T and B cell derived GM-CSF in EAE

总结 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， 作为自身免疫性疾病的主要致病因素，包括多发性硬化（MS）及其动物模型， 模型实验性自身免疫性脑脊髓炎（EAE）。GM-CSF是发展所必需的， EAE的慢性化和MS患者相比，表达GM-CSF的B和T细胞数量增加， 健康的受试者。B和Th细胞都被认为是EAE/MS中GM-CSF的相关来源，但 目前尚不清楚这些细胞类型中的每一种产生GM-CSF在多大程度上有助于疾病， 病理为了解决这个问题，我们开发了GM-CSF条件性敲除小鼠， GM-CSF基因（Csf 2flox），其能够实现GM-CSF的细胞特异性消融。Csf 2flox小鼠和那些 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后代。我们 将Csf 2 flox小鼠与Th 1、Th 17和B淋巴细胞的Cre驱动细胞系杂交，以评估GM-1的作用。 EAE中源自这些细胞类型的CSF，从而解决了关于细胞来源的知识空白 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Th 1和Th 17细胞作为GM-CSF来源与EAE的相关性。 尽管大多数Th细胞系都能产生GM-CSF，但据信其在EAE中的来源是Th 1和Th 17 细胞我们推测，Th 17细胞是EAE中GM-CSF的相关细胞来源，而GM-CSF在EAE中的表达与Th 17细胞的表达有关。 Th 1细胞的产生是不稳定的。为了测试这一点，我们迄今为止已经产生了Tbet中缺乏GM-CSF的小鼠。 表达细胞（Th 1细胞）。我们目前正在完成缺乏GM-CSF的小鼠的IL-17 A-1的开发。 表达细胞（Th 17细胞）。这些GM-CSF条件性敲除细胞系将用于比较 Th 1和Th 17细胞是EAE中GM-CSF的来源。 具体目标2。目的：探讨B细胞GM-CSF在EAE中的作用。B细胞在MS中起重要作用 和EAE，但它们在这些疾病中的致病机制尚不完全清楚。小鼠和人 B细胞表达GM-CSF，并且MS患者具有增加数量的产生GM-CSF的B细胞。已经 提出来自B细胞的GM-CSF在EAE/MS中起致病作用，但这一假设尚未得到验证， EAE中B细胞的GM-CSF产生也没有特征。我们的初步数据显示，在正常人中，B细胞 外周和CNS在EAE期间产生GM-CSF。我们假设来自B细胞的GM-CSF 对EAE发病有重要意义。为了验证这一假设，我们目前培育了诱导性缺失GM的小鼠， B细胞中的CSF。

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.