Role of autophagy-related protein Vps34 in antigen presentation and self-tolerance

自噬相关蛋白 Vps34 在抗原呈递和自我耐受中的作用

• 批准号: 10214468
• 负责人: Luc Van Kaer
• 金额: $ 52.54万
• 依托单位: VANDERBILT UNIVERSITY MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-08-15 至 2024-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10214468
• 关键词: 1-Phosphatidylinositol 3-Kinase; Abbreviations; Allergens; Animals; Antigen Presentation; Antigen Presentation Pathway; Antigens; Apoptotic; Autoantigens; Autoimmunity; Autophagocytosis; Biological Assay; CD8-Positive T-Lymphocytes; Cell Culture Techniques; Cell physiology; Cells; Complement; Cross Presentation; Defect; Dendritic Cells; Development; Diabetes Mellitus; Disease; Eating; Endocytosis; Exhibits; FOXP3 gene; Future; Generations; Genes; Genetic Engineering; Goals; Histocompatibility Antigens; Histocompatibility Antigens Class II; Human; Hypersensitivity; Immune; Immune response; Immune system; Immunoglobulins; Immunotherapy; In Vitro; Infectious Agent; Islets of Langerhans; Life; Listeria monocytogenes; Lysosomes; MHC Class I Genes; MHC Class II Genes; Malignant Neoplasms; Mediating; Mediator of activation protein; Methods; Modeling; Molecular; Mucins; Mus; Oral; Pathway interactions; Peripheral; Phagocytosis; Pharmacology; Phenotype; Phosphatidylserines; Play; Population; Process; Property; Proteins; Proteolysis; Publishing; Quality Control; Regulatory T-Lymphocyte; Reporter; Reporter Genes; Role; Self Tolerance; Signal Transduction; T cell response; T-Cell Development; T-Cell Receptor; T-Lymphocyte; Testing; Thymic epithelial cell; Thymus Gland; Tissues; Transgenic Mice; Vaccines; Vacuolar Protein Sorting; Vesicle; Work; adaptive immune response; aged; anti-dsDNA antibodies; antigen-specific T cells; base; cell type; cytokine; human disease; improved; in vivo; neoantigens; neoplastic cell; novel vaccines; organ transplant rejection; pathogenic bacteria; pathogenic microbe; peripheral tolerance; prevent; receptor; response; trafficking; tumor; uptake; vaccine development

PROJECT SUMMARY The presentation of self and foreign antigens to T lymphocytes involves a variety of cellular processes, including proteolysis, endocytosis, phagocytosis, vesicle trafficking and autophagy. Our recent studies have investigated the role of autophagy, a self-degradation mechanism, and related cellular processes in MHC class I- and class II-restricted antigen presentation to T lymphocytes, with the long-term goal of manipulating these pathways to prevent or treat human disease. We have focused on a key player in autophagy, the class III phosphatidylinositol- 3 kinase (PI3K) vacuolar protein sorting 34 (Vps34). Our published and unpublished preliminary studies have provided evidence for a key role of Vps34 in both MHC class I- and class II-restricted antigen presentation by dendritic cells (DCs), which are critical players in the induction of tolerance against self-antigens and for initiating adaptive immune responses against foreign antigens. We have shown that the classical MHC class I and class II antigen presentation pathways are enhanced in Vps34-deficient DCs, and that these cells harbor a specific defect in the cross-presentation of apoptotic cell-associated antigens to MHC class I-restricted CD8+ T cells. Moreover, using mice carrying a selective deficiency of Vps34 in their thymic epithelial cells (TECs), we have shown a critical role of Vps34-mediated cellular processes in intrathymic T lymphocyte development. Guided by this scientific premise we propose the overall hypothesis that Vps34-mediated, autophagy-related processes play critical roles in presenting antigens to T lymphocytes and promoting self-tolerance. We will test this hypothesis in the following specific aims: Aim 1 will investigate the contribution of Vps34-mediated functions in unconventional MHC class I- and class II-restricted antigen presentation, and will explore the autophagy-dependent and -independent molecular mechanisms involved. Aim 2 will interrogate the role of Vps34 in the intrathymic development and selection of the T cell repertoire. Aim 3 will explore the role of Vps34 in maintaining peripheral tolerance and controlling T lymphocyte responses against autoantigens. In these studies, we will employ a variety of experimental approaches, including in vitro and in vivo antigen presentation assays, and various genetically engineered animals. As global Vps34 gene-deficiency is incompatible with life, we will employ mice selectively deficient in Vps34 expression in either DCs or TECs. We will complement these studies with cultures of murine and human cells using pharmacological modulators of Vps34 function. We will also take advantage of a variety of reporter animals and transgenic mice expressing neo self-antigens and/or antigen- specific T cell receptors. Collectively, these proposed studies will provide us with a comprehensive mechanistic view of the contribution of the autophagy-related protein Vps34 in antigen presentation, T cell development and selection, peripheral tolerance and T cell immune responsiveness. These proposed studies will be instrumental for the development of improved vaccines and immunotherapies against microbial pathogens and tumors, and for devising methods to induce tolerance against self or foreign antigens.

项目总结 将自身和外源抗原呈递给T淋巴细胞涉及多种细胞过程，包括 蛋白分解、内吞、吞噬、囊泡运输和自噬。我们最近的研究调查了 自噬，一种自我降解机制，以及相关的细胞过程在MHC I类和CLASS中的作用 II-限制性抗原递呈给T淋巴细胞，长期目标是操纵这些途径来 预防或治疗人类疾病。我们关注的是自噬中的一个关键因素，即III类磷脂酰肌醇- 3激酶(PI3K)空泡蛋白分选34(Vps34)。我们已发表和未发表的初步研究有 为Vps34在MHC I类和II类限制性抗原提呈中的关键作用提供了证据 树突状细胞(DC)，在诱导对自身抗原的耐受和启动 针对外来抗原的适应性免疫反应。我们已经证明了经典的MHC I类和类 在Vps34缺陷的DC中，II抗原提呈通路被增强，并且这些细胞具有特定的 凋亡细胞相关抗原与MHC-I类限制性CD8+T细胞的交叉呈递存在缺陷。 此外，使用在胸腺上皮细胞(TECs)中携带Vps34选择性缺陷的小鼠，我们有 显示了Vps34介导的细胞过程在胸腺内T淋巴细胞发育中的关键作用。指导原则 在这一科学前提下，我们提出了Vps34介导的、与自噬相关的总体假设 过程在将抗原呈递给T淋巴细胞和促进自我耐受方面起着关键作用。我们 我将在以下具体目标中检验这一假设：目标1将调查Vps34介导的贡献 在非常规MHC I类和II类限制性抗原提呈中的作用，并将探索 涉及自噬依赖和独立的分子机制。目标2将询问Vps34的作用 在胸腺内发育和选择T细胞谱系。目标3将探索Vps34在 维持外周耐受和控制T淋巴细胞对自身抗原的反应。在这些研究中， 我们将采用各种实验方法，包括体外和体内的抗原提呈分析， 以及各种基因工程动物。由于全球Vps34基因缺失与生命不相容，我们将 在DC或TECs中使用选择性缺乏Vps34表达的小鼠。我们会补充这些研究。 使用Vps34功能的药理调节剂对小鼠和人类细胞进行培养。我们也会拿到 多种报告动物和表达neo自身抗原和/或抗原的转基因小鼠的优势- 特异性T细胞受体。总的来说，这些拟议的研究将为我们提供一个全面的机制 自噬相关蛋白Vps34在抗原提呈、T细胞发育和免疫调节中的作用 选择、外周耐受性和T细胞免疫反应性。这些拟议的研究将是有用的 用于开发针对微生物病原体和肿瘤的改进疫苗和免疫疗法，以及 设计方法以诱导对自身或外来抗原的耐受性。