Regulation of Innate Dendritic Cell CTLA-4

先天树突状细胞 CTLA-4 的调节

• 批准号: 10747694
• 负责人: WILLIAM Karl DECKER
• 金额: $ 47.88万
• 依托单位: BAYLOR COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-03-13 至 2028-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10747694
• 关键词: ATAC-seq; Ablation; Adoptive Transfer; Age; Alopecia; Animals; Antigens; Autoimmune Diseases; B-Lymphocytes; Binding; Biological; C57BL/6 Mouse; CCAAT-Enhancer-Binding Proteins; CD8-Positive T-Lymphocytes; CD8B1 gene; CTLA4 gene; Cell Communication; Cell Proliferation; Cell Separation; Cell secretion; Cells; Cellular Immunity; Cellular biology; Cessation of life; ChIP-seq; Coculture Techniques; Communicable Diseases; Complex; Cues; Data; Dendritic Cells; Detection; Disease; Down-Regulation; Equilibrium; Event; Exhibits; Failure to Thrive; Formulation; Funding; GATA3 gene; Genetic Transcription; Genomics; Health; Human; ITGAX gene; Immune; Immune response; Immune system; In Vitro; Incubated; Infectious Agent; Interferon Type II; Intervention; Knock-out; LoxP-flanked allele; Lymphocytic Infiltrate; Lymphoid; Lymphoid Tissue; Malignant Neoplasms; Mass Spectrum Analysis; Mediating; Mus; Myelogenous; Myeloid Cells; Natural Immunity; Pancreas; Pathway interactions; Peripheral; Peripheral Blood Mononuclear Cell; Personal Satisfaction; Peyer' s Patches; Pharmacologic Substance; Phenotype; Play; Population; Process; Proliferating; Proteins; Publishing; Regulation; Regulatory T-Lymphocyte; Resolution; Role; Shapes; Signal Transduction; Small Interfering RNA; T-Lymphocyte; Testing; Thymus Gland; Tissues; Transcriptional Regulation; Treatment Protocols; Tumor Immunity; Up-Regulation; Vesicle; Virus Diseases; Work; antiviral immunity; arm; cell type; central tolerance; conditional knockout; druggable target; exosome; fluorescence imaging; immune activation; immune checkpoint; immune function; immunoregulation; in vivo; knock-down; knockout animal; lymphoid organ; monocyte; novel; novel vaccines; paracrine; polarized cell; programs; promoter; response; therapeutic target; thymocyte; transcriptome sequencing; transmission process; vaccination strategy

Abstract Myeloid dendritic cells (DC) are a critical lineage of innate immunity that serve as a principal point of contact and crosstalk between the innate and adaptive arms of the immune system. CTLA-4 is one of the best characterized of the immune checkpoint proteins, molecules that serve to balance, regulate, and fine-tune immune activation with homeostatic inhibition. CTLA-4 is expressed by all major lymphoid lineage effectors; however, its function- ality has been best characterized in T-cells where it exhibits both cell-extrinsic and cell-intrinsic regulatory func- tions. Until recently, very little was known about CTLA-4 expression or function in non-lymphoid cell types, par- ticularly the myeloid lineage dendritic cell subsets. In the original iteration of this renewal application, we provided preliminary data demonstrating that DC-secreted CTLA-4+ exosomes could bind B7 in paracrine fashion, leading to vesicle internalization and subsequent downregulation of B7 expression among bystander DC that internalized the CTLA-4+ exosomes. Conversely, knockdown of DC-expressed CTLA-4 resulted in a dramatic upregulation of co-cultured CD8+ cell proliferation in vitro as well as enhanced antitumor and antiviral immunity in vivo. These discoveries and concomitant characterization of myeloid CTLA-4 expression signified a paradigm shift in the understanding of CTLA-4’s role in immune governance as well as the mechanisms through which innate and adaptive crosstalk occur. Subsequent data indicated that the expression of DC CTLA-4 is modulated in response to TH polarizing cues and that regulation in DC appears to be governed in part by the transcription factors GATA3 and C/EBP-b. Further, conditional ablation of CTLA-4 in the C57BL/6 background revealed potential new roles for DC expressed CTLA-4 in regulatory processes in the thymus and in other lymphoid tissues including Peyer’s patches. These novel and exciting data have allowed formulation of a refined overarching hypothesis that DC- secreted CTLA-4+ exosomes act as effector vehicles that shape downstream TH polarization of adaptive re- sponses as dictated by DC detection of innate signaling cues. By means of three independent aims we will test this overarching hypothesis by i) defining the role of TH polarizing cues in the governance of DC CTLA-4 expres- sion and the governance of the CEBP/b and GATA3 transcriptional regulators, ii) defining the mechanisms through which DC-secreted CTLA-4+ and CTLA-4neg exosomes regulate downstream adaptive TH polarization, and iii) defining the manner by which DC CTLA-4 expression modulates central tolerance by interrogating the regulatory T-cell deficits observed in the CD11c-Cre CTLA-4flox/flox C57BL/6 mouse. Completion of these inde- pendent aims will further elucidate the novel regulatory role of myeloid CTLA-4, furthering the ability to synthesize effective and powerful vaccination strategies while characterizing critical druggable target interactions and en- hancing the understanding of complex biological pathways.

摘要 髓样树突状细胞（DC）是先天免疫的关键谱系，其充当主要接触点， 免疫系统的先天和适应性臂之间的串扰。CTLA-4是最好的表征之一 免疫检查点蛋白，分子，用于平衡，调节和微调免疫激活 体内平衡抑制CTLA-4由所有主要淋巴系效应物表达;然而，其功能- 在T细胞中，它表现出细胞外在和细胞内在的调节功能， 选择。直到最近，对CTLA-4在非淋巴细胞类型中的表达或功能知之甚少， 特别是髓系树突状细胞亚群。在这个更新应用程序的原始迭代中，我们提供了 初步数据表明，DC分泌的CTLA-4+外泌体可以旁分泌方式结合B7， 与小泡内化和随后的B7表达下调有关， CTLA-4+外泌体。相反，DC表达的CTLA-4的敲低导致DC表达的CTLA-4的显著上调。 体外共培养的CD 8+细胞增殖以及体内增强的抗肿瘤和抗病毒免疫。这些 髓系CTLA-4表达的发现和伴随的特征标志着髓系CTLA-4表达的范式转变。 理解CTLA-4在免疫管理中的作用以及先天和 发生自适应串扰。随后的数据表明，DC CTLA-4的表达是在应答中调节的。 TH极化线索，DC中的调节似乎部分由转录因子GATA 3控制。 和C/EBP-b。此外，在C57 BL/6背景中条件性消融CTLA-4揭示了潜在的新作用。 DC在胸腺和其他淋巴组织（包括Peyer’s）的调节过程中表达CTLA-4 补丁.这些新的和令人兴奋的数据已经允许制定一个完善的总体假设，DC- 分泌的CTLA-4+外泌体作为效应器载体，其形成适应性再表达的下游TH极化。 由先天信号线索的DC检测决定的自发。通过三个独立的目标，我们将测试 i）定义TH极化线索在DC CTLA-4表达调控中的作用， 锡永CEBP/B和GATA 3转录调节因子的控制，ii）定义机制 DC分泌的CTLA-4+和CTLA-4-neg外泌体通过其调节下游适应性TH极化， 和iii）通过询问DC CTLA-4表达调节中枢耐受性的方式， 在CD 11 c-Cre CTLA-4flox/flox C57 BL/6小鼠中观察到的调节性T细胞缺陷。完成这些指标- 待决目标将进一步阐明髓样CTLA-4的新的调节作用，促进合成CTLA-4的能力。 有效和强大的疫苗接种策略，同时表征关键的可药物靶向相互作用， 促进对复杂生物途径的理解。

项目成果

Cancer Immunotherapy: Historical Perspective of a Clinical Revolution and Emerging Preclinical Animal Models.

• DOI: 10.3389/fimmu.2017.00829
• 发表时间: 2017
• 期刊: Frontiers in immunology
• 影响因子: 7.3
• 作者: Decker WK;da Silva RF;Sanabria MH;Angelo LS;Guimarães F;Burt BM;Kheradmand F;Paust S
• 通讯作者: Paust S

Beyond T-Cells: Functional Characterization of CTLA-4 Expression in Immune and Non-Immune Cell Types.

• DOI: 10.3389/fimmu.2020.608024
• 发表时间: 2020
• 期刊: Frontiers in immunology
• 影响因子: 7.3
• 作者: Oyewole-Said D;Konduri V;Vazquez-Perez J;Weldon SA;Levitt JM;Decker WK
• 通讯作者: Decker WK