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.

大约 32% 的美国人口患有代谢综合征，其中约 10% 患有 2 型代谢综合征 糖尿病（T2D）。在这些高血糖条件下，与糖化和相关的蛋白质组修饰 经常观察到糖氧化作用。在此应用中，我们建议分析糖化所发挥的作用 MHC II 抗原加工和呈递机制中的羰基化及其对 T 的影响 细胞免疫。观察到 MHC II 蛋白以及经过加工的肽， 通过 T2D 患者的晚期糖基化终点 (AGE) 和羰基进行修改，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、 DM 编辑和 T 细胞呈现。我们将逐步解决抗原如何 加工机械受糖化/羰基化翻译后修饰 (PTM) 的影响， 这又如何影响 T 细胞免疫。在此应用中，我们建议：（i）绘制糖化图谱和 T2D 患者、Ob/Ob 小鼠和相关对照中 MHC II 分子的羰基化，使用状态- 最先进的串联质谱法，(ii) 确定这些 PTM 对内体抗原的影响 使用定量 MS/MS 和热点分析进行处理，(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 代谢异常环境会影响肽选择、表位拷贝数和肽 PTM- 修改，(iv) 分析 Ob/Ob 小鼠和相关对照中的 MHC-II 免疫肽组，然后 感染鼠伤寒沙门氏菌后，确定代谢异常环境如何影响 MHC II 限制性病原体免疫，(v) 表征对新型 PTM 修饰的免疫反应 使用相关表位免疫后使用四聚体染色的 CD4 T 细胞产生自身抗原，并且 T 细胞特征（表面表型、增殖和抗原后细胞因子的产生 刺激）。最终，我们的分析将提供 T2D 如何影响 MHC II 的机制分析 - 免疫反应受限及其对免疫力的影响。