CD4+ T-cell Repertoires of the Lung in Rheumatoid Arthritis

类风湿性关节炎肺部 CD4 T 细胞库

• 批准号: 10493368
• 负责人: SHAODONG DAI
• 金额: $ 18.98万
• 依托单位: UNIVERSITY OF COLORADO DENVER
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-22 至 2024-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10493368
• 关键词: Antibodies; Antibody Formation; Antigens; Autoantigens; Autoimmune Diseases; Autoimmune Responses; Autoimmunity; B-Lymphocytes; Biological Assay; Blood; CD4 Positive T Lymphocytes; Cells; Clinical; Clonal Expansion; Complex; Data; Data Set; Detection; Development; Disease; Environment; Epitope spreading; Etiology; Exposure to; Foundations; Frequencies; Gene Expression; Genetic Diseases; Genomics; Glycine; Goals; Heavy Metals; Heterogeneity; Hydrophobicity; Individual; Inflammatory; Inflammatory Arthritis; Interleukin-17; Interleukin-2; Interstitial Lung Diseases; Joints; Knowledge; Lead; Lesion; Location; Lung; Lung diseases; Lymphocyte; Measures; Microfluidic Microchips; Microfluidics; Monitor; Pathogenesis; Pathogenicity; Pathologic; Patients; Peptides; Peripheral; Phenotype; Pollution; Population; Publishing; Recovery; Research; Rheumatoid Arthritis; Risk; Role; Sampling; Serum; Silicon Dioxide; Site; Smoking; Specificity; Sputum; Stains; Stimulus; Surface; Synovial Fluid; Synovial Membrane; Synovitis; T cell receptor repertoire sequencing; T cell response; T-Cell Receptor; T-Lymphocyte; T-Lymphocyte Subsets; TNFRSF10A gene; Techniques; Testing; Therapeutic; Touch sensation; adaptive immune response; antigen detection; autoimmune arthritis; autoreactivity; base; biomarker discovery; citrullinated protein; cohort; complementarity-determining region 3; cytokine; cytotoxic; improved; interest; joint inflammation; joint injury; mucosal site; novel; particle; pathogen; peripheral blood; personalized medicine; response; single-cell RNA sequencing; systemic autoimmune disease; transcriptome sequencing; Antibodies; Antibody Formation; Antigens; Autoantigens; Autoimmune Diseases; Autoimmune Responses; Autoimmunity; B-Lymphocytes; Biological Assay; Blood; CD4 Positive T Lymphocytes; Cells; Clinical; Clonal Expansion; Complex; Data; Data Set; Detection; Development; Disease; Environment; Epitope spreading; Etiology; Exposure to; Foundations; Frequencies; Gene Expression; Genetic Diseases; Genomics; Glycine; Goals; Heavy Metals; Heterogeneity; Hydrophobicity; Individual; Inflammatory; Inflammatory Arthritis; Interleukin-17; Interleukin-2; Interstitial Lung Diseases; Joints; Knowledge; Lead; Lesion; Location; Lung; Lung diseases; Lymphocyte; Measures; Microfluidic Microchips; Microfluidics; Monitor; Pathogenesis; Pathogenicity; Pathologic; Patients; Peptides; Peripheral; Phenotype; Pollution; Population; Publishing; Recovery; Research; Rheumatoid Arthritis; Risk; Role; Sampling; Serum; Silicon Dioxide; Site; Smoking; Specificity; Sputum; Stains; Stimulus; Surface; Synovial Fluid; Synovial Membrane; Synovitis; T cell receptor repertoire sequencing; T cell response; T-Cell Receptor; T-Lymphocyte; T-Lymphocyte Subsets; TNFRSF10A gene; Techniques; Testing; Therapeutic; Touch sensation; adaptive immune response; antigen detection; autoimmune arthritis; autoreactivity; base; biomarker discovery; citrullinated protein; cohort; complementarity-determining region 3; cytokine; cytotoxic; improved; interest; joint inflammation; joint injury; mucosal site; novel; particle; pathogen; peripheral blood; personalized medicine; response; single-cell RNA sequencing; systemic autoimmune disease; transcriptome sequencing

PROJECT SUMMARY Rheumatoid arthritis (RA) is an inflammatory arthritis of autoimmune origin leading to erosive joint damage. While B-cell involvement in RA is well characterized by antibodies to citrullinated protein antigens (ACPAs), much less is known about T-cell reactivity and functionality in the etiology and pathogenesis of RA. Studies have focused largely on the T-cells of the joint and peripheral blood. However, the joint is reflective of long-standing autoimmunity which introduces a bias in adaptive immune responses due to epitope spreading. ACPAs exist systemically and in mucosal sites preceding the onset of joint inflammation, which suggests the originating site of autoimmunity in RA is extra-articular. The lung is one of these sites considered to be a possible location for initiating RA-related autoimmunity. Notably, sputum ACPAs are detectable in over 33% of those at risk for RA and over 66% of those with RA. Furthermore, lung abnormalities are seen in over 70% of those with RA, and lung disease is a common extra-articular manifestation of RA. The goal of this proposal is to characterize the T- cell repertoire of the lung in RA to better understand disease pathogenesis. This involves investigating both phenotype and functionality of RA-related lung T-cells through single cell RNA-seq as well as the antigenic reactivity of these T-cells through IL-2 bioassays when presented with synovial fluid antigens. Studying the T- cell responses of the lung has been a challenge due to sampling limitations. Induced sputum is an inexpensive, non-invasive sample but it typically contains 1% or fewer lymphocytes. We have overcome the limitations of sputum for studying T-cell responses of the lung by the use of a novel microfluidic technique to recover sputum lymphocytes in a completely touch-free and unbiased manner. This allows us to study the T-cells of the lung in RA and compare them to the RA synovium as we hypothesize there will be shared effector states and antigenic reactivity between lung-derived and synovium-derived CD4+ T-cells in RA. Our overall hypothesis is T-cells derived from the lung have common effector states and antigenic reactivity to the T-cell repertoire of the RA synovium. We will assess sputum lymphocytes in RA for the presence of Vβ17+ peripheral helper T (Tph) and Vβ14+ cytotoxic CD4+ T-cells and compare T-cell receptor motifs and antigenic reactivity of T-cells in RA sputum to RA synovium. From the single-cell RNA-seq data, we will also obtain TCR sequences of the RA sputum CD4+ T-cells. We will use previously published datasets containing TCR α and β chain sequences to determine closely related TCRs and motifs between the RA synovium and lung. The findings from our proposal will allow for an improved understanding of T-cell involvement in RA in both the lung as well as the joint.

项目摘要 类风湿关节炎（RA）是自身免疫起源的炎症性关节炎，导致侵蚀性关节损伤。尽管 B细胞参与RA的特征是与瓜氨酸蛋白抗原（ACPA）的抗体，更不用说 关于RA病因和发病机理中T细胞反应性和功能的已知。研究重点是 主要在关节和外周血的T细胞上。但是，该关节反映了长期存在的 自身免疫性引入了由于表位扩散而导致自适应免疫调查的偏见。 ACPA存在 在关节炎症开始之前的系统和粘膜部位，这表明原始部位 RA自身免疫性是外关节的。肺是这些地点之一，被认为是 引发了与RA相关的自身免疫。值得注意的是，有超过33％有RA的风险的痰液可检测到痰液ACPA 以及有66％以上的RA。此外，在70％的患有RA和 肺部病是RA的常见关节外表现。该提议的目的是表征t- RA中肺的细胞曲目，以更好地了解疾病发病机理。这涉及研究两者 通过单细胞RNA-Seq以及抗原的RA相关肺T细胞的表型和功能 当用滑液抗原呈现这些T细胞通过IL-2生物测定的反应性。研究T细胞 由于采样限制，肺部的细胞反应一直是一个挑战。诱导的痰是便宜的， 非侵入性样本，但通常包含1％或更少的淋巴细胞。我们已经克服了 通过使用新型微流体技术来恢复痰液，用于研究肺的T细胞反应的痰 以完全无触摸和无偏的方式进行淋巴细胞。这使我们能够研究肺的T细胞 RA并将它们与RA Synovium进行比较，因为我们假设会有共享效应态和抗原 RA中的肺衍生和滑膜衍生的CD4+ T细胞之间的反应性。我们的总体假设是T细胞 源自肺的效应态和对RA的T细胞库的抗原反应性的抗原反应性 滑膜。我们将评估RA中的痰液淋巴细胞，以了解Vβ17+外围辅助辅助剂T（TPH）和 Vβ14+细胞毒性CD4+ T细胞，并比较ra痰中T细胞的T细胞受体基序和抗原反应性 到RA滑膜。从单细胞RNA-seq数据中，我们还将获得RA痰CD4+的TCR序列 T细胞。我们将使用先前发表的包含TCRα和β链序列的数据集来确定 RA滑膜和肺之间的相关TCR和基序。我们提案的发现将允许 对肺和关节的RA中T细胞参与的了解得以提高。