognized by immune cells.The peptides are generated by proteasome degradation of hTERT protein.The hTERT peptides are transported by the transporter associated with antigen processing into the ER where they are loaded onto MHC class I molecules and transported through the Golgi apparatus onto the cell surface.CD8+cytotoxic T lymphocyte with specific receptors recognize and attack the tumor cell by the engagement of the T cell receptor with an MHC class I-hTERT peptide complex.Biochimica et Biophysica Acta xxx(2009)Telomerase in cancer immunotherapy(review)Schematic illustration 44 Model of human telomerase reverse transcriptase(hTERT)antigen stimulation of DCs,CD4+T cells,and expansion of CD8+T cells in vivo.(1)Telomerase hTERT antigen transfer and processing in APC,(2)antigen presentation from APC or tumor cells to T cells,(3)interaction between CD4+and CD8+T cells,(4)Expansion of CD8+CTLs that have sufficient telomeres,(5)CTLs with short telomeres undergo cell senescence and apoptosis,and(6)CD8+cytotoxic lymphocytes attack the tumors expressing telomerase hTERT antigen.Model of human telomera45Hypothetical modes of action of dominant and cryptic immune epitopes in epitope-specific T cell clonal evasion or reactivation.(A)Dominant epitope to MHC class I forms a stable antigen-MHC class I complex that engages with its specific TCR at such high affinity that death of the T cell clone occurs.(B)An intermediate epitope with variable avidity to MHC class I forms antigen-MHC complexes that interact with its specific TCR discontinuously under particular conditions resulting in either elimination or activation of the T cell clone.(C)Cryptic epitope with low affinity to MHC class I forms an unstable antigen-MHC class I complex that has a poor interaction with its specific TCR,resulting in the survival and continuous presence of the T cell clone specific to the cryptic epitope.(D)Position 1 tyrosine substitution(P1Y)or position 9 valine substitution(P9V)in the cryptic epitope enhances the cryptic epitope interaction with MHC class I and thereby establishes an optimal complex to interact with its specific TCR,resulting in activation and expansion of the T cell clone.Activated T cell clones bind their specific antigen epitope-MHC class I complex on tumor cells to exert kill effects on the tumor cells.Biochimica et Biophysica Acta xxx(2009)Telomerase in cancer immunotherapy(review)Hypothetical modes of action o46Advantages and Disadvantages of Different Vaccination StrategiesAdvantages and Disadvantages o47Ways to improve the clinical outcome of peptide-based vaccinesWays to improve the clinical o48The repertoire of human tumor-associated epitopes-identification and selection of antigens and their application in clinical trials.Current Opinion in Immunology 2013,25:277283The repertoire of human tumor-49肿瘤与免疫-肿瘤免疫治疗-课件50激活质质控控癌症患者 DC 激发性DC抗原靶向引入途径剂量频率成熟耐受性DCDC亚群DC疫苗有效治疗肿瘤的临床和免疫学指疫苗有效治疗肿瘤的临床和免疫学指标进行质量控制和标准化运用标进行质量控制和标准化运用激活质癌症患者 DC 激发性D51AbAg免疫系统感受淋巴细胞克隆容积的阈值免疫系统感受淋巴细胞克隆容积的阈值TCR/BCR 储备库:51013 11018 多样性淋巴细胞抗原受体多样性的体内储备及其对抗原激发的感知和应答淋巴细胞抗原受体多样性的体内储备及其对抗原激发的感知和应答 免疫网络学说免疫网络学说 带有不同独特型抗原受体的淋巴细胞克隆，因数量未能超过免疫系统的感知阈值而在体内长期存在。图中以不同的几何图形代表各种细胞克隆的受体（TCR或BCR）。抗原（红三角）一旦到来，选择出带有相应受体的淋巴细胞克隆使之发生增殖并跨越阈值，成为Ab1（紫红色），后者再诱导产生Ab2（蓝色和绿色），引发网络式的连锁反应。注意Ab2作为抗原分子，其表位结构与抗原分子相似。19741974年年Niels Niels JerneJerne提出抗体分子上的独特型和抗独特型相互识别而形成免疫网络（免疫网络学说）。提出抗体分子上的独特型和抗独特型相互识别而形成免疫网络（免疫网络学说）。抗独特型抗体作为疫苗抗独特型抗体作为疫苗AbAg免疫系统感受淋巴细胞克隆容积的阈值TCR/BCR 储52Preparation and application of anti-idiotype antibodyPreparation and application of53一、过继性免疫疗法一、过继性免疫疗法二、抗体导向二、抗体导向疗法疗法三、其他相关免疫疗法三、其他相关免疫疗法被动免疫治疗被动免疫治疗 (Passive Immunotherapy)一、过继性免疫疗法被动免疫治疗 54 一一、过继性免疫疗法、过继性免疫疗法 过过继继性性免免疫疫疗疗法法(Adoptive immunotherapy)是是指指把把自自身身或或异异体体的的具具有有抗抗肿肿瘤瘤活活性性的的免免疫疫细细胞胞转转输输到到免免疫疫功功能能低低下下的的肿肿瘤瘤患患者者，在在体体内内发发挥挥抗抗肿瘤作用，以此达到治疗肿瘤的目的。肿瘤作用，以此达到治疗肿瘤的目的。1 1.淋巴因子激活的杀伤细胞淋巴因子激活的杀伤细胞(LAK)(LAK)2 2.肿瘤浸润性淋巴细胞肿瘤浸润性淋巴细胞(TIL)(TIL)一、过继性免疫疗法55 小鼠脾脏细胞或人外周血淋巴细胞在体外培养中，经高浓小鼠脾脏细胞或人外周血淋巴细胞在体外培养中，经高浓度的细胞因子度的细胞因子（IL-2IL-2）诱导后发生扩增，产生一类能）诱导后发生扩增，产生一类能非特非特异性异性杀伤杀伤自身和异体肿瘤细胞的效应细胞，称为自身和异体肿瘤细胞的效应细胞，称为淋巴因子激活的杀淋巴因子激活的杀伤细胞（伤细胞（lymphokine activated killer lymphokine activated killer cellcell）,简称简称LAKLAK细胞。细胞。淋巴因子激活的杀伤细胞淋巴因子激活的杀伤细胞 LAK LAK细胞的特征细胞的特征 是是一一群群异异质质性性的的细细胞胞群群，主主要要来来源源于于外外周周血血淋淋巴巴细细胞胞，其其表表型型既既可可是是CD3CD3细细胞胞，也也可可是是CD3CD3细细胞胞，往往往往具具有有NKNK细细胞胞样样标标记记（CD16CD16和和CD56)CD56)，其其杀杀伤伤肿肿瘤瘤细细胞胞不不需需要要抗抗原原致致敏敏，亦无亦无MHCMHC约束性。约束性。IL-2/LAKIL-2/LAK细胞抗肿瘤机制细胞抗肿瘤机制 LAKLAK细细胞胞对对肿肿瘤瘤细细胞胞的的非非特特异异性性直直接接杀杀伤伤；依依赖赖肿肿瘤瘤免免疫疫原原性性的的特特异异性性抗抗瘤瘤机机制制 对对具具有有免免疫疫原原性性肿肿瘤瘤有有效效;有有长期疗效，说明特异性免疫记忆的存在长期疗效，说明特异性免疫记忆的存在。小鼠脾脏细胞或人外周血淋巴细胞在体外培养中，经高浓度的56Cancer therapies targeting NK cells.a.Following haplo-identical or MHC-matched haematopoietic stem cell transplantation(HSCT),natural killer(NK)cells of donor origin develop in the patient with cancer.b.Alternatively,NK cell populations can be isolated from healthy donors and activated and/or expanded in vitro before infusion into the patient with cancer.In both cases(allogeneic HSCT and NK cell infusion),the aim is to promote the antitumour function of donor NK cells in the patient.Indeed,a fraction of donor NK cells will be not be inhibited by the MHC class I molecules of the patient,as the killer cell immunoglobulin-like receptors(KIRs)expressed by the donor NK cells will not interact with the MHC class I molecules of the patient,and this promotes tumour cell elimination.In contrast to cancer cells,most healthy cells of the patient will not activate donor NK cells,as they lack a sufficient cell-surface density of activating ligands for the donor NK cells.c.An alternative approach is to boost endogenous NK cell activity by treating patients with monoclonal antibodies specific for NK cell-expressed inhibitory receptors.These antibodies are designed to enhance the antitumour activity of the patients own NK cells without inducing autoimmunity.Targeting natural killer cells and natural killer T cells in cancer.NATURE REVIEWS|IMMUNOLOGY VOLUME 12|APRIL 2012 Cancer therapies targeting NK 57LAKLAK细胞的制备和应用细胞的制备和应用肿瘤患者肿瘤患者 全血细胞（淋巴细胞）全血细胞（淋巴细胞）加加IL-2，培养、扩增，培养、扩增过继性输过继性输注注抗肿瘤淋巴抗肿瘤淋巴细胞细胞加加IL-2培养、扩增培养、扩增的淋巴细胞的淋巴细胞Treatment of melanoma with LAK cells and IL-2BeforeAfterExperimental demonstration of tumor-destroying activity of LAK cells plus IL-2.Spleen cells or LAK cells,in the presence or absence of recombinant IL-2,were infused into mice with pulmonary sarcoma.The animals were evaluated