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Sjogren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的淋巴细胞浸润。缺乏破译目标组织中抗原特异性T细胞的特异性和分化状态的策略，对SS发病机理的理解和有效生物疗法的发展受到了阻碍。由于i）建立成功的OMRF Sjogren研究诊所，ii）蛋白质组学和淋巴细胞受体单细胞RT-PCR的可用专业知识，iii），最近发现HLA DR3限制性T，我们处于独特的立场来应对这一重要挑战。规范SS自动抗原和IV的细胞表位）招募了强大的合作者，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性。针对新的病原T辅助辅助器（TH）细胞类型（包括Th17和T卵泡辅助（TFH）细胞）的新型生物疗法正在开发，如果已知SS中的主要TH分化态，则可以应用于SS。在AIM 1中，我们将使用系统的单细胞RT-PCR方法进行表征了SS患者的CD4+和CD8+ T细胞的TCR曲目从成对的唇唾液腺（SG）活检和外周血（PB）样品中分离出来的CD4+和CD8+ T细胞。评估这些数据，以证明T细胞克隆扩张和常见的TCR段使用或CDRS图案的证据将披露CD8+ T细胞在SS中的参与或反对SS的证据，并将识别炎症性病变内局部扩展的CD4+或CD8+ TCR。在AIM 2中，我们将合成并验证DRS四聚体试剂，该试剂将检测到针对主要已知SS自动抗原RO/SS-A和LA/SS-B的TH细胞。这些试剂将被利用以量化这些细胞的频率，并确定其在SG和PB中的主要记忆和Th分化状态。这些特征与血清I型干扰素活性的关系和疾病的度量有望导致对SS发病机理的见解。在AIM中，我们将使用一种蛋白质组学方法来识别由T细胞识别的SG抗原，这些抗原在SS患者的SG中被克隆扩展，这是通过合作者强大的TCR RNA转染技术可行的策略。