2013研究生讲座肿瘤微环境简介ppt课件

肿瘤微环境简介,苏州大学免疫学系 居颂光,？,Sid P. Kerkar and Nicholas P. Restifo. Cancer Res 2012;72:3125-3130.,肿瘤微环境,如果仅仅关注肿瘤细胞，那我们在盲人摸象,肿瘤研究肿瘤细胞研究,一个世纪的观念：肿瘤是局部组织/器官细胞异常生长形成的疾病 肿瘤是个系统性疾病 肿瘤是一种组织（间质、血管、微环境），不是一堆肿瘤细胞 免疫功能失控，肿瘤患者的免疫系统不是简单的低下： 保护肿瘤生长的免疫细胞：功能增强 抑制肿瘤细胞的免疫细胞：功能低下,肿瘤的免疫编辑,免疫成分在肿瘤发生过程中的作用,Dunn G,et al.Nature immunology. 2002.3(11):991,Elimination corresponds to immunosurveilliance,The host immune system and any tumor variant that has survived in the elimination process enter into the dynamic equilibrium: *lymphocytes and IFN- exert potent selection pressure. *A tumor bed is containing many genetically unstable and rapidly mutating tumor cells.,Survival tumor variants that have acquired insensitivity to immunologic detection and/or elimination begin to expand in uncontrol manner.,Dunn G,et al.Nature immunology. 2002.3(11):991,Solid tumors reach a certain size: Grow invasively, require an enhanced blood supply. Inflammatory signals leading to recruitment of cells of innate immune system: macrophages, DCs, -T, NK and NKT. Produce IFN-,The initial IFN- starts a cascade of immune response: 1.Induction of chemokines:CXCL10,CXCL9,CXCL11.Block neovascularization of tumor, recruit macrophages, DCs ,-T, NK and other immune cells. 2.Antiproliferation of developing tumor. 3.The activation of cytocidal activity in macrophages and NK cells entering the tumor. Dead tumor cells or tumor cells debris are ingested by DC and are trafficked to the draining lymph node.,1.Tumor growth is kept in check by the cytocidal NK and mocrophages. 2.CD4+T and CD8+T cells that are specific for tumor antigens are developing in draining lymph node.,Tumor specific CD4+T and CD8+T cells home to tumor site along a chemokine gradient .,Dunn G,et al.Nature immunology. 2002.3(11):991,Elimination corresponds to immunosurveilliance,The host immune system and any tumor variant that has survived in the elimination process enter into the dynamic equilibrium: *lymphocytes and IFN- exert potent selection pressure. *A tumor bed is containing many genetically unstable and rapidly mutating tumor cells.,Survival tumor variants that have acquired insensitivity to immunologic detection and/or elimination begin to expand in uncontrol manner.,肿瘤细胞,肿瘤抗原,Olivera J. Finn, Ph.D.,N Engl J Med 2008;358:2704-15,Olivera J. Finn, Ph.D.,N Engl J Med 2008;358:2704-15,肿瘤抗原的加工提呈,Olivera J. Finn, Ph.D.,N Engl J Med 2008;358:2704-15,肿瘤干细胞,Knutson K, et al. Cancer Immunol Immunother 2005, 54: 721728,APC TH Treg Cytoxic T cells Others,IFN IL-2 IL-12 Granzyme Perforin Abs others,Tumor microenvironment:,N Engl J Med 2006;355:1253 - 1261.,Cancer stem cell,Nature Reviews Cancer doi:10.1038/nrc1232,Tumor Heterogeneity and Cancer Stem Cells,肿瘤微环境中的免疫稳态失衡,Nature.2005,5:263,肿瘤患者的免疫状态是有别于正常人的免疫稳态的。,肿瘤微环境中的免疫细胞,TH17 cells in tumour immunity and Immunotherapy. NATuRE REVIEWS | Immunology. doi:10.1038/nri2742,Differentiation of helper T cell subsets,TH17 cells and antitumour immunity,肿瘤微环境中的抑制性成分Treg细胞,Treg细胞定义,1995由Sakaguchi等发现了机体重要的负性免疫调节成分调节性CD4+CD25+ T细胞（regulatory T cell, Treg）。随后的研究根据细胞内标记分子Foxp3和细胞膜分子CD127表达与否，将经典的Treg细胞定义为CD4+CD25+ Foxp3+CD127low/neg T细胞。 通过细胞间接触依赖机制发挥作用或分泌IL-10和TGF-等细胞因子，Treg细胞抑制CD4+25-T细胞、CD8+T细胞和树突状细胞等其他免疫细胞的活性和功能。,大量的研究报道表明:CD4+CD25+Treg细胞是抑制机体抗肿瘤免疫应答的关键成分之一，例如： Woo等发现在非小细胞肺癌和卵巢癌患者的肿瘤浸润淋巴细胞中Treg的含量明显增多； 在胃肠道肿瘤患者中CD4+CD25-T细胞明显减少而Treg细胞数量增多，而且Treg细胞数量的增高与较差的预后和较低的生存率高度相关； Curiel 等报道，在卵巢癌中Treg细胞能抑制T细胞介导的特异性抗肿瘤免疫应答，并且有利于肿瘤的生长； 因此研究者希望通过减少Treg细胞的数量或降低其活性来开辟治疗肿瘤免疫治疗的新途径。例如，以Foxp3为靶点清除荷瘤小鼠体内的Treg细胞后肿瘤会消退。,Treg细胞的作用机制,非小细胞性肺癌（NSCLC）与Treg新亚群,Fig 1,Fig.2,Fig 3,Fig 4,CD8+T细胞在肿瘤免疫中的作用,肿瘤微环境中的“正性”免疫成分,Gadd45-/- mice or Het/WT mice were injected into intradermally of flank skin with 2.5X104 B16 cells/mouse.,Tumor sizes were measured every 2 days.,Tumor inject site,Gadd45b KO mice were challenged with melanoma cells: B16,Freeze down the tissue of tumor site.,Fig 4 Gadd45-/- mice or WT mice were injected subcutaneously with 2.5X104 B16 cells/mouse, tumor sizes were measured every 2 days. These data are from 3 separated experiments.,5.Both WT and Gadd45b KO T cells are able to infiltrate into tumor sites.,Gadd45-/- mice or WT mice were injected intradermally of flank skin with 5X104 B16 cells/site/20ul PBS,Skin of injected sites or opposite sites was resected and freeze down immediately,Spleen Cells Adoptive Transfer and B16 inoculation,Tumor inject site,Check tumor formation from injected sites,500 Rad,WT,KO,500 Rad,Donor,Host,6. Gadd45 is important for lymphocytes in tumor surveillance,P0.05,*,CD4+T Cells Adoptive Transfer and B16 inoculation,500 Rad,WT CD4+T,KO CD4+T,500 Rad,Donor,Host,WT splenocytes depleted CD4+T,WT splenocytes deplete CD4+T,+,+,7. Gadd45b KO does not affect function of CD4+T cells in tumor surveillance,KO: Transfer with CD4+T cell depleted spleen cells from WT mice + CD4+T cells from Gadd45b KO mice,PBS: Only transfer PBS,WT: Transfer with CD4+T cell depleted spleen cells from WT mice + CD4+T cells from Gadd45b Het or WT mice,CD8+T Cells Adoptive Transfer and B16 inoculation,500 Rad,WT CD8+T,KO CD8+T,500 Rad,Donor,Host,WT splenocytes depleted CD8+T,WT splenocytes deplete CD8+T,+,+,8. Gadd45b KO causes function defect of CD8+T cells in tumor surveillance,KO: Transfer with CD8+T cell depleted spleen cells from WT mice + CD8+T cells from Gadd45b KO mice,PBS: Only transfer PBS,WT: Transfer with CD8+T cell depleted spleen cells from WT mice + CD8+T cells from Gadd45b Het or WT mice,P0.05,*,Gadd45b is expressed in CD8+T cells during their activation,Figure 1 Gadd45b message is induced by TCR signaling and costimulation. Naive CD8+ T cells were stimulated with platebound anti-CD3 (-CD3) plus -CD28 and Th1 condition for 0h,1 h, 12h, 24h, 48h and 96h. The Gadd45b message was detected with RT-PCR.,2. Gadd45 is important for the function of CD8+T cells.,IL12/IL18,-CD3,MAP kinase profile activated in CD8+T cells,-CD3 (10 g/ml),IL-12(10 ng/ml) + IL-18 (30 ng/ml),Figure 3 Gadd45 mediates p38 activation by TCR signaling or IL-12 plus IL-18 in CD8+T cells. (a) Effector CD8+T cells were stimulated (times, above lanes) with plate-bound anti-CD3 (-CD3; 10 g/ml) or (b) IL-12(10 ng/ml) and IL-18 (30 ng/ml). Cell extracts were analyzed by immunoblot with anti-phospho-ERK (-phospho-ERK), -phospho-JNK and -phospho-p38. p-, phosphorylated.,Gadd45b KO causes function defect of tumor infiltrating CD8+T cells,Fig 5 Gadd45 KO cause tumor infiltrating CD8+T cell function defect in vivo. Tumor mass was removed at day 13 after B16 F0 cells inoculation and digested with collagenase I. Cells were stimulated with phorbol 12-myristate 13-acetate (50 ng/ml) and ionomycin (500 ng/ml) for 6 h and incubated for the last 3 h with Brefeldin A. Cells producing IFN- was identified with intracellular cytokine staining.,Type, Density, and Location of Immune Cells Within Human Colorectal Tumors Predict Clinical Outcome,Galon J, et al. 29 SEPTEMBER 2006 VOL 313 SCIENCE,Type, Density, and Location of Immune Cells Within Human Colorectal Tumors Predict Clinical Outcome,Galon J, et al. 29 SEPTEMBER 2006 VOL 313 SCIENCE,(B) Comparison of the mean of immune cell densities in the CT and IM from patients with tumor recurrence (black bars) or without tumor recurrence (white bars).,肿瘤微环境中的免疫分子,肿瘤微环境中的Immune checkpoints,NATURE REVIEWS | CANCER 2012 doi:10.1038/nrc3239,Multiple co-stimulatory and inhibitory interactions regulate T cell responses,Immune checkpoints regulate different components in the evolution of an immune response,Two general mechanisms of expression of immune-che