Novel Approach to T Cell Specificity in Sjogren's Syndrome

干燥综合征 T 细胞特异性的新方法

基本信息

批准号：
8102492
负责人：
A Darise Farris
金额：
$ 41.82万
依托单位：
OKLAHOMA MEDICAL RESEARCH FOUNDATION
依托单位国家：
美国
项目类别：
财政年份：
2011
资助国家：
美国
起止时间：
2011-07-21 至 2016-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8102492
关键词：
Address Antigen Presentation Antigens Autoantibodies Autoantigens Autoimmunity Automobile Driving B-Lymphocytes Biological Response Modifier Therapy Biopsy Blood Cells Blood specimen CD4 Positive T Lymphocytes CD8-Positive T-Lymphocytes CD8B1 gene Cells Clinic Clonal Expansion Communicable Diseases Data Dendritic Cells Detection Development Disease Epitopes Evaluation Event Exocrine Glands Fractionation Frequencies Gland Haplotypes Helper-Inducer T-Lymphocyte Human Immune Immunity In Situ Infiltration Inflammatory Interferon Type I Investigation Knowledge Labial Salivary Gland Lead Lesion Lymphocyte Lymphocytic Infiltrate Malignant Neoplasms Measures Memory Minor Monitor Nuclear Antigens Oklahoma Organ Parotid Gland Pathogenesis Pathologic Patients Peripheral Positioning Attribute Production Proteins Proteomics RNA Reagent Reporter Research Research Infrastructure Research Personnel Reverse Transcriptase Polymerase Chain Reaction Role SS-A antigen Salivary Glands Screening procedure Serum Sjogren&apos s Syndrome Specificity T-Cell Antigen Receptor Specificity T-Cell Immunologic Specificity T-Cell Receptor T-Cell Receptor Genes T-Lymphocyte T-Lymphocyte Epitopes Techniques Technology Tissues Transfection Translational Research Work cell type genome wide association study innovation insight novel novel strategies peripheral blood receptor tool

项目摘要

Primary Sjogren's Syndrome (SS) is a common, debilitating disease characterized by lymphocytic infiltrates in exocrine tissue. Understanding of SS pathogenesis and development of effective biologic therapies for SS have been hampered by a lack of strategies for deciphering the specificities and differentiation states of antigen-specific T cells in the targeted tissue. We are in a unique position to address this important challenge due to i) establishment of the successful OMRF Sjogren's Research Clinic, ii) available expertise in proteomics and single cell RT-PCR of lymphocyte receptors, iii) recent discovery of HLA DR3-restricted T cell epitopes of the canonical SS autoantigens and iv) recruitment of strong collaborators who will work with us to transfer a novel T cell receptor (TCR) RNA transfection technology from the field of cancer to autoimmunity. Novel biologic therapies directed against new pathogenic T helper (Th) cell types including Th17 and T follicular helper (TFH) cells are being developed and could be applied to SS if predominant Th differentiation states in SS were known. In Aim 1 we will use a systematic single cell RT-PCR approach to characterize the TCR repertoire of CD4+ and CD8+ T cells isolated from paired labial salivary gland (SG) biopsy and peripheral blood (PB) samples of SS patients. Evaluation of these data for evidence of T cell clonal expansion and common TCR segment usage or CDRS motifs will disclose evidence for or against involvement of CD8+ T cells in SS and will identify CD4+ or CD8+ TCRs that are expanded in situ within the inflammatory lesion. In Aim 2, we will synthesize and validate DRS tetramer reagents that will detect Th cells specific for the primary known SS autoantigens Ro/SS-A and La/SS-B. These reagents will be exploited to quantify the frequency of these cells and to determine their predominant memory and Th differentiation states in SG and PB. The relationship(s) of these features to serum Type I interferon activity and measures of disease are expected to lead to insights into SS pathogenesis. In Aim S we will use a proteomic approach to identify SG antigens recognized by T cells that are clonally expanded in SG of SS patients, a strategy made feasible by the robust TCR RNA transfection technique of our collaborators.

原发性干燥综合征 (SS) 是一种常见的使人衰弱的疾病，其特征是外分泌组织中淋巴细胞浸润。由于缺乏破译靶组织中抗原特异性 T 细胞的特异性和分化状态的策略，对 SS 发病机制的理解和有效的 SS 生物疗法的开发受到阻碍。我们在应对这一重要挑战方面处于独特的地位，因为 i) 成功建立了 OMRF Sjogren 的研究诊所，ii) 拥有蛋白质组学和淋巴细胞受体单细胞 RT-PCR 方面的专业知识，iii) 最近发现了 HLA DR3 限制性 T 典型 SS 自身抗原的细胞表位，以及 iv) 招募强有力的合作者，他们将与我们合作，将新型 T 细胞受体 (TCR) RNA 转染技术从癌症领域转移到自身免疫领域。针对新的致病性 T 辅助 (Th) 细胞类型（包括 Th17 和滤泡辅助 T (TFH) 细胞）的新型生物疗法正在开发中，如果已知 SS 中的主要 Th 分化状态，则可以将其应用于 SS。在目标 1 中，我们将使用系统的单细胞 RT-PCR 方法来描述从 SS 患者的配对唇唾液腺 (SG) 活检和外周血 (PB) 样本中分离出的 CD4+ 和 CD8+ T 细胞的 TCR 库。对这些数据进行评估以获取 T 细胞克隆扩增和常见 TCR 片段使用或 CDRS 基序的证据，将揭示支持或反对 SS 中 CD8+ T 细胞参与的证据，并将识别在炎症病变内原位扩增的 CD4+ 或 CD8+ TCR。在目标 2 中，我们将合成并验证 DRS 四聚体试剂，该试剂将检测对主要已知 SS 自身抗原 Ro/SS-A 和 La/SS-B 具有特异性的 Th 细胞。这些试剂将用于量化这些细胞的频率并确定它们在 SG 和 PB 中的主要记忆和 Th 分化状态。这些特征与血清 I 型干扰素活性和疾病测量的关系预计将有助于深入了解 SS 发病机制。在 Aim S 中，我们将使用蛋白质组学方法来鉴定 SS 患者 SG 中克隆扩增的 T 细胞识别的 SG 抗原，我们合作者强大的 TCR RNA 转染技术使这一策略变得可行。