Understanding the Impacts of HLA-DO in vivo

了解 HLA-DO 体内的影响

• 批准号: 9055136
• 负责人: Scheherazade Sadegh-Nasseri
• 金额: $ 40.5万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-04-01 至 2020-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9055136
• 关键词: 3' Untranslated Regions; Acute; Address; Affect; African American; Antigen Presentation Pathway; Antigens; Autoantigens; Autoimmune Diseases; B-Cell Lymphomas; B-Lymphocytes; Binding; Biological; Biological Process; CD4 Positive T Lymphocytes; Cell Count; Collaborations; Collagen; Collagen-Induced Arthritis; Complex; DR1 gene; Development; Disease; Dissociation; Drug or chemical Tissue Distribution; Epitopes; Event; Frequencies; Future; Gene Expression; Genes; Graft Rejection; HLA-DR1 Antigen; Helper-Inducer T-Lymphocyte; Histocompatibility Antigens Class II; Human; Imaging technology; Immune response; Immunization; Immunodominant Epitopes; Immunotherapeutic agent; Infection; Intervention; Kinetics; Knock-out; Knockout Mice; Lead; Ligands; Maintenance; Mediating; Molecular Chaperones; Molecular Conformation; Mus; Mutation; Nucleic Acid Regulatory Sequences; Pathway interactions; Patients; Peptide Fragments; Peptide/MHC Complex; Peptides; Phenotype; Play; Predisposition; Process; Production; Proteins; Recombinants; Regulation; Reporting; Resistance; Rheumatoid Arthritis; Risk; Role; Sampling; Self Tolerance; Single Nucleotide Polymorphism; Staining method; Stains; T-Lymphocyte; Testing; Therapeutic; Thymus Gland; Transgenic Mice; Untranslated Regions; Work; adaptive immunity; antigen processing; autoreactive T cell; biophysical analysis; co-infection; density; design; dimer; fighting; fluorescence imaging; genome wide association study; improved; in vivo; inhibitor/antagonist; mouse model; mutant; novel; novel therapeutics; overexpression; pathogen; peptide A; prevent; receptor; tumor

Project Title: Understanding the impacts of HLA-DO in vivo The project described in this R01 application addresses a cellular study aimed at verification of mechanistic understanding of the regulatory role HLA-DO (H2-O in mice) (DO), an accessory molecule of antigen processing and presentation pathway that is dependent on HLA-DM (H2-M in mice) on its cellular expression and has restricted tissue distribution. Despite the discovery of DO for over two decades ago, an understanding of its impact in regulation of antigen processing and epitope selection has been lacking. The current understanding of the DO function is that it inhibits DM. Recently, we have demonstrated that DO does not inhibit DM: we have shown that DO interacts with human MHC class II, HLA-DR1 (DR1), molecules directly and in collaboration with DM optimizes epitope selection. We demonstrated that DO has differential effects on binding of different peptides to DR1 molecules. Those findings need in vivo verification. The key question addressed here is whether DO plays a role in regulation of epitope selection in the thymus leading to changing the expressed T cell repertoire, and possibly regulation of susceptibility to the development of autoimmune diseases. In aim I, we would examine the role of DO in altering T cell repertoire in DR1 expressing transgenic mice that do, or do not express H2-O. Using unique mouse models expressing a mutant DR1 (DR1bG86Y) that interacts with DO but not with DM, with or without H2-O. In Aim 2, we would address whether DO inhibits DM, or its function is different from inhibiting DM in vivo. In Aim 3a, we would examine precursor frequency for the dominant disease associated epitope of collagen II, the causative antigen in Collagen Induced Arthritis (CIA), in in DR1+ DO-knockout mice. In Aim3b we would find out susceptibility of DO-KO mice to CIA, using novel non-invasive fluorescent imaging technology developed by our colleagues at JHU. In Aim 3c we would examine samples from Rheumatoid Arthritis patients that are identified as having a single nucleotide polymorphism (SNP) in their HLA-DOA 3'UTR gene for the expression of HLA-DO by intracellular FACS staining. The in vivo verification of HLA-DO contribution to the regulation of antigen processing and epitope selection would be a major leap towards filling the gap in understanding the biological significance of HLA-DO, which can guide the design of effective immunotherapeutics in future.

项目名称：了解HLA-DO在体内的影响 本R 01申请中描述的项目涉及一项细胞研究，旨在验证 了解抗原辅助分子HLA-DO（H2-O in mice）（DO）的调节作用 依赖于HLA-DM（小鼠中的H2-M）对其细胞表达的加工和呈递途径 并且具有受限的组织分布。尽管DO的发现是在二十多年前， 其在调节抗原加工和表位选择中的影响一直缺乏。当前 对DO功能的理解是它抑制DM。最近，我们已经证明DO不会 抑制DM：我们已经证明DO与人MHC II类分子HLA-DR 1（DR 1）直接相互作用 并与DM合作优化表位选择。我们证明了DO对 不同肽与DR 1分子的结合。这些发现需要在体内验证。关键问题 这里讨论的是DO是否在胸腺中表位选择的调节中起作用，从而导致改变 表达的T细胞库，并可能调节自身免疫性疾病的发生， 疾病在目的I中，我们将检查DO在改变表达DR 1的转基因小鼠中的T细胞库中的作用。 表达或不表达H2-O的小鼠。使用表达突变型DR 1（DR 1bG 86 Y）的独特小鼠模型 与DO相互作用但不与DM相互作用，有或没有H2-O。在目标2中，我们将讨论DO是否抑制 DM，或其功能不同于体内抑制DM。在目标3a中，我们将检查 II型胶原的主要疾病相关表位，胶原诱导性关节炎的致病抗原 (CIA)在DR 1 + DO敲除小鼠中。在Aim 3b中，我们将发现DO-KO小鼠对CIA的易感性，使用 JHU同事开发的新型非侵入性荧光成像技术。在目标3c中，我们将 检查来自风湿性关节炎患者的样本，这些患者被鉴定为具有单核苷酸 通过细胞内流式细胞术检测HLA-DOA 3 'UTR基因中的SNP对HLA-DO表达的影响 染色HLA-DO对抗原加工和表位调节的体内验证 选择将是在理解HLA-DO的生物学意义上填补差距的一个重大飞跃， 这可以指导未来有效的免疫治疗剂的设计。