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型代谢综合征。 糖尿病（T2 D）。在这些高血糖状况中，与糖化相关的蛋白质组学修饰， 经常观察到糖氧化。在本申请中，我们提出分析糖化所起的作用， 和羰基化对MHCII抗原加工和呈递机制的影响及其对T 细胞免疫通过观察到MHC II蛋白质以及加工的肽， 在T2 D患者中通过晚期糖基化终点（AGE）和羰基修饰，我们的目的是 系统地剖析这些部分在抗原加工，MHC II-肽结合， DM编辑和T细胞呈递。在一步一步的方法，我们将解决如何抗原 糖基化/羰基化翻译后修饰（PTM）影响加工机制， 这又是如何影响T细胞免疫力的在本申请中，我们提出：（i）映射糖化和 在T2 D患者、Ob/Ob小鼠和相关对照中，使用 最先进的串联质谱，（ii）确定这些PTM对内体抗原的影响 使用定量MS/MS和热点分析进行处理，（iii）分析MHC-II T2 D患者、Ob/Ob小鼠和相关对照中的免疫肽组，以确定T2 D患者、Ob/Ob小鼠和相关对照中的免疫肽组如何表达。 代谢异常的环境可以影响肽的选择、表位拷贝数和肽PTM。 （iv）分析Ob/Ob小鼠和相关对照中的MHC-II免疫肽组， 感染S.鼠伤寒沙门氏菌，以确定代谢异常的环境如何影响MHC II限制的病原体免疫，（V）表征对新的PTM修饰的 在用相关表位免疫后，使用四聚体染色通过CD 4 T细胞的自身抗原，和T 细胞表征（表面表型、增殖和抗原诱导后的细胞因子产生 刺激）。最终，我们的分析将提供T2 D如何影响MHC II的机制分析。 限制免疫反应及其对免疫力的影响。