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: 10016167
• 负责人: LAURA SANTAMBROGIO
• 金额: $ 82.04万
• 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 2023-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10016167
• 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.

该应用解决了有关 MHC-II 肽组选择和组成的基本开放问题， 即传统树突状细胞 (cDC) 在不同条件下表现出的 MHC-II 肽组的可塑性 器官位置以及生理和病理条件下的情况。 总体而言，我们的分析将生成 (i) MHC-II 肽组如何反映“器官- 特定的蛋白质组特征”来自 CDC 的起源地和 (ii) 所呈现的 MHC II 之间的关系 静息和炎症条件下的肽组，包括自身和非自身表位。 目标 1 是了解 MHC II 表位拷贝数的定性和定量变化 存在于局部淋巴结有助于耐受性和自身免疫之间的平衡。这 MHC II 肽组将与从引流不同器官的淋巴结收集的 cDC 进行比较 定量质谱法将用于监测免疫肽组和肽的变化 跨组织 CDC 的丰度。特别是，我们将量化组织的本地 cDC 上的肽拷贝数 特定的自身抗原，已知是自身免疫性疾病的靶标，并且发现了其自身反应性 T 细胞 外周，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、嗜铬蛋白（I 型糖尿病）、组蛋白（系统性红斑狼疮）和 II 型胶原蛋白 （类风湿性关节炎）和运甲状腺素蛋白（RA 和幼年特发性关节炎）。此外，我们将使用井 NOD 小鼠 1 型糖尿病模型的特征在于回答小鼠何时跨越 1 型糖尿病模型的问题 生理状况与糖尿病前期胰岛炎之间的量和质变化之间的阈值 观察到与疾病发展相关的 I-Ag7-肽组（胰岛素、嗜铬粒蛋白、GAD65、S100）。 目标 2 是了解感染和炎症对 MHC II 自身肽组的影响。 使用定量同位素标记和无标记蛋白质组学方法，我们将监测体内呈现的 自体和非自体肽组在静息小鼠和结核分枝杆菌感染后呈现 cDC 或鼠伤寒沙门氏菌，以确定 cDC 成熟过程如何改变呈现的 MHC-II 自身 肽组。我们将确定 MHC II 自身亲和力和 HLA-DM 敏感性的任何变化。 感染诱导的肽组，以及未成熟和未成熟 MHC II 抗原加工途径的差异 病原体成熟的cDC。最后，我们评估翻译后修饰处理的变化如何 DC 成熟期间诱导的影响呈现的肽组。 总的来说，这些研究将确定局部淋巴结中呈现的 MHC II 表位拷贝数如何影响 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 以及显示的 MHC II 自肽组上的炎症。