Effects of Glycation and Carbonylation on MHC II-restricted immunity

糖化和羰基化对 MHC II 限制性免疫的影响

• 批准号: 10335198
• 负责人: LAURA SANTAMBROGIO
• 金额: $ 74万
• 依托单位: WEILL MEDICAL COLL OF CORNELL UNIV
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-02-18 至 2025-01-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10335198
• 关键词: Address; Adolescent; Affect; Affinity; Age; Antigen Presentation Pathway; Antigen-Presenting Cells; Antigens; Autoantigens; Binding; Binding Proteins; Biochemical; CD4 Positive T Lymphocytes; Cathepsins; Chronic Childhood Arthritis; Data; Detection; Diabetes Mellitus; Disease; Environment; Epitopes; Goals; Haplotypes; Hot Spot; Human; Hyperglycemia; Immune response; Immune system; Immunity; Immunization; Immunologics; In Vitro; Infection; Insulin; Insulin-Dependent Diabetes Mellitus; Interferons; Malignant Neoplasms; Maps; Mass Spectrum Analysis; Measures; Metabolic stress; Metabolic syndrome; Modification; Multiple Sclerosis; Non-Insulin-Dependent Diabetes Mellitus; Obese Mice; Oxidation-Reduction; Oxidative Stress; Oxides; Pathologic; Pathway interactions; Patients; Pattern; Peptides; Peripheral Blood Mononuclear Cell; Persons; Phenotype; Play; Population; Positioning Attribute; Post-Translational Protein Processing; Prealbumin; Process; Production; Proteins; Proteomics; Role; Salmonella typhimurium; Sampling; Stains; Surface; Surveys; System; T-Lymphocyte; TNF gene; adaptive immune response; analysis pipeline; antigen processing; autoreactive T cell; carbonyl group; chemical group; cytokine; data acquisition; diabetic patient; experimental study; glycation; high throughput analysis; immunogenicity; mouse model; neoantigens; novel; pathogen; protein aminoacid sequence; sex; sugar; tandem mass spectrometry

Approximately 32% of the population in the U.S. have metabolic syndrome, and about 10% have type 2 diabetes (T2D). In these hyperglycemic conditions proteomic modifications associated with glycation and glycoxidation are often observed. In this application we propose to analyze the role played by glycation and carbonylation on the MHC II antigen processing and presentation machinery and its implication for T cell immunity. Prompted by the observation that MHC II proteins, as well as processed peptides, are modified by advanced glycation endpoint (AGE) and carbonyl groups in patients with T2D, we aim to systematically dissect the role played by these moieties on antigen processing, MHC II-peptide binding, DM editing, and T cell presentation. In a step-by-step approach we will address how the antigen processing machinery is affected by glycation/carbonylation post-translational modifications (PTMs) and how this, in turn, affects T cell immunity. In this application we propose to: (i) map glycation and carbonylation on MHC II Molecules in T2D patients, Ob/Ob mice and relevant controls, using state-of- the-art tandem mass spectrometry, (ii) determine the effect of these PTMs on endosomal antigen processing using quantitative MS/MS and hot spot analysis, (iii) analyze the MHC-II immunopeptidomes in T2D patients, Ob/Ob mice and relevant controls to determine how the dysmetabolic environment can affect peptide selection, epitopes copy number and peptides PTM- modifications, (iv) analyze the MHC-II immunopeptidomes in Ob/Ob mice and relevant controls, prior to and after infection with S. Typhimurium, to determine how the dysmetabolic environment can affect MHC II restricted pathogen immunity, (v) characterize immune responses to novel PTM-modified autoantigens by CD4 T cell using tetramer staining following immunization with relevant epitopes, and T cell characterization (surface phenotype, proliferation and cytokine production following antigen stimulation). Ultimately our analysis will provide a mechanistic analysis of how T2D impacts MHC II- restricted immune responses and its consequences on immunity.

美国约有32％的人口患有代谢综合征，约有10％的人患有2型 糖尿病（T2D）。在这些高血糖条件下，蛋白质组学修饰与糖化和 经常观察到糖氧化。在此应用中，我们建议分析糖基化的作用 以及对MHC II抗原处理和演示机械的羰基化及其对T的影响 细胞免疫。由于观察到MHC II蛋白以及加工肽是 通过T2D患者的高级糖基化终点（年龄）和羰基修饰，我们的目标是 系统地剖析这些部分在抗原加工，MHC II肽结合中所起的作用， DM编辑和T细胞呈现。在逐步的方法中，我们将解决抗原的方式 加工机械受到糖化/羰基化修饰后（PTM）和羰基的影响 反过来，这如何影响T细胞免疫。在此应用程序中，我们建议：（i）地图糖化和 T2D患者，OB/OB小鼠和相关对照的MHC II分子的羰基化，使用最先进 ART串联质谱法，（ii）确定这些PTM对内体抗原的影响 使用定量MS/MS和热点分析处理，（III）分析MHC-II T2D患者，OB/OB小鼠和相关对照中的免疫肽组，以确定如何 差异代谢环境会影响肽选择，表位拷贝数和肽PTM- 修改，（iv）分析OB/OB小鼠中的MHC-II免疫肽和相关对照，然后 并在感染伤寒链霉菌后，以确定差异代谢环境如何影响MHC II受限的病原体免疫，（V）表征对新型PTM修饰的免疫反应 与相关表位免疫后，使用四聚体染色的CD4 T细胞自动抗原，T 细胞表征（抗原后表面表型，增殖和细胞因子产生 刺激）。最终，我们的分析将提供有关T2D如何影响MHC II-的机械分析。 限制免疫反应及其对免疫的后果。