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

• 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
• 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' 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

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的淋巴细胞浸润。SS发病机制的理解和有效的SS生物疗法的发展受到阻碍，缺乏破译的特异性和抗原特异性T细胞在靶组织中的分化状态的策略。我们处于一个独特的位置，以解决这一重要的挑战，由于i）成功的OMRF Sjogren的研究诊所的建立，ii）在蛋白质组学和淋巴细胞受体的单细胞RT-PCR方面的专业知识，iii）最近发现的HLA DR 3限制性T细胞， 典型SS自身抗原的细胞表位和iv）招募强大的合作者，他们将与我们合作，将新的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。新的生物疗法针对新的致病性T辅助（Th）细胞类型，包括Th 17和T滤泡辅助（TFH）细胞正在开发中，并可能适用于SS，如果主要的Th分化状态在SS是已知的。在目标1中，我们将使用系统的单细胞RT-PCR方法， 表征分离自SS患者的成对唇唾液腺（SG）活检和外周血（PB）样品的CD 4+和CD 8 + T细胞的TCR库。对这些数据中T细胞克隆扩增和常见TCR片段使用或CDRS基序的证据进行评价，将揭示支持或反对CD 8 + T细胞参与SS的证据，并将鉴定在炎性病变内原位扩增的CD 4+或CD 8 + TCR。在目标2中，我们将合成并验证DRS四聚体试剂，其将检测对主要已知SS自身抗原Ro/SS-A和La/SS-B特异的Th细胞。这些试剂将被用来量化这些细胞的频率，并确定其主要的记忆和Th分化状态SG和PB。这些特征与血清I型干扰素活性和疾病测量的关系预期将导致对SS发病机制的深入了解。在Aim S中，我们将使用蛋白质组学方法来识别由SS患者SG中克隆扩增的T细胞识别的SG抗原，这一策略通过我们合作者的强大TCR RNA转染技术变得可行。