T and B cell derived GM-CSF in EAE

EAE 中 T 细胞和 B 细胞衍生的 GM-CSF

• 批准号: 9809336
• 负责人: Bogoljub Ciric
• 金额: $ 23.4万
• 依托单位: THOMAS JEFFERSON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-06-20 至 2021-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9809336
• 关键词: Ablation; Address; Adjuvant; Animal Model; Attenuated; Autoimmune Diseases; Autoimmune Process; Autoimmunity; B-Lymphocytes; Biological; Blood; Bone Marrow; Cell physiology; Cells; Chronic; Clinical; Clinical Trials; Colony-Stimulating Factor Gene; Colony-Stimulating Factor Receptors; Data; Development; Disease; Effector Cell; Enterobacteria phage P1 Cre recombinase; Experimental Animal Model; Experimental Autoimmune Encephalomyelitis; Genes; Granulocyte-Macrophage Colony-Stimulating Factor; Human; In Vitro; Individual; Infection; Inflammation; Inflammatory; Interleukin-17; Interleukin-4; Knock-out; Knockout Mice; Knowledge; Lead; LoxP-flanked allele; Multiple Sclerosis; Mus; Myelogenous; Myeloid Cells; Onset of illness; Pathogenesis; Pathogenicity; Pathology; Phase; Phase II Clinical Trials; Play; Population; Process; Production; Proteins; Rheumatoid Arthritis; Role; Safety; Source; Stimulus; Symptoms; T-Lymphocyte; T-Lymphocyte Subsets; Testing; Th1 Cells; Tissues; Virulence Factors; Work; cell type; cytokine; efficacy testing; in vivo; knockout animal; monocyte; mouse development; multiple sclerosis patient; neuroinflammation; receptor; response; therapeutic target; therapy outcome; trend; virtual

SUMMARY Granulocyte-macrophage colony-stimulating factor (GM-CSF, CSF-2) is a pro-inflammatory cytokine that has emerged as the major pathogenic factor in autoimmune diseases, including multiple sclerosis (MS) and its animal model experimental autoimmune encephalomyelitis (EAE). GM-CSF is required for the development and chronicity of EAE, and MS patients have increased numbers of GM-CSF-expressing B and T cells compared with healthy subjects. Both B and Th cells have been proposed as relevant sources of GM-CSF in EAE/MS, but it is currently unknown to what degree GM-CSF production by each of these cell types contributes to disease pathology. To address this question, we have developed GM-CSF conditional knockout mice, which have floxed GM-CSF gene (Csf2flox) that enable cell-specific ablation of GM-CSF. A cross between Csf2flox mice and those that express Cre recombinase in specific cell types will result in progeny lacking GM-CSF in those cells. We are crossing Csf2flox mice with Cre driver lines for Th1, Th17 and B lymphocytes in order to evaluate the role of GM- CSF derived from these cell types in EAE, thereby addressing a knowledge gap regarding the cellular sources of GM-CSF in autoimmune neuroinflammation. We propose the following specific aims: Specific Aim 1. To compare the relevance of Th1 and Th17 cells as the sources of GM-CSF in EAE. Although GM-CSF can be produced by most Th lineages, it is believed that its sources in EAE are Th1 and Th17 cells. We hypothesize that Th17 cells are the relevant cellular source of GM-CSF in EAE, while GM-CSF production by Th1 cells is dispensable. To test this, we have thus far generated mice lacking GM-CSF in Tbet- expressing cells (Th1 cells). We are currently finishing development of mice lacking GM-CSF in IL-17A- expressing cells (Th17 cells). These GM-CSF conditional knockout lines will be used to compare the relevance of Th1 and Th17 cells as the sources of GM-CSF in EAE. Specific Aim 2. To elucidate the role of GM-CSF from B cells in EAE. B cells play an important role in MS and in EAE, but their pathogenic mechanisms in these diseases are incompletely understood. Mouse and human B cells express GM-CSF, and MS patients have increased numbers of GM-CSF-producing B cells. It has been proposed that GM-CSF from B cells plays a pathogenic role in EAE/MS, but this hypothesis has not been tested, nor has GM-CSF production by B cells in EAE been characterized. Our preliminary data show that B cells, in the periphery and CNS, produce GM-CSF during EAE. We hypothesize that GM-CSF from B cells contributes significantly to EAE pathogenesis. To test this hypothesis, we currently generate mice that inducibly delete GM- CSF in B cells.

概括 粒细胞巨噬细胞集落刺激因子（GM-CSF、CSF-2）是一种促炎细胞因子，具有 已成为自身免疫性疾病的主要致病因素，包括多发性硬化症（MS）及其动物 实验性自身免疫性脑脊髓炎（EAE）模型。 GM-CSF 是发育所必需的 与 EAE 慢性病和 MS 患者相比，表达 GM-CSF 的 B 细胞和 T 细胞数量增加 与健康受试者。 B 细胞和 Th 细胞均被提议作为 EAE/MS 中 GM-CSF 的相关来源，但是 目前尚不清楚每种细胞类型产生的 GM-CSF 在何种程度上导致疾病 病理。为了解决这个问题，我们开发了 GM-CSF 条件敲除小鼠， GM-CSF 基因 (Csf2flox) 可实现细胞特异性消融 GM-CSF。 Csf2flox 小鼠与那些小鼠的杂交 在特定细胞类型中表达 Cre 重组酶将导致这​​些细胞中缺乏 GM-CSF 的后代。我们是 将 Csf2flox 小鼠与 Th1、Th17 和 B 淋巴细胞的 Cre 驱动系杂交，以评估 GM-的作用 CSF 源自 EAE 中的这些细胞类型，从而解决了有关细胞来源的知识差距 GM-CSF 在自身免疫性神经炎症中的作用。 我们提出以下具体目标： 具体目标 1. 比较 Th1 和 Th17 细胞作为 EAE 中 GM-CSF 来源的相关性。 虽然 GM-CSF 可由大多数 Th 谱系产生，但据信其在 EAE 中的来源是 Th1 和 Th17 细胞。我们假设 Th17 细胞是 EAE 中 GM-CSF 的相关细胞来源，而 GM-CSF Th1细胞的产生是可有可无的。为了测试这一点，我们迄今为止在 Tbet- 中培育了缺乏 GM-CSF 的小鼠 表达细胞（Th1 细胞）。我们目前正在完成 IL-17A 中缺乏 GM-CSF 的小鼠的开发 表达细胞（Th17 细胞）。这些 GM-CSF 条件敲除系将用于比较相关性 Th1 和 Th17 细胞作为 EAE 中 GM-CSF 的来源。 具体目标 2. 阐明 B 细胞的 GM-CSF 在 EAE 中的作用。 B细胞在多发性硬化症中发挥重要作用 和 EAE，但它们在这些疾病中的致病机制尚不完全清楚。老鼠和人类 B 细胞表达 GM-CSF，MS 患者产生 GM-CSF 的 B 细胞数量增加。它一直 提出来自 B 细胞的 GM-CSF 在 EAE/MS 中发挥致病作用，但这一假设尚未得到检验， EAE 中 B 细胞产生 GM-CSF 的情况也尚未被表征。我们的初步数据表明，B 细胞 外周和中枢神经系统，在 EAE 期间产生 GM-CSF。我们假设来自 B 细胞的 GM-CSF 有助于 对 EAE 发病机制有显着影响。为了验证这一假设，我们目前培育了可诱导删除 GM- 的小鼠。 B 细胞中的脑脊液。