Regulation of Migration of Autoreactive Germinal Centers and Precursor B Cells

自身反应性生发中心和前体 B 细胞迁移的调节

• 批准号: 8195548
• 负责人: John D Mountz
• 金额: --
• 依托单位: BIRMINGHAM VA MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-10-01 至 2013-09-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8195548
• 关键词: Address; Affinity; Antigen Presentation; Antigens; Area; Autoantibodies; Autoantigens; Autoimmune Diseases; Autoimmune Process; Autoimmunity; B-Lymphocyte Subsets; B-Lymphocytes; Behavior; CD4 Positive T Lymphocytes; CXCL12 gene; CXCL13 gene; Cell Maturation; Cell physiology; Cells; Chemotaxis; Collection; Complement 3d Receptors; Confocal Microscopy; Data; Dendritic Cells; Development; Event; Exhibits; GTP-Binding Protein Regulators; Generations; Goals; Homing; Human; Immunoglobulin M; In Vitro; Interferon-alpha; Interferons; Interleukin-17; Letters; Lupus; Lymphocyte; Maintenance; Mediating; Mediator of activation protein; Modeling; Mus; National Institute of Allergy and Infectious Disease; Patients; Phenotype; Play; Population; Prevention; Production; Publications; RGS Family Gene; Regulation; Research Personnel; Rheumatoid Arthritis; Role; Sampling; Series; Signal Transduction; Sinus; Spleen; Structure of germinal center of lymph node; Systemic Lupus Erythematosus; T-Cell Proliferation; TNFRSF5 gene; Techniques; Testing; Therapeutic Intervention; Time; Transportation; Up-Regulation; abstracting; clinically relevant; in vivo; migration; model development; mouse model; novel; novel therapeutic intervention; peripheral blood; public health relevance; response; therapeutic target

Abstract. The overall goal of this proposal is to identify the mechanisms by which IFN¿ and IL-17 act in a coordinated fashion to initiate the development of autoantibody (autoAb)-forming B cells in both human and mouse that develop systemic lupus erythematosus (lupus or SLE). Both IL-17R and IFN¿R signaling have been implicated in the formation of spontaneous germinal centers (GCs) in the BXD2 mouse model of lupus through analysis of IFN¿R-deficient and IL-17R-deficient BXD2 mice. There are high numbers of plasmacytoid DCs in the marginal sinus of the BXD2 mice that produce locally high levels of IFN¿. IFN¿ appears to influence the formation of GCs by promoting the migration of a population of CD21hiIgMhiCD23hi B cells from the marginal zone (MZ) to the follicle (FO). These B cells were referred to as CD23hiAPChi B cells as they have a very high capacity for antigen (Ag) presentation and the ability to induce a much stronger TH-17 polarization response than the conventional FO B cells. Parallel analyses indicate that IL-17 promotes the formation and stabilization of the GCs in the BXD2 mice through upregulation of regulators of G-protein signaling (Rgs)13 and Rgs16 in the B cells, which then promote the retention of APChi B cells in the FO area. We hypothesize that IFNa facilitates the release of the CD23hiAPChi B cells from the MZ and that IL-17 then localizes these cells in the FO where the CD4 T cells reside. As these CD23hiAPChi B cells have increased APC and costimulatory functions, they promote the generation/maintenance of TH-17 cells and the formation of spontaneous GCs. Three major questions will be addressed: (1) Does the IFN¿-induced influx of the CD23hiAPChi B cells initiate the development of GCs in BXD2 mice? (2) Do the CD23hiAPChi B cells respond to IL-17 and promote polarization of TH-17 cells leading to the production of autoAbs in the BXD2 mice? And (3) Can IFNa and IL-17 regulate the migration behavior of APChi B cells in lupus patients? We will analyze Ag capture, transportation and presentation by APChi B cells to CD4 T cells in vivo using confocal microscopy and FACS analysis. The ability of IL-17 and IFNa to regulate chemotaxis will be determined in vitro, and confirmed in vivo by analysis of homing of GFP-positive APChi B cells using B cells from BXD2-Il-17r-/-, BXD2-Ifnar-/-, BXD2-Rgs13-/- and BXD2-Rgs16-/- mice. The clinical relevance of this model of the development of autoimmunity will be assessed by that analysis of B cells obtained from the peripheral blood of lupus patients for the analysis of the effects of IFN¿ and IL-17 on the development and migration of APChi B cells. SIGNIFICANCE: These studies should identify critical migratory signals and upstream mediators that facilitate the development of an autoimmune circuit that leads to the production of high affinity pathogenic autoAbs. Thus, they have the potential to identify novel candidate therapeutic targets and to suggest more effective therapeutic interventions in autoAb-mediated autoimmune diseases.

摘要：该提案的总体目标是确定 IFN 的作用机制。和 IL-17 的作用 协调方式启动人类自身抗体 (autoAb) 形成 B 细胞的发育 以及患有系统性红斑狼疮（狼疮或 SLE）的小鼠 IL-17R 和 IFN¿ R信号传导 与 BXD2 小鼠模型中自发生发中心 (GC) 的形成有关 通过 IFN 分析诊断狼疮R 缺陷和 IL-17R 缺陷的 BXD2 小鼠数量较多。 BXD2 小鼠边缘窦中的浆细胞样 DC 产生局部高水平的 IFN¿ . 干扰素? 似乎通过促进群体的迁移来影响GC的形成 CD21hiIgMhiCD23hi B 细胞从边缘区 (MZ) 到滤泡 (FO) 这些 B 细胞被称为。 作为 CD23hiAPChi B 细胞，因为它们具有非常高的抗原 (Ag) 呈递能力以及 诱导比传统 FO B 细胞更强的 TH-17 极化反应。 表明 IL-17 通过促进 BXD2 小鼠 GC 的形成和稳定 B 细胞中 G 蛋白信号传导 (Rgs)13 和 Rgs16 的调节因子上调，然后促进 APChi B 细胞保留在 FO 区域，我们勇敢地说 IFNa 促进了 IFNa 的释放。 来自 MZ 的 CD23hiAPChi B 细胞和 IL-17 然后将这些细胞定位在 FO 中，其中 CD4 由于这些 CD23hiAPChi B 细胞具有增强的 APC 和共刺激功能， 它们促进 TH-17 细胞的生成/维持以及自发 GC 的形成。 将解决三个主要问题：(1) IFN 是否有效？ -诱导CD23hiAPChi B细胞的流入 (2) CD23hiAPChi B 细胞对 IL-17 和 IL-17 是否有反应？ 促进 TH-17 细胞极化，从而导致 BXD2 小鼠产生自身抗体？ (3) 可以吗？ IFNa和IL-17调节狼疮患者APChi B细胞的迁移行为？我们将分析Ag？ 使用共聚焦显微镜在体内捕获、运输 APChi B 细胞并将其呈递给 CD4 T 细胞 和FACS分析将在体外测定IL-17和IFNa调节趋化性的能力。 通过使用来自 BXD2-Il-17r-/- 的 B 细胞分析 GFP 阳性 APChi B 细胞的归巢在体内证实， BXD2-Ifnar-/-、BXD2-Rgs13-/- 和 BXD2-Rgs16-/- 小鼠模型的临床相关性。 自身免疫的发展将通过对从外周获得的 B 细胞进行分析来评估 狼疮患者的血液用于分析 IFN 的效果¿和 IL-17 的发育和迁移 APChi B 细胞的意义：这些研究应该确定关键的迁移信号和上游信号。 促进自身免疫回路发展的介质，从而产生高 因此，它们有潜力识别新的候选治疗靶点。 并提出针对自身抗体介导的自身免疫性疾病更有效的治疗干预措施。