Understanding the Impacts of HLA-DO in vivo

了解 HLA-DO 体内的影响

基本信息

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
项目类别：
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&apos 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

项目摘要

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在体内的影响此R01应用程序中描述的项目介绍了一项旨在验证机械的细胞研究了解调节作用HLA-DO（小鼠H2-O）（DO），抗原的辅助分子依赖于HLA-DM（小鼠中的H2-M）在其细胞表达上的处理和呈现途径并具有限制的组织分布。尽管二十多年前发现了DO，但缺乏对抗原加工和表位选择的影响的影响。电流对DO功能的理解是它抑制了DM。最近，我们证明了确实没有抑制DM：我们已经证明确实与人类MHC II类，HLA-DR1（DR1），分子直接相互作用并与DM合作优化了表位选择。我们证明DO对不同肽与DR1分子的结合。这些发现需要体内验证。关键问题这里解决表达的T细胞曲目，并可能调节自身免疫性发展的易感性疾病。在AIM I中，我们将研究DO在DR1中改变T细胞库中的作用，以表达转基因做或不表达H2-O的小鼠。使用表达突变体DR1（DR1BG86Y）的独特鼠标模型与DM相互作用，但没有H2-O。在AIM 2中，我们将解决是否抑制 DM或其功能不同于抑制体内DM。在AIM 3A中，我们将检查前体频率胶原蛋白II的主要疾病相关的表位，胶原蛋白诱导关节炎的致病抗原（CIA），在DR1+ DO-DO-KNOCKOUT小鼠中。在AIM3B中，我们会发现Do-Ko小鼠对CIA的敏感性，新型的非侵入性荧光成像技术由JHU的同事开发。在AIM 3C中，我们会检查来自类风湿关节炎患者的样品，这些样本被鉴定为具有单核苷酸其HLA-DOA 3'UTR基因中的多态性（SNP），用于细胞内FACS的HLA-DO表达染色。 HLA-DO对抗原加工和表位调节的贡献的体内验证选择将是填补HLA-DO的生物学意义的空白的重大飞跃，这可以指导将来有效的免疫治疗剂设计。