IMMUNOREGULATORY EFFECTS OF ESTROGEN IN EAE

描述（由申请人提供）：多发性硬化症患者 (MS)，存在明显的性别偏见，大约是 受影响的女性和男性一样。性激素可能会导致易感性， 对MS的抗性通过影响潜在的 对中枢神经系统（CNS）抗原特异的病原性T细胞，以及 作为调节性T细胞，可能会改变疾病的进程。此前我们 报道，低剂量的17 β-雌二醇（E2）可以通过以下方式降低EAE的严重程度： 抑制活化、细胞因子和趋化因子产生，和 鼠髓鞘少突胶质细胞特异性T细胞致脑炎性 糖蛋白（MOG）、蛋白脂质蛋白（PLP）或髓鞘碱性蛋白（MBP）， 以及抑制炎性细胞向CNS的募集。的 特别重要的是，雌激素治疗大大减少了细胞内 TNF-α的产生，称为EAE中的关键炎性细胞因子， 女士E2在EAE中作用的微阵列研究进展 脾细胞基因表达证实了TNF-α的有效抑制， 进一步确定了几个先前未被怀疑的免疫相关候选者， 体内E2处理似乎强烈影响基因。这些 雌激素对致病细胞、募集细胞和调节细胞的调节作用 在EAE中可能通过与雌激素受体（ER）直接相互作用介导， 包括经典的ER-α和ER-β， (iER)，以及可能与iER不同的膜ER（mER）。这 一项提案将检验雌激素对EAE的调节涉及 受体介导的调节TNF-α和其他几种新的免疫相关 基因，从而抑制巨噬细胞的炎症作用， 树突状细胞和T细胞以及增强调节性NKT细胞活性。我们 主要目的是确定E2对EAE的抑制作用是否可以被 在缺乏调节性T和NKT细胞的自发性EAE中观察到，以及 如果E2对EAE的抑制作用通过iER-α和/或iER β介导， 或者相反，没有一个经典iER。使用新开发 ER-α和ER-β基因敲除小鼠，我们将首次将E2- 依赖性调节与一个或两个iER，或可选地与mER，如果E2 在双KO小鼠中效果持续。此外，对于每个新的E2影响基因， 暗示从微阵列分析，我们将研究细胞 来源，E2对转录和蛋白质产生的影响，以及 在体内EAE诱导的贡献。发现在EAE中重要的基因 将进一步评估来自不同水平的妇女的血细胞， E2. 这项工作将确定关键的雌激素敏感基因，包括 TNF α，促进EAE诱导和抗性。这些基因的变化 可以在雌激素治疗期间作为替代标记物进行随访， 有效剂量的雌激素通过这项研究，我们将开发出一种坚实的 使用雌激素替代疗法治疗MS的基础。

DESCRIPTION (provided by applicant): In patients with multiple sclerosis (MS), there is a distinct gender bias, with approximately twice as many affected females as males. Sex hormones may contribute to susceptibility or resistance to MS by influencing development and function of potentially pathogenic T cells specific for central nervous system (CNS) antigens, as well as regulatory T cells that might modify the course of disease. Previously, we reported that low doses of 17beta-estradiol (E2) can reduce severity of EAE by inhibiting activation, cytokine and chemokine production, and encephalitogenicity of murine T cells specific for myelin oligodendrocyte glycoprotein (MOG), proteolipid protein (PLP), or myelin basic protein (MBP), and by inhibiting I recruitment of inflammatory cells into the CNS. Of particular importance, estrogen treatment profoundly reduced intracellular production of TNF-alpha, known as a critical inflammatory cytokine in EAE and MS. Recent evaluation of E2 effects on EAE using microarray analysis of splenocyte gene expression confirmed potent inhibition of TNF-alpha, and further identified several previously unsuspected immune-associated candidate genes appearing to be strongly affected by E2 treatment in vivo. These modulatory effects of estrogen on pathogenic, recruited, and regulatory cells in EAE are likely mediated y direct interaction with estrogen receptors (ER), which include the classical ER-alpha and ER-beta that are internal f receptors (iER), and possibly membrane ERs (mER) that may be distinct from iERs. This proposal will test the hypothesis that modulation of EAE by estrogen involves receptor-mediated regulation of TNF-alpha and several other novel immunerelated genes, thereby inhibiting inflammatory effects of macrophages, dendritic cells and T cells and enhancing regulatory NKT cell activity. Our primary goals are to determine if the inhibitory effects of E2 on EAE can be observed in spontaneous EAE in the absence of regulatory T and NKT cells, and if inhibitory effects of E2 on EAE are mediated through iER-alpha and /or iERbeta, or in contrast, by neither of the classical iERs. Using newly developed ER-alpha and ER-beta knockout mice we will for the first time associate E2- dependent regulation with either or both iERs, or alternatively with mER if E2 effects persist in double KO mice. Moreover, for each novel E2-affected gene implicated from the microarray analysis, we will investigate the cellular source, effects of E2 on transcription and protein production, and contribution to EAE induction in vivo. Genes found to be important in EAE will be further evaluated in blood cells from women with different levels of E2. The work proposed will identify key estrogen-sensitive genes, including TNFalpha, contributing to EAE induction and resistance. Changes in these genes can be followed during estrogen treatment as surrogate markers to verify effective doses of estrogen. From this research, we will develop a solid basis for using estrogen replacement therapy for MS.