Anti-Jo-1 Immune Responses in Autoimmune Myositis

自身免疫性肌炎中的抗 Jo-1 免疫反应

• 批准号: 7116917
• 负责人: STUART M LEVINE
• 金额: $ 12.9万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-08-27 至 2009-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7116917
• 关键词: HeLa cells; MHC class I antigen; aminoacid tRNA ligase; autoantigens; autoimmune disorder; clinical research; cytotoxic T lymphocyte; flow cytometry; human subject; immune response; immunocytochemistry; lung; molecular site; muscle; myositis; protein localization; protein structure function; serine proteinases

DESCRIPTION (provided by applicant): The primary objective of this mentored clinical scientist development award is to acquire the skills and expertise needed to become an independent investigator in the rheumatic diseases. This will occur through a period of intensive didactic and research training in the Division of Rheumatology at the Johns Hopkins University School of Medicine. The scientific objective of this proposal is to define the mechanisms underlying the specific immune response to histidyl-tRNA synthetase (HRS, Jo-1) in patients with autoimmune myositis and interstitial lung disease. It is known that the majority of myositis-specific autoantigens are unified by their susceptibility to cleavage by the cytotoxic lymphocyte protease granzyme B (GrB). However, the relevance of this unique property of autoantigens remains unknown. We propose that (i) the T cell response to HRS is directed at the GrB cleavage site, and (ii) that conformation and cleavability of HRS by GrB is altered in the tissue in which the autoimmune response is initiated and propagated. This proposal will address these hypotheses in Jo-1-positive patients with autoimmune myositis and interstitial lung disease, through the following specific aims: 1. To study the role of the HRS grB cleavage site in defining its immunodominant CD4+ T cell epitope. T cells from myositis patients will be probed for anti-HRS responses in-vitro using purified protein and peptides that span the GrB cleavage site. 2. To identify whether the CD8+ cytotoxic T cells in myositis patients are HRS-specific. A predictive approach will be used to identify a set of HLA-restricted HRS peptides that react with specific cytotoxic lymphocytes in myositis patients, and CD8+ T cell lines will be assessed for their ability to lyse specific target cells. 3. To determine whether an immunogenic (grB- cleavable) conformation of HRS is differentially expressed in muscle and lung tissue in patients with myositis/ILD. Lung and muscle tissue samples from patients with myositis/ILD will be probed using novel antibody reagents that recognize the GrB cleavage site. These studies will provide important information on the role of the GrB cleavage site and/or its cleavage in the immune response to HRS, as well as on tissue-specific changes in autoantigen structure that might account for its targeting in myositis/interstitial lung disease.

描述（由申请人提供）： 这个受过指导的临床科学家发展奖的主要目标是获得成为风湿病独立研究者所需的技能和专业知识。这将通过约翰·霍普金斯大学医学院风湿病学的一段密集的教学和研究培训进行。该提案的科学目标是定义对自身免疫性肌炎和间质性肺部疾病患者中对组酰基-TRNA合成酶（HRS，JO-1）的特定免疫反应的基础机制。众所周知，大多数特异性肌炎特异性自身抗原因其易感性而被细胞毒性淋巴细胞蛋白酶颗粒粒B（GRB）统一。但是，这种自动抗原的独特属性的相关性仍然未知。我们提出（i）T细胞对HRS的响应针对GRB裂解位点，（ii）在启动和传播自身免疫反应的组织中，GRB对HRS的构象和裂解性发生了改变。该建议将通过以下特定目的解决JO-1阳性自身免疫性肌炎和间质肺疾病的患者中的这些假设：1。研究HRS GRB裂解部位在确定其免疫鼻CD4+ T细胞表位中的作用。肌炎患者的T细胞将使用跨GRB裂解位点的纯化蛋白质和肽进行抗HRS反应探测。 2。确定肌炎患者中的CD8+细胞毒性T细胞是否特异性HRS。一种预测方法将用于识别一组HLA限制的HRS肽，该肽与肌炎患者中的特定细胞毒性淋巴细胞反应，并且将评估CD8+ T细胞系的CD8+ T细胞系的能力。 3。确定HRS的免疫原性（可裂解）构象是否在肌炎/ILD患者的肌肉和肺组织中差异表达。将使用识别GRB裂解部位的新型抗体试剂探测来自肌炎/ILD患者的肺和肌肉组织样品。这些研究将提供有关GRB裂解位点和/或裂解在对HRS免疫反应中的作用的重要信息，以及自身抗原结构的组织特异性变化，这可能解释其靶向肌炎/间质肺疾病。