Sjogren's Syndrome Pathogenic Autoantibodies

干燥综合征致病性自身抗体

• 批准号: 10450830
• 负责人: Robert Hal Scofield
• 金额: $ 29.03万
• 依托单位: UNIVERSITY OF OKLAHOMA HLTH SCIENCES CTR
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-08-01 至 2023-02-01
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10450830
• 关键词: Address; 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 Circulation; Blood Vessels; Cell Differentiation process; Cells; Characteristics; Clinical; Data; Disease; Disease model; Dryness; Epitopes; Exocrine Glands; Functional disorder; Genomics; Gland; Hand; Harvest; Helper-Inducer T-Lymphocyte; Human; Immunoglobulin G; Immunoglobulin-Secreting Cells; Immunologics; Impairment; Individual; Joints; Kidney; Kidney Diseases; Lung; Lung diseases; Lymphocytic Infiltrate; Lymphoma; Measures; 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' 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

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.

项目摘要/摘要 Sjögren综合征（SS）是一种系统性的自身免疫性疾病，最常见的是针对外分泌 腺体的特征是持续干燥的眼睛和嘴巴以及晶状体外的参与。唾液 腺淋巴细胞浸润是该疾病的病理发现。有B细胞扩张，超级 SS患者外分泌网格中的反应性和抗体形成。一些患者有腺体生态 生发中心的形成以及II型B细胞的存在（T2）在表型上类似于边缘 脾脏中的区域（MZ）B细胞用作缺失检查点，在诱导/丢失中很重要 宽容。患者血清通常包含自身抗体（SS-A）和LA（SS-B），以及 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。其他 存在特异性，包括毒蕈碱3受体（M3R）的特异性。证据表明 靶向M3R的抗体在Para交照信号传导中很重要，可能引起腺功能障碍。我们， 其他人已经证明，受影响个体的IgG注射到幼稚的小鼠中时，可以转移 疾病表现为唾液流动障碍。这一点，证据强烈支持了b的前提 浸润SS患者唾液腺的细胞不仅会产生自身抗体，部分原因是 腺功能障碍，但也是血清中一些自身抗体的来源。该建议将测试 假设就是这种情况，并将解决该功能障碍的基础的致病机制。在 AIM 1，使用最新的尖端技术，我们将对V区进行测序并产生单克隆 来自唾液腺浆膜的抗体（mAb），并将其与在自身抗体库中进行比较 使用高分辨率轨道质谱Ig蛋白测序的同一患者的血清。 这，我们将确定循环中的抗反应是否是通过抗体分泌细胞产生的 唾液腺。鉴于抗Ro clonotypes定期翻转，确定此营业额涉及 外分泌网格中的B淋巴细胞将是对疾病发病机理的重要见解。我们 已经证明，有些患者的B细胞浸润​​了腺体，而其他患者 不要。在AIM 2中，我们将确定克隆膨胀B细胞的相关性和病理生理学 唾液腺。我们假设克隆扩展的B细胞将与其他免疫学有关 腺体中的特征，可以识别驱动生发中心的种群或微环境影响 形成，B细胞增殖和自身抗体产生。我们有绑定并阻止M3R的手动mab 激活，因此可能与疾病的发病机理有关。在AIM 3中，我们将定义 致病性mAb可以阻止唾液流动，并确定是否可以抑制它们。我们假设这一点 致病mAb将与非致病性mAb相比，将结合不同的M3R表位和 表位作为逆D-氨基酸肽的反向拟合将在我们的小鼠模型中具有阻塞作用。