DYNAMICS AND TUNING OF THE MHC II PRESENTED PEPTIDOME

MHC II 呈递肽段的动力学和调节

• 批准号: 10468682
• 负责人: LAURA SANTAMBROGIO
• 金额: $ 79.55万
• 依托单位: WEILL MEDICAL COLL OF CORNELL UNIV
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-09-19 至 2024-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10468682
• 关键词: Address; Adolescent; Affect; Affinity; Anatomy; Antigen Presentation; Antigen-Presenting Cells; Attention; Autoantigens; Autoimmune Diseases; Autoimmunity; Binding; Binding Proteins; Biological Assay; CD4 Positive T Lymphocytes; Cell Culture Techniques; Cell Maturation; Cervical; Chromogranins; Chronic Childhood Arthritis; Clonal Expansion; Collagen; Collagen Type II; Crohn' s disease; Dendritic Cells; Development; Diabetes Mellitus; Disease; Epitopes; Equilibrium; Generations; Goals; Harvest; Histones; Immune; Immune system; Immunity; Immunologics; In Vitro; Inbred NOD Mice; Individual; Infection; Inflammation; Inflammatory; Insulin; Insulin-Dependent Diabetes Mellitus; Isotope Labeling; Label; Location; Maintenance; Malignant Neoplasms; Mass Spectrum Analysis; Measurement; Mediating; Methodology; Modeling; Monitor; Multiple Sclerosis; Mus; Organ; Pancreas; Pathologic; Pathway interactions; Peptides; Phase; Physiological; Population; Post-Translational Protein Processing; Prealbumin; Prediabetes syndrome; Process; Proteins; Proteomics; Reproducibility; Rest; Rheumatoid Arthritis; Salmonella typhimurium; Sampling; Serum Proteins; Source; Stimulus; Surveys; System; Systemic Lupus Erythematosus; T cell response; T-Lymphocyte; Testing; Time; Tissues; Tuberculosis; adaptive immune response; antigen processing; autoreactive T cell; base; central tolerance; draining lymph node; experimental study; extracellular; in vivo; in vivo monitoring; instrumentation; insulitis; islet; lymph nodes; mesenteric lymph node; pathogen; peripheral tolerance; prevent; proteomic signature

This application addresses fundamental open questions on MHC-II peptidome selection and composition, namely, the plasticity of the MHC-II peptidome displayed by conventional dendritic cells (cDC) at different organ locations and under physiological and pathological conditions. Overall our analysis will generate a quantitative mapping of (i) how the MHC-II peptidome reflects the “organ- specific proteomic signature” from where the cDC originates and (ii) the relationship between presented MHC II peptidome in resting and inflammatory conditions, for both self and non-self-epitopes. Aim 1 is to understand how qualitative and quantitative changes in MHC II epitope copy number presented in local lymph node contribute to the balance between tolerance and autoimmunity. The MHC II peptidome will be compared among cDC harvested from lymph nodes draining different organs and quantitative mass spectrometry will be utilized to monitor changes in the immune-peptidome and peptide abundance across tissue cDC. In particular we will quantify the peptide copy numbers on local cDC for tissue specific self antigens, known to be targets in autoimmune diseases and whose auto-reactive T cells are found in the periphery, including MBP, PLP, MOG, (multiple sclerosis) D1-antitrypsin (Crohn's disease), Insulin, GAD65, Chromogranin, (diabetes type I), histones, (systemic lupus erythematosus) and collagen II (rheumatoid arthritis) and Transthyretin (RA and juvenile idiopathic arthritis). Additionally we will use the well- characterized NOD mice Type 1 Diabetes model to answer the question of whether when mice cross the threshold between physiological conditions and pre-diabetic insulitis quantitative and qualitative changes in the I-Ag7-peptidome associated with disease development (insulin, chromogranin, GAD65, S100), are observed. Aim 2 is to understand the consequences of infection and inflammation on the MHC II-self peptidome . Using quantitative isotope labeling and label-free proteomic approaches we will monitor the in vivo presented self- and noself-peptidomes presented cDC in resting mice and after infection with Mycobaterium tuberculosis or Salmonella typhimurium, to determine how the cDC maturation process changes the presented MHC-II self peptidome. We will determine any changes in MHC II affinity and HLA-DM sensitivity in the MHC II self- peptidome induced by infection, and differences in the MHC II antigen processing pathways in immature and pathogen-matured cDC. Finally, we evaluate how changes in post-translational modification processing induced during DC maturation impact the presented peptidome. Overall these studies will determine how MHC II epitope copy number presented in local lymph node can contributes to the balance between tolerance and autoimmunity and what are the consequences of infection and inflammation on the displayed MHC II self peptidome.

该申请介绍有关MHC-II Pepperome选择和组成的基本开放问题， 也就是说，常规树突状细胞（CDC）在不同 器官位置以及生理和病理条件下。 总体而言，我们的分析将产生（i）MHC-II Pepperome如何反映“器官”的定量映射。 特定的蛋白质组学签名”从疾病预防控制中心起源的地方和（ii）提出的MHC II之间的关系 肽组在静止和炎症条件下，用于自我和非自我症状。 目标1是了解MHC II表位副本编号的定性和定量变化如何 在局部淋巴结中提出的耐受性和自身免疫性之间的平衡有助于。 MHC II Pepperidome将在从排出不同器官和的淋巴结中的CDC中进行比较 定量质谱法将用于监测免疫肽组和胡椒的变化 跨组织CDC的抽象。特别是我们将在局部CDC上量化组织的肽拷贝数 特定的自抗原，已知是自身免疫性疾病的靶标，并且发现其自身反应性T细胞 在外围，包括MBP，PLP，MOG，（多发性硬化症）D1-抗胰蛋白酶（克罗恩病），胰岛素， GAD65，促铬素（糖尿病I型），组蛋白，（全身性红斑狼疮）和胶原蛋白II （类风湿关节炎）和甲状腺素蛋白（RA和少年特发性关节炎）。此外，我们将使用 表征了1型糖尿病模型的点头小鼠，以回答何时越过小鼠的问题 身体状况与糖尿病前胰岛炎之间的阈值定量和定性变化 观察到与疾病发育相关的I-AG7肽组（胰岛素，铬烷蛋白，GAD65，S100）。 目的2是了解感染和感染对MHC II并发性胡椒的后果。 使用定量同位素标记和无标记的蛋白质组学方法，我们将监测呈现的体内 自我和NOSELF肽组在静止小鼠中呈现CDC，并在感染结核病后感染了CDC 或沙门氏菌伤寒，确定CDC的成熟过程如何改变了呈现的MHC-II自我 肽组。我们将确定MHC II自我自我的MHC II亲和力和HLA-DM敏感性的任何变化 通过感染诱导的肽组，以及未成熟和 病原体成熟CDC。最后，我们评估翻译后修改处理的变化 在直流成熟期间诱导的影响肽组。 总的来说，这些研究将确定在局部淋巴结中显示的MHC II发作拷贝数如何 有助于耐受性和自身免疫之间的平衡以及感染的后果是什么 和显示的MHC II自肽组上的炎症。