肿瘤分子生物学5. 肿瘤免疫学若干进展刘永忠文档资料.ppt

Carcinogenesis and immune microenvironments,讨论要点,免疫编辑肿瘤免疫微环境形成的细胞和分子细胞死亡或衰老和肿瘤免疫反应,2,一.肿瘤免疫编辑（CANCER IMMUNOEDITING）,3,4,Robert Schreiber,in 2001,found that immunodeficient mice are more susceptible to chemically-induced,as well as spontaneous,tumors.Shankaran,V.et al.IFN and lymphocytes prevent primary tumor development and shape tumor immunogenicity.Nature,410,11071111(2001),(A)Age-and sex-matched mice were inoculated with 100 ug MCA and monitored for tumor development for 160 days.(B)Mice housed in a specific pathogen-free facility were monitored for spontaneous tumor development between 1324 months.,5,6,7,8,Evidence for Hematopoietic immune system as anti-carcinogenic forceMOUSE MODELSDeletion of immune cells favors tumor growth,ablation of cytokine IFN,STAT1,etc,blockage of TGF,preventional or therapeutic Vaccination work in certain cancer models.,CANCER IMMUNOSURVEILLANCE IN HUMANS,Immunosuppressed transplant recipients display higher incidences of cancers.Cancer patients can develop spontaneous adaptive and innate immune responses to the tumors.(C)Infiltrating lymphocytes correlate with the favorable clinical prognosis(ovarian,colon).(d)Adoptive transfer of tumor specific T cells suppresses tumor growth.,9,Immunosuppressed transplant recipients have higher incidences of cancersBirkeland SA(1995.Int.J.Cancer 60:18389)5692 renal transplant patients from 19641982 howed increased standardized cancer incidence ratios for colon,lung,bladder,kidney,ureter,and endocrine tumors compared to the general population.Pham SM,1995.Ann.Thorac.Surg.60:162326608 cardiac transplant patients at the University of Pittsburgh between 1980 and 1993,the prevalence of lung tumors was 25-fold higher than in the general population.,Immunosuppressive intervention predisposed the transplant patients either to de novo tumor formation or allowed the outgrowth of occult tumors.,10,Donor-derived cancer,H.Myron Kauffman et al.Transplant tumor registry:Donor related malignancies.Transplantation74,358(2002).R.M.MacKie,R.Reid,B.Junor.Fatal melanoma transferred in a donated kidney 16 years after melanoma surgery.N.Engl.J.Med.348,567(2003).,Cancer patients can develop spontaneous adaptive and innate immune responses to the tumorsparaneoplastic neurologic disorders(PNDs)A human cancer immunome database(over 1000 human tumor antigens)(https:/www2.licr.org/CancerImmunomeDB/),12,13,Four classes:Differentiation Antigens,e.g.,melanocyte differentiation antigens,Mutational Antigens,e.g.,abnormal forms of p53;Overexpressed/Amplified Antigens,e.g.,HER-2/neu;Viral Antigens,e.g.,EBV and HPV;Cancer-Testis(CT)Antigens,MAGE,BAGE,GAGE;NY-ESO-1,Cancer immunosurveillance controversy A functional cancer immunosurveillance exists,however,immunocompetent individuals still develop cancer.Immunity plays a dual role in the complex Interactions between tumors and the host.Both the innate and adaptive immune systems serve not only to protect the host from tumor development but also to sculpt,or edit,the immunogenicity of tumors.,14,REFINING CANCER“IMMUNOSURVEILLANCE”TO“IMMUNOEDITING”,“Three Es of cancer immunoediting:”,Annu.Rev.Immunol.2004.22:32960,15,16,Cancer Immunoediting,Robert D.Schreiber,Science 331,1565(2011);,Immunodeficient RAG-2-/-hosts were injected with a dose of 105 tumor cells derived from wild-type 129/SvEv mice(A)or RAG-2-/-mice(B).Tumor growth is plotted as mean tumor diameter of 35 mice inoculated with each tumor.Groups of 58 immunocompetent 129/SvEv RAG-2-/-F1 mice were injected on day 0 with doses of 106 tumor cells derived from 17 individual 129/SvEv mice(C)or 20 individual RAG-2-/-mice(D)and tumor growthwas monitored as above.In(D),the dashed linesdenote tumors that grew progressively,whereas solid lines represent tumors that were rejected.,Increased immunogenicity of tumors derived from MCA-treated RAG-2-/-mice.,17,The first evidence experimentally confirms that an equilibrium process of transformed cells exists during tumor development.,18,Experimental designing,60%(9/15),The adaptive immune system promotes an equilibrium state inprimary MCA-induced sarcomas,19,Demonstration of occult cancer in immunocompetent mice in the equilibrium phase of cancer immunoediting.,20,21,Nature.2012 Feb 8;482(7385):405-9.doi:10.1038/nature10803.Expression of tumour-specific antigens underlies cancer immunoediting.DuPage M,Mazumdar C,Schmidt LM,Cheung AF,Jacks T.,Nature.2012 Feb 8;482(7385):400-4.doi:10.1038/nature10755.Cancer exome analysis reveals a T-cell-dependent mechanism of cancer immunoediting.Matsushita H,.Schreiber RD.,22,参与肿瘤免疫微环境形成的细胞 和分子,REGULATORY IMMUNE CELLS,The mammalian immune system:protecting the host from a broadrange of pathogenic microorganisms while avoiding excessive immune reactions.Understanding the mechanisms of immunological self-tolerance will provide insights into how weak immune responses,such as thoseagainst tumor antigens in cancer patients or against microbialantigens in chronic infection,can be augmented,or conversely,how strong immune responses such as graft rejection can beRestrain.,23,Two types of mechanisms,“recessive”and“dominant,”for achieving immune-tolerance and immune homeostasis.Recessive mechanism:in a cell-intrinsic manner,lymphocytes are programmed to die by apoptosis when exposed to self-antigen at an immature stage of their development in the central generative organs(the thymus for T cells and the bone marrow for B cells)Anergy in periphery,activation induced cell death Dominant mechanism:cell-extrinsic mechanism,in periphery.certain immune cells actively check the activation and expansion of aberrant or overreactive lymphocytes.,24,I.CD4+Foxp3+Regulatory T Cell,Treg,Foxp3 Member of Forkhead/winged-helix family,transcriptional factor,located on the X chromosome,Essential in immune tolerance Negatively regulation of T,B lymphocytes and NK cells,Macrophages,etc,调节性T细胞（Treg）的缺陷或过度存在与多种疾病的发生、发展相关，如自身免疫病，肿瘤，感染性疾病等。,25,Treg cell deficiency causes diseases in mice,26,IPEX：immune dysregulation,polyendocrinopathy,enteropathy,X-linked syndrome).免疫功能失调,多内分泌病变以及肠病综合征,27,Differentiation of Naive CD4+T Cells into Tregs or effector T Cells,CD25,cytotoxic Tlymphocyte antigen 4CTLA4),glucocorticoid-induced tumornecrosis factor(TNF)receptor(GITR)and OX40,were increased.,28,The tumor perturbs TReg-cell homeostasis,Nature Medicine 10,942-949(2004)Specific recruitment of regulatory T cells in ovarian carcinoma fosters immune privilege and predicts reduced survivalZou provided the first clinical evidence that Foxp3 Treg cells are crutial for the subversion of immunity to suppression in ovarian cancer patients.,29,FOXP3+Treg cells in ovarian tumor mass.,30,Regulatory T cells in the network of immunosuppression,Indoleamine 2,3-dioxygenase degrades人吲哚胺2,3-双加氧酶(IDO),31,Modulating the action of Treg in cancer therapy CHEMOTHERAPYCyclophosphamide(Metronomic low dose-regimen):T-regulatory cell,glucocorticoid-induced tumour necrosis factor receptor(GITR)and FOXP3,reversibility takes place 10 days after cyclophosphamide administration.Cancer Immunol Immunother.2007 May;56(5):641-8.Clin Exp Immunol.2007 December;150(3):523530.Fludarabine a preferential decline in CD4+CD25+Tregs was noted in patients with CLL were treated with fludarabine Blood.2005 Sep 15;106(6):2018-25.Gemcitabine In a phase I study of colon cancer patients,rise in CTLs with a concomitant decrease in Treg cells in clinical responders.,2.DIRECT TARGETING OF TREG Targeting CD25 LMB-2 a fusion protein consisting of a single-chain Fv fragment of a CD25-specific monoclonal antibody attached to a 38 kDa fragment of Pseudomonas exotoxin A.Denileukin diftitox a fusion protein combining human IL-2 and an enzymatically active domain of diphtheria toxin.Targeting CTLA-4 Two humanised anti-CTLA-4-blocking antibodies,MDX-010(Ipilimumab)and CP-675 206(Tremelimumab),have been used in phase I and II trials.,3.OTHERSTyrosine kinase inhibitors Glivec A significant dose-dependent decrease in CD69,GITR,CTLA-4 and FOXP3 expression,and IL-10 and TGF-b secretion by Tregs in patients with chronic myeloid leukemia Sunitinib(effective in the treatment of RCC)attenuates Treg numbers in miceCOX-2 inhibitor A significant increase in CD8T tumour-infiltrating T-cells and decreased expression of FOXP3 and IL-10 in the patients with colon cancer suject to an oral NSAID(indomethacin or celebrex)Aromatase inhibitor Immunomodulation of FOXP3+regulatory T cells by the aromatase inhibitor letrozole in breast cancer patients.Clin Cancer Res.2009 Feb 1;15(3):1046-51,The Regulatory T CellAssociated Transcription Factor FoxP3 Is Expressed by Tumor Cells.Cancer Research 68,3001-3009,2008,FOXP3 is an X-linked breast cancer suppressor gene and an important repressor of the HER-2/ErbB2 oncogene.Cell 2007;129:127586.,Female mice heterozygous for the scurfin mutation of the Foxp3 gene(Foxp3(sf/+)developed cancer at a high rate.The majority of the cancers with the inactivation of wild-type Foxp3 allele showed overexpression of HER-2/ErbB2.Foxp3 bound and repressed the HER-2/ErbB2 promoter.,Foxp3 expression in pancreatic carcinoma cells as a novel mechanism of immune evasion in cancer.Cancer Res 2007;67:834450,Foxp3,only functions in Treg cell lineage?,35,36,Can Treg cells do anything good for preventing cancer development or implication in cancer therapy?,Clinical Applications of Human Treg Cells in Allo-HSCT Recipients with hematological malignancies,Immunity 2009;30(5):656-665,38,Nat Med.2014 Feb 16,FOXA1+CD4Treg(PD-L1 high),II.Tumor-associated Macrophages(TAMs)A Paradoxical role of macrophages:Part of innate immunity and promoter for adaptive immune response;Macrophages are educated by the tumor microenvironment,so that they enhance tumor progression and immunosuppression.,Macrophage in colon cancer,More than 80%of clinical reports provide the evidence of a significant correlation between TAM density and poor prognosis.,39,A correlation between an abundance of TAMs and poor prognosisIn normal tissues,pathogenic challenge or wounding Expression of CSF-1;GM-CSF,and TGF-1 and chemokines CCL2,CCL7,CCL8,CCL3,CCL4 and MIF.Recruiting circulating monocytes and stimulate them to differentiate into macrophages.Macrophages,in turn,mediate immune responses,kill pathogens,stimulate angiogenesis and effect tissue repair.,40,The same things also happen for recruitment and differentiation of TAM in the context of tumorigenesis CCL2 over-expression a subsequent increase in TAMs numbers,resulted in increased tumor growth in vivo.CSF-1 deletionTAMs depletion in spontaneous polyoma virus middle T oncoprotein(PyMT)-induced mammary carcinoma.The delayed tumor angiogenesis and progression,and reduced pulmonary metastasis.,41,Pro-tumorigenic functions of tumor-associated macrophages,42,Nomenclature used previously to define MDSC:immature myeloid cells or myeloid suppressor cellsHans Schreibers group:Pekarek,L.A.,Starr,B.A.,Toledano,A.Y.,and Schreiber,H.1995.Inhibition of tumor growth by elimination of granulocytes.J.Exp.Med.181:435440.Questions arise after this surprising finding:The Gr-1specific antibody could bind and eliminate other cells in the blood.Gr-1+cells in tumor-bearing hosts were,in fact,mostly CD11b+and comprised both polymorphonuclear and mononuclear cells,including cells at different stages of maturation along the myelomonocytic differentiation pathway.,III.Myeloid-derived suppressor cells,43,Definition of MDSC:A population of myelomonocytic cells normally lacking the markers of mature myeloid cells and commonly expressing both Gr-1 and CD11b,or CD11b in mice,with a high potential to suppress immune responses in vitro and in vivo.,Li H,The Journal of Immunology,2009,182:240249.,Spleen,44,45,In vitro effects:MDSCs inhibit T cell activation(CD8+T cells more than CD4+T cells)induced by either antigens or polyclonal stimuli through an MHC-independent mechanism requiring cell-cell contact.MDSCs can take up and cross-present tumor-associated antigens in the context of MHC class I molecules in vivo,indicating that MHC-dependent responses might be relevant in vivo.,MDSC-mediated immune suppression,46,Inhibitory effects of MDSC l-Arg metabolism,47,IL-23/IL-12,IL-12 and IL-23:a small family of pro-inflammatory heterodimeric cytokines.,IL-12 promotes classical interferon-gamma producing Th1 cells and enhances cytotoxic,antimicrobial and antitumor responses.,IL-23 drives the generation of IL-17-producing CD4 T cells.,48,Overexpression of IL-23 but not of IL-12 in human cancer,TAM(CD11b+)and dendritic cells(CD11c+)isolated from mouse syngeneic tumor models expressed preferentially high amounts of IL-23p19mRNA rather than IL-12p35,49,IL-23p19 mRNA,Mice:IL-12 signaling-deficient:Il12p35-/-,Il12p40-/-,Il12R-beta1-/-and IL12R-beta2-/-IL-23 signaling-deficient:Il23p19-/-,Il12p40-/-,Il23R-/-,Il12R-beta1-/-The two-step skin carcinogenesis mouse model:-treatment with 9,10-dimethyl-1,2-benzanthracene(DMBA)once at the age of 23 months,then treated twice-weekly with the tumor promoter 12-O-tetradecanoyl-phorbol acetate(TPA)for up to 1 year.,50,Tumor resistance with reduced inflammation in IL-23-deficientanimals,51,52,IL-12 stimulates,but IL-23 decreases,immune surveillance byCD8 T cells,53,IL-23 promotes inflammatory responses such as upregulation of the matrix metalloprotease MMP9,and increases angiogenesis but reduces CD8 T-cell infiltration.IL-23 is an important molecular link between tumor-promoting pro-inflammatory processes and the failure of the adaptive immune surveillance to infiltrate tumors.,54,55,IL-17Interleukin-17A(IL-17A),CTLA-8,cloned from a murine cytotoxic T-lymphocyte hybridoma cDNA library.Murine IL-17A:63%amino acid homology with human IL-17A.Five additional related cytokines were identified(IL-17B,IL-17C,IL-17D,IL-17E also called IL-25,and IL-17F)with 1650%amino acid identity with IL-17A.,56,Th cell subsets,57,The role of IL-17A in cancer development,I.Antitumor,B16 melanoma/IL-17A KO mice model,58,59,B16 melanoma and MB49 bladder Carcinoma,IL-17 KO Enhancement of tumor growth by IL-17 involves direct effects on tumor cells and tumor-associated stromal cells.IL-17 induces IL-6 production,which in turn activates oncogenic Stat3,up-regulating prosurvival and proangiogenic genes.,IL-6 has dual functions,acting as both a pro-inflammatory and anti-inflammatory cytokine.It is secreted by T cells and macrophages to stimulate immune,especially in traumaInterleukin-6(IL-6)is responsible for the hepatic response to infections or systemic inflammation,often termed the“acute phase response.”Concentrations of IL-6 in serum are increased in situations of chronic liver inflammation including alcoholic hepatitis,HBV and HCV infections,and in patients with HCC.,Interleukin-6,60,Gender disparity in hepatocellular carcinoma(HCC),males are about three to five times more likely to develop HCC than women.A similar or even more pronounced gender disparity is observed in rodent HCC models.,61,Chemical carcinogen diethylnitrosamine(DEN)induced HCC model i