B Cells as Antigen Presenting Cells in Human Autoimmune Diseases

B 细胞作为人类自身免疫性疾病中的抗原呈递细胞

• 批准号: 7433004
• 负责人: David A. Hafler
• 金额: $ 28.54万
• 依托单位: DANA-FARBER CANCER INST
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-09-15 至 2013-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7433004
• 关键词: Adult; Affinity; Affinity Chromatography; Alleles; Antibodies; Antigen Presentation; Antigen Targeting; Antigen-Presenting Cells; Antigens; Autoantibodies; Autoantigens; Autoimmune Diabetes; Autoimmune Diseases; Autoimmune Process; Autoimmunity; Automobile Driving; B-Lymphocytes; Binding; Biochemical; Birth; Cell Shape; Cell surface; Cells; Cerebrospinal Fluid; Child; Childhood; Clinical Trials; Clonal Expansion; Clone Cells; Collaborations; Data; Demyelinations; Dendritic Cells; Devices; Diabetes Mellitus; Disease; Epitopes; Experimental Autoimmune Encephalomyelitis; Functional disorder; Grant; Histocompatibility Antigens Class II; Human; Immunoglobulin G; Immunoglobulins; Individual; Inflammation; Inflammatory; Infusion procedures; Injury; Insulin; Insulin Antibodies; Insulin-Dependent Diabetes Mellitus; Investigation; Laboratories; Lasers; Link; Lymph Node Tissue; Measures; Methods; Microdissection; Modeling; Molecular; Multiple Sclerosis; Mus; Myelin; Nature; Outcome; Pancreas; Pathogenesis; Pathology; Pathway interactions; Patients; Peptides; Play; Population; Predisposition; Process; Production; Recombinant Antibody; Recombinants; Research Personnel; Rheumatoid Arthritis; Role; Sampling; Site; Specificity; Spleen; T-Lymphocyte; T-Lymphocyte Epitopes; Techniques; Technology; Tissues; Transgenic Mice; Work; autoreactive B cell; autoreactive T cell; base; brain tissue; clinically relevant; cohort; cytokine; diabetic; interest; interleukin-22; islet; lymph nodes; novel; novel strategies; response; tandem mass spectrometry; type I diabetic

The presence of clonally expanded populations of T cells and B cells in the inflamed tissue of patients with type 1 diabetes (diabetes) and in multiple sclerosis (MS) has long suggested that self-antigens are driving the disease process. This PPG had brought together investigators applying highly sensitive, novel techniques including the use of self-antigen tetramers and single cell cloning methods to interrogate the antibodies and T cells in the pancreatic lymph nodes of patients with diabetes and brain tissue of patients with MS. We have made significant progress in providing new evidence for the role of two candidate antigens, insulin and MOG, in these disease processes. These findings, along with the recent surprising results that infusions of anti-CD20 mAb therapy targeting B cells may have efficacy in human autoimmune diseases has led us to the underlying hypothesis of this PPG effort that alterations in the function of autoantigen-specific B cells, acting as antigen presenting cells, are central in the induction of human autoimmune diseases. In Aim 1, we will complete our initial efforts to identify the autoantigens driving clonal expansion of B cells in MS plaques and now examine diabetes islet tissue and lymph nodes. We will determine BCR sequences of individual B cells isolated by laser microdissection, followed by affinity purification of antigens with reconstructed antibodies and molecular definition of targets by tandem mass spectrometry. In Aim 2, we will interrogate the autoreactive B cells by a novel, nanowell-based technology and determine whether the B cells of patients with autoimmune diseases are pro-inflammatory. This interrogation of defined antigen specific cells will focus on functional cytokine secretion and phenotypic cell surface markers. In Aim 3, we will examine whether B cells and autoantibodies present at the sites of inflammation directly contribute to disease pathology through autoantigen presentation. In collaboration with the Kuchroo lab, we will study the ability of human recombinant antibodies generated in Aim 1 to induce demyelination in EAE models as well as the ability of B cells that express human BCRs to induce differentiation of autoreactive T cells into pathogenic effectors. High-affinity insulin-specific autoantibodies recognize the same insulin A1-15 chain epitope as human T cells, and we will examine whether these BCRs protect the T cell epitope from premature degradation during antigen presentation, a question that is clinically relevant because the affinity of insulin autoantibodies correlates with progression to diabetes. The clear association of autoantibodies with predicting susceptibility to autoimmune diabetes and the presence of anti- MOG autoantibodies in MS plaque tissue together suggest the importance of investigating B cells in these human autoimmune diseases.

患者发炎组织中存在克隆扩增的 T 细胞和 B 细胞群 长期以来，1 型糖尿病（糖尿病）和多发性硬化症 (MS) 一直表明自身抗原是驱动因素 疾病过程。该 PPG 汇集了研究人员，应用高度敏感、新颖的 技术包括使用自身抗原四聚体和单细胞克隆方法来询问 糖尿病患者胰腺淋巴结和患者脑组织中的抗体和T细胞 与女士。我们在为两名候选人的作用提供新证据方面取得了重大进展 抗原、胰岛素和 MOG 参与这些疾病过程。这些发现，以及最近令人惊讶的 结果表明，输注针对 B 细胞的抗 CD20 mAb 疗法可能对人类自身免疫性疾病有效 疾病使我们得出了 PPG 工作的基本假设： 自身抗原特异性 B 细胞作为抗原呈递细胞，在诱导人类 自身免疫性疾病。在目标 1 中，我们将完成初步工作，以确定驱动克隆的自身抗原 扩增 MS 斑块中的 B 细胞，现在检查糖尿病胰岛组织和淋巴结。我们将 确定通过激光显微切割分离的单个 B 细胞的 BCR 序列，然后进行亲和力分析 使用重建抗体纯化抗原并通过串联质量对靶标进行分子定义 光谱测定法。在目标 2 中，我们将通过一种新颖的、基于纳米孔的技术来检测自身反应性 B 细胞 并确定自身免疫性疾病患者的 B 细胞是否具有促炎性。这 对确定的抗原特异性细胞的询问将集中于功能性细胞因子分泌和表型细胞 表面标记。在目标 3 中，我们将检查 B 细胞和自身抗体是否存在于 炎症通过自身抗原的呈现直接导致疾病病理。与合作 在 Kuchroo 实验室，我们将研究 Aim 1 中产生的人类重组抗体诱导 EAE 模型中的脱髓鞘以及表达人类 BCR 的 B 细胞诱导脱髓鞘的能力 自身反应性 T 细胞分化为致病效应细胞。高亲和力胰岛素特异性自身抗体 识别与人类 T 细胞相同的胰岛素 A1-15 链表位，我们将检查这些 BCR 是否 保护 T 细胞表位在抗原呈递过程中免于过早降解，这是临床上的一个问题 相关，因为胰岛素自身抗体的亲和力与糖尿病的进展相关。清晰的 自身抗体与预测自身免疫性糖尿病易感性以及抗-抗体的存在之间的关系 MS 斑块组织中的 MOG 自身抗体表明研究这些疾病中 B 细胞的重要性 人类自身免疫性疾病。