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: 9974042
• 负责人: LAURA SANTAMBROGIO
• 金额: $ 75.74万
• 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/9974042
• 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; 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.

在美国，大约32%的人患有代谢综合征，大约10%的人患有2型 糖尿病(T2D)。在这些高血糖条件下，与糖基化相关的蛋白质组修饰和 经常观察到糖氧化作用。在本应用中，我们建议分析糖基化所起的作用 和MHC II抗原处理和提呈机制上的羰基化及其对T细胞的意义 细胞免疫。促使观察到MHC II蛋白以及加工过的多肽 在T2D患者中，通过晚期糖基化终点(AGE)和羰基的修饰，我们的目标是 系统地剖析了这些部分在抗原加工、MHC II-肽结合、 DM编辑和T细胞演示。在一个循序渐进的方法中，我们将讨论抗原是如何 加工机械受到糖基化/羰基化翻译后修饰(PTMS)和 这反过来又是如何影响T细胞免疫的。在本申请中，我们建议：(I)映射糖基化和 T2D患者、Ob/Ob小鼠和相关对照组中MHC II分子的羰化反应 ART串联质谱学，(Ii)确定这些PTM对内体抗原的影响 利用定量MS/MS和热点分析进行处理，(III)分析MHC-II T2D患者、Ob/Ob小鼠和相关对照的免疫表位研究 代谢异常环境会影响多肽的选择、表位拷贝数和多肽PTM- 修改，(Iv)分析Ob/Ob小鼠和相关对照组的MHC-II免疫多肽，在 并在感染鼠伤寒沙门氏菌后，确定代谢障碍环境如何影响MHC II限制性病原体免疫，(V)对新型PTM修饰的免疫应答 相关表位免疫后经四聚体染色的CD4T细胞自身抗原和T细胞 细胞特性(抗原后的表面表型、增殖和细胞因子产生 刺激)。最终，我们的分析将提供T2D如何影响MHC II的机制分析- 限制性免疫反应及其对免疫的影响。