B Cell Trafficking to the Eye

B 细胞贩运至眼睛

• 批准号: 8309047
• 负责人: JUSTINE R SMITH
• 金额: $ 1.41万
• 依托单位: OREGON HEALTH & SCIENCE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-09-01 至 2013-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8309047
• 关键词: Activated-Leukocyte Cell Adhesion Molecule; Adverse effects; Affect; Antibodies; Attention; Autoimmune Diseases; Autoimmune Process; B-Lymphocyte Subsets; B-Lymphocytes; Binding; Biologic Development; Biological Assay; Blindness; Blocking Antibodies; Blood; Blood Circulation; CD4 Positive T Lymphocytes; CXCL13 gene; Cell Adhesion Molecules; Cell Communication; Cells; Chimeric Proteins; Choroid; Clinical; Communicable Diseases; Complement; Development; Disease; Disease remission; Endothelial Cells; Endothelium; Enzyme-Linked Immunosorbent Assay; Event; Experimental Animal Model; Exploratory/Developmental Grant; Extravasation; Eye; Eye diseases; Fundus; Genes; Histopathology; Human; Image; Immunization; Immunoglobulins; Immunohistochemistry; Immunosuppressive Agents; In Vitro; Individual; Inflammation; Inflammatory; Inflammatory Infiltrate; Injection of therapeutic agent; Integrins; Intercellular adhesion molecule 1; Investigation; Laboratories; Lead; Lentivirus Vector; Leucocytic infiltrate; Leukocytes; Ligand Binding; Lymphoid; Malignant Neoplasms; Methods; Migration Assay; Modeling; Molecular; Monitor; Movement; Mus; Organ; Outcome; Pathogenesis; Patients; Pharmaceutical Preparations; Pharmacotherapy; Plasma Cells; Play; Posterior Uveitis; Posterior eyeball segment structure; Process; Proteomics; Protocols documentation; Publishing; Regulation; Research; Retina; Retinal; Risk; Role; Selectins; Shotguns; Site; Stream; Subfamily lentivirinae; System; T-Lymphocyte; Testing; Therapeutic; Time; Tissues; Toxic effect; Uveitis; Vascular Cell Adhesion Molecule-1; Vascular Endothelium; Vision; Work; base; cell motility; chemokine; clinically relevant; disease mechanisms study; disorder control; granulocyte; improved; in vivo; interest; junctional adhesion molecule; macrophage; microbial; migration; monocyte; novel; research study; trafficking; transgene expression; unpublished works; uveoretinitis

PROJECT SUMMARY Autoimmune posterior uveitis is an inflammatory eye disease affecting the posterior eye that is difficult to treat and commonly results in vision loss. The overall objective of our work is to improve the clinical outcome for patients with this condition. Posterior uveitis is characterized by a heterogeneous leukocytic infiltrate within the retina and adjacent tissues. Previous research relating to disease pathogenesis has centered on CD4+ T cells and monocytes. Mechanisms involving B cells have received little attention, although new treatments directed against these cells are therapeutic for patients with uveitis. This exploratory project will initiate a new line of research on the role of the B cell in autoimmune posterior uveitis, with initial focus on B cell migration to the retina. The central hypothesis of the proposal is that B cell entry into the retina in autoimmune posterior uveitis is strictly regulated by distinct adhesion molecules on retinal vascular endothelium. A novel murine model of B cell trafficking to the eye is proposed, in which the potent B cell chemokine, CXCL13, will be delivered to the posterior eye by intraocular injection of chemokine-encoding lentivirus. Multiple conditions will be tested to optimize the immunization protocol for maximum influx of B cells. Transgene expression will be monitored by ELISA and immunohistochemistry, and inflammation will be monitored by topical endoscopic fundus imaging and histopathology. Parallel studies of B cell-retinal endothelial interactions will be conducted using human systems, including: a retinal endothelial cell transmigration assay with primary human endothelial cells; and a modified Woodruff-Stamper assay using intact human retina. The role of selected adhesion molecules - intercellular adhesion molecule (ICAM)-1, vascular cell adhesion molecule (VCAM)-1 and activated leukocyte cell adhesion molecule (ALCAM) - in B cell trafficking to retina will be evaluated in the murine model and the human-based assays, using appropriate methods to specifically inhibit adhesion molecule-ligand binding.

项目概要 自身免疫性后葡萄膜炎是一种影响后眼的炎症性眼病，难以治疗 并通常导致视力丧失。我们工作的总体目标是改善临床结果 有这种情况的患者。后葡萄膜炎的特点是葡萄膜内异质性白细胞浸润。 视网膜和邻近组织。先前与疾病发病机制相关的研究主要集中在 CD4+ T 细胞 和单核细胞。尽管新的治疗方法针对 B 细胞，但涉及 B 细胞的机制却很少受到关注。 对抗这些细胞对葡萄膜炎患者有治疗作用。这个探索性项目将启动一条新的生产线 研究 B 细胞在自身免疫性后葡萄膜炎中的作用，最初关注 B 细胞迁移至 视网膜。该提案的中心假设是 B 细胞在自身免疫性后葡萄膜炎中进入视网膜 受到视网膜血管内皮上不同粘附分子的严格调节。 B的新型小鼠模型 提议将细胞运输到眼睛，其中有效的 B 细胞趋化因子 CXCL13 将被输送到眼睛 通过眼内注射趋化因子编码的慢病毒进行后眼观察。将测试多种条件 优化免疫方案以获得最大 B 细胞流入。转基因表达将通过以下方式监测 ELISA 和免疫组织化学，以及炎症将通过局部内窥镜眼底成像进行监测 和组织病理学。 B 细胞-视网膜内皮相互作用的平行研究将使用人类进行 系统，包括：用原代人内皮细胞进行视网膜内皮细胞迁移测定；和一个 使用完整的人视网膜进行改良的 Woodruff-Stamper 测定。选定粘附分子的作用 - 细胞间粘附分子（ICAM）-1、血管细胞粘附分子（VCAM）-1和活化白细胞 细胞粘附分子 (ALCAM) - 将在小鼠模型中评估 B 细胞向视网膜的运输 基于人类的测定，使用适当的方法特异性抑制粘附分子-配体结合。