Sjogren's Syndrome Pathogenic Autoantibodies

干燥综合征致病性自身抗体

基本信息

批准号：
10854472
负责人：
Robert Hal Scofield
金额：
$ 29.1万
依托单位：
OKLAHOMA MEDICAL RESEARCH FOUNDATION
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-08-01 至 2024-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10854472
关键词：
Affect Amino Acids Animal Model Antibodies Antibody Formation Antibody titer measurement Autoantibodies Autoantigens Autoimmune Diseases B Cell Proliferation B-Cell Antigen Receptor B-Lymphocytes Binding Blocking Antibodies Blood Blood Vessels Cell Differentiation process Cells Characteristics Circulation Clinical Clonal Expansion Data Disease Disease model Dryness Epitopes Exocrine Glands Functional disorder Genomics Gland Hand Harvest Helper-Inducer T-Lymphocyte Hematological Disease Human Immunoglobulin G Immunoglobulin-Secreting Cells Immunologics Impairment Individual Infiltration Joints Kidney Kidney Diseases Lung Lung diseases Lymphocytic Infiltrate Lymphoma Measures Membrane Methods Minor salivary gland structure Monoclonal Antibodies Mus Muscarinic Acetylcholine Receptor Muscarinics Nature Pathogenesis Pathogenicity Pathologic Patients Peptide Sequence Determination Peptides Peripheral Phenotype Plasmablast Population Production Proteomics Receptor Activation Recombinants Research Resolution Risk Saliva Salivary Salivary Glands Serum Signal Transduction Sjogren&apos s Syndrome Skin Source Specificity Spleen Structure of germinal center of lymph node Symptoms System T memory cell T-Lymphocyte Techniques Testing Tissues Xerostomia arthropathies autoreactive B cell eye dryness human disease inhibiting antibody innovation insight mouse model novel therapeutics pathogenic autoantibodies peripheral blood receptor saliva secretion skin disorder systemic autoimmune disease

项目摘要

Project Summary/Abstract Sjögren's syndrome (SS) is a systemic autoimmune disease that most commonly targets the exocrine glands and is characterized by persistent dry eyes and mouth as well as extra-glandular involvement. Salivary gland lymphocytic infiltrates are a pathological finding in the disease. There are B cell expansions, hyper- reactivity and antibody formation in exocrine glands of SS patients. Some patients have glandular ectopic germinal center formation, as well as the presence of Type II B cells (T2) phenotypically similar to marginal zone (MZ) B cells in the spleen serving as a checkpoint for deletion and are important in the induction/loss of tolerance. Patient serum commonly contains autoantibodies to Ro (SS-A) and La (SS-B), and the number of anti-Ro and -La specific B cells in salivary infiltrates correlate with antibody titer in the serum. Other specificities are present, including those towards muscarinic 3 receptor (M3R). The evidence suggests antibodies targeting M3R, important in para-sympathetic signaling, may induce glandular dysfunction. We, and others have demonstrated that IgG from affected individuals, when injected into naïve mice, can transfer disease as manifested by salivary flow impairment. Thus, the evidence strongly supports the premise that B cells infiltrating the salivary glands of SS patients not only make autoantibodies that are in part responsible for glandular dysfunction but are also a source of some autoantibodies in the sera. This proposal will test the hypothesis that this is the case, and will address the pathogenic mechanisms underlying this dysfunction. In Aim 1, using the latest cutting-edge techniques, we will sequence the V-regions and produce monoclonal antibodies (mAb) from salivary gland plasmablasts, and compare them to the autoantibody repertoire found in the serum of the same patients, using high-resolution Orbitrap mass spectrometric Ig protein sequencing. Thus, we will determine whether anti-Ro in the circulation is produced by antibody-secreting cells in the salivary glands. Given that anti-Ro clonotypes are turned over regularly, determining that this turnover involves the B lymphocytes in the exocrine glands will be an important insight into the pathogenesis of the disease. We have demonstrated that some patients have clonally expanded B cells infiltrating the gland, while other patients do not. In Aim 2 we will determine the correlates and pathophysiology of clonally expanded B cells infiltrating the salivary glands. We hypothesize that clonally expanded B cells will be related to other immunological features in the gland and may identify populations or microenvironmental influences that drive germinal center formation, B cell proliferation and autoantibody production. We have in hand SS mAb that bind and block M3R activation, and thus, may be involved in the pathogenesis of the disease. In Aim 3 we will define the nature of pathogenic mAbs that block salivary flow, and determine if they can be inhibited. We hypothesize that pathogenic mAb will bind distinct M3R epitopes compared to non-pathogenic mAb and representation of the epitope as an inverse D-amino acid peptide will have a blocking action in our mouse model.

项目概要/摘要干燥综合征 (SS) 是一种全身性自身免疫性疾病，最常见的目标是外分泌腺体，其特征是持续的眼睛和口腔干涩以及腺外受累。腺体淋巴细胞浸润是该疾病的病理发现，有 B 细胞扩张、过度增殖。 SS 患者外分泌腺的反应性和抗体形成。一些患者存在腺体异位。生发中心的形成，以及表型与边缘细胞相似的 II 型 B 细胞 (T2) 的存在脾脏中的区 (MZ) B 细胞作为删除的检查点，对于诱导/丢失很重要患者血清通常含有 Ro (SS-A) 和 La (SS-B) 自身抗体，以及其数量。唾液浸润中的抗 Ro 和 La 特异性 B 细胞与血清中的抗体滴度相关。证据表明存在特异性，包括针对毒蕈碱 3 受体 (M3R) 的特异性。针对副交感神经信号转导很重要的 M3R 抗体可能会导致腺体功能障碍。其他人已经证明，将受影响个体的 IgG 注射到幼鼠体内后，可以将表现为唾液流障碍的疾病因此，证据强烈支持 B 的前提。浸润 SS 患者唾液腺的细胞不仅产生自身抗体，而自身抗体在一定程度上负责腺体功能障碍，但也是血清中一些自身抗体的来源。该提案将测试假设情况确实如此，并将解决这种功能障碍背后的致病机制。目标1，利用最新尖端技术，对V区进行测序并生产单克隆抗体来自唾液腺浆母细胞的抗体（mAb），并将它们与在体内发现的自身抗体库进行比较使用高分辨率 Orbitrap 质谱 Ig 蛋白测序对同一患者的血清进行分析。因此，我们将确定循环中的抗Ro是否是由体内的抗体分泌细胞产生的。鉴于抗 Ro 克隆型定期更新，确定这种更新涉及外分泌腺中的 B 淋巴细胞将有助于了解该疾病的发病机制。已经证明，一些患者的 B 细胞克隆性扩增，浸润腺体，而其他患者在目标 2 中，我们将确定克隆扩增的 B 细胞浸润的相关性和病理生理学。我们勇敢地相信，克隆扩增的 B 细胞将与其他免疫学相关。腺体的特征，可以识别驱动生发中心的种群或微环境影响我们拥有结合并阻断 M3R 的 SS mAb。激活，因此可能参与疾病的发病机制，在目标 3 中，我们将定义疾病的本质。阻断唾液流动的致病性单克隆抗体，并确定它们是否可以被抑制。与非致病性 mAb 相比，致病性 mAb 将结合不同的 M3R 表位，并且代表作为反 D-氨基酸肽的表位将在我们的小鼠模型中具有阻断作用。