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

基本信息

批准号：
10448553
负责人：
Luc Van Kaer
金额：
$ 7.17万
依托单位：
VANDERBILT UNIVERSITY MEDICAL CENTER
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-08-15 至 2023-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10448553
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

项目摘要

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.

项目概要将自身抗原和外源抗原呈递给 T 淋巴细胞涉及多种细胞过程，包括蛋白水解、内吞作用、吞噬作用、囊泡运输和自噬。我们最近的研究调查了自噬、自我降解机制以及 MHC I 类和 I 类相关细胞过程的作用 II-限制性抗原呈递至 T 淋巴细胞，长期目标是操纵这些途径预防或治疗人类疾病。我们重点关注自噬中的一个关键角色，即 III 类磷脂酰肌醇 - 3 激酶 (PI3K) 液泡蛋白分选 34 (Vps34)。我们已发表和未发表的初步研究为 Vps34 在 MHC I 类和 II 类限制性抗原呈递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性和启动免疫反应中发挥着关键作用针对外来抗原的适应性免疫反应。我们已经证明经典的 MHC I 类和 I 类 II 抗原呈递途径在 Vps34 缺陷的 DC 中得到增强，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 细胞交叉呈递的缺陷。此外，利用胸腺上皮细胞 (TEC) 选择性缺乏 Vps34 的小鼠，我们发现显示了 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。指导者在此科学前提下，我们提出了总体假设，即 Vps34 介导的、自噬相关的这些过程在向 T 淋巴细胞呈递抗原和促进自我耐受方面发挥着关键作用。我们将在以下具体目标中检验这一假设：目标 1 将研究 Vps34 介导的贡献在非常规 MHC I 类和 II 类限制性抗原呈递中发挥作用，并将探索涉及自噬依赖和独立的分子机制。目标 2 将质疑 Vps34 的作用 T 细胞库的胸腺内发育和选择。目标 3 将探讨 Vps34 在以下方面的作用：维持外周耐受性并控制 T 淋巴细胞针对自身抗原的反应。在这些研究中，我们将采用各种实验方法，包括体外和体内抗原呈递测定，以及各种基因工程动物。由于全球Vps34基因缺陷与生活不相容，我们将使用 DC 或 TEC 中选择性缺乏 Vps34 表达的小鼠。我们将补充这些研究使用 Vps34 功能的药理学调节剂培养小鼠和人类细胞。我们还将采取各种报告动物和表达新自身抗原和/或抗原的转基因小鼠的优点特异性T细胞受体。总的来说，这些拟议的研究将为我们提供全面的机制自噬相关蛋白 Vps34 在抗原呈递、T 细胞发育和选择、外周耐受性和 T 细胞免疫反应。这些拟议的研究将很有帮助开发针对微生物病原体和肿瘤的改进疫苗和免疫疗法，以及发明了诱导针对自身或外来抗原的耐受性的方法。