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

类风湿性关节炎肺部 CD4 T 细胞库

基本信息

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
项目类别：
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

项目摘要

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.

项目概要类风湿性关节炎（RA）是一种自身免疫性炎症性关节炎，可导致糜烂性关节损伤。 B 细胞参与 RA 的特点是针对瓜氨酸蛋白抗原 (ACPA) 的抗体，更不用说已知 T 细胞的反应性和功能在 RA 的病因学和发病机制中的作用。很大程度上取决于关节的T细胞和外周血，然而，关节是长期存在的反映。由于 ACPA 的存在，自身免疫会导致适应性免疫反应出现偏差。在关节炎症发生之前全身和粘膜部位，这表明起源部位 RA 中自身免疫的发生是关节外的，肺是被认为可能发生的部位之一。引发 RA 相关的自身免疫值得注意的是，在 33% 以上的 RA 风险人群中可检测到痰液 ACPA。超过 66% 的 RA 患者此外，超过 70% 的 RA 患者出现肺部异常。肺部疾病是 RA 的常见关节外表现，本提案的目标是描述 T- 的特征。 RA 中肺部的细胞库，以更好地了解疾病发病机制，这涉及到对两者的研究。通过单细胞 RNA-seq 以及抗原分析 RA 相关肺 T 细胞的表型和功能研究 T 细胞在接触滑液抗原时通过 IL-2 生物测定来检测这些 T 细胞的反应性。由于采样限制，肺部细胞反应一直是一个挑战。非侵入性样本，但它通常含有 1% 或更少的淋巴细胞，我们已经克服了这些限制。使用新型微流体技术回收痰液来研究肺部 T 细胞反应淋巴细胞以完全无接触且公正的方式进行研究，这使我们能够研究肺部的 T 细胞。 RA 并将它们与 RA 滑膜进行比较，正如我们所接受的那样，它们将具有共享的效应器状态和抗原 RA 中肺源性和滑膜源性 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 RA 的 T 细胞库具有共同的效应子状态和抗原反应性我们将评估 RA 痰淋巴细胞中是否存在 Vβ17+ 外周辅助 T (Tph) 和 Vβ14+ 细胞毒性 CD4+ T 细胞并比较 RA 痰中 T 细胞受体基序和 T 细胞的抗原反应性从单细胞RNA-seq数据中，我们还将获得RA痰液CD4+的TCR序列。我们将使用先前发布的包含 TCR α 和 β 链序列的数据集来仔细确定。 RA 滑膜和肺之间相关的 TCR 和基序我们提案的发现将允许提高了对 T 细胞参与肺部和关节 RA 的了解。