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 体内的影响 R01 申请中描述的项目涉及一项细胞研究，旨在验证机制了解抗原辅助分子 HLA-DO（小鼠中的 H2-O）(DO) 的调节作用依赖于 HLA-DM（小鼠中的 H2-M）细胞表达的加工和呈递途径并且组织分布受限。尽管溶解氧在二十多年前就被发现了，但人们对溶解氧的认识还不够深入。其对抗原加工和表位选择调节的影响尚不清楚。目前的对 DO 功能的理解是它抑制 DM。最近，我们已经证明 DO 并不抑制 DM：我们已经证明 DO 与人类 MHC II 类、HLA-DR1 (DR1) 分子直接相互作用并与 DM 合作优化表位选择。我们证明了 DO 对不同肽与 DR1 分子的结合。这些发现需要体内验证。关键问题这里要解决的是 DO 是否在调节胸腺表位选择中发挥作用，从而导致改变表达的 T 细胞库，以及对自身免疫发展的易感性的可能调节疾病。在目标 I 中，我们将检查 DO 在改变表达 DR1 的转基因 T 细胞库中的作用表达或不表达 H2-O 的小鼠。使用表达突变体 DR1 (DR1bG86Y) 的独特小鼠模型与 DO 相互作用，但不与 DM 相互作用，无论有或没有 H2-O。在目标 2 中，我们将解决 DO 是否抑制 DM，或其功能不同于体内抑制DM。在目标 3a 中，我们将检查前兆频率胶原蛋白 II 的主要疾病相关表位，胶原蛋白诱导关节炎的致病抗原 (CIA)，在 DR1+ DO 敲除小鼠中。在 Aim3b 中，我们将使用以下方法找出 DO-KO 小鼠对 CIA 的敏感性由我们在约翰霍普金斯大学的同事开发的新型非侵入性荧光成像技术。在 Aim 3c 中，我们会检查类风湿关节炎患者的样本，这些样本被鉴定为具有单核苷酸 HLA-DOA 3'UTR 基因中的多态性 (SNP) 用于通过细胞内 FACS 表达 HLA-DO 染色。 HLA-DO对抗原加工和表位调节的体内验证选择将是填补 HLA-DO 生物学意义理解空白的重大飞跃，这可以指导未来有效免疫治疗的设计。