T and B cell derived GM-CSF in EAE

摘要 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 作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是发生和发展所必需的 与慢性EAE和MS患者相比，表达GM-CSF的B和T细胞数量增加 与健康的受试者。B细胞和Th细胞都被认为是EAE/MS中GM-CSF的相关来源，但 目前尚不清楚每种细胞产生的GM-CSF在多大程度上与疾病有关 病理学。为了解决这个问题，我们开发了GM-CSF条件基因敲除小鼠，这些小鼠已经用牙线 GM-CSF基因(Csf2flx)，使细胞特异性地消融GM-CSF。Csf2flx小鼠和那些 在特定细胞类型中表达Cre重组酶将导致后代在这些细胞中缺乏GM-CSF。我们是 Csf2flx小鼠与Cre驱动系的Th1、Th17和B淋巴细胞杂交，以评价GM- 从EAE中的这些细胞类型衍生的CSF，从而解决了关于细胞来源的知识差距 GM-CSF在自身免疫性神经炎中的作用。 我们提出了以下具体目标： 具体目的1.比较Th1和Th17细胞在EAE中作为GM-CSF来源的相关性。 尽管大多数Th血统都能产生GM-CSF，但人们认为它在EAE中的来源是Th1和Th17 细胞。我们推测Th17细胞是EAE中GM-CSF的相关细胞来源，而GM-CSF Th1细胞的产生是必不可少的。为了测试这一点，到目前为止，我们已经在Tbet中产生了缺乏GM-CSF的小鼠- 表达细胞(Th1细胞)。我们目前正在完成缺乏IL-17A-GM-CSF的小鼠的发育 表达细胞(Th17细胞)。这些GM-CSF条件性基因敲除株将用于比较相关性 Th1和Th17细胞作为EAE患者GM-CSF的来源。 具体目的2.阐明B细胞来源的GM-CSF在EAE中的作用。B细胞在多发性硬化症中的重要作用 和EAE，但其在这些疾病中的致病机制尚不完全清楚。老鼠和人 B细胞表达GM-CSF，MS患者产生GM-CSF的B细胞数量增加。一直以来 提出来自B细胞的GM-CSF在EAE/MS中起致病作用，但这一假说尚未得到验证， EAE中B细胞产生GM-CSF的特征也尚未确定。我们的初步数据显示，B细胞，在 外周和中枢神经系统在EAE过程中产生GM-CSF。我们假设B细胞中的GM-CSF对 对EAE的发病具有重要意义。为了验证这一假设，我们目前培育了可诱导删除GM基因的小鼠。 B细胞中的脑脊液。

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.