Regulation of IDDM by Proinflammatory and Th1-cytokines

描述（由申请人提供）：胰岛素依赖型糖尿病（IDDM）的特征是t淋巴细胞浸润到胰腺朗格汉斯岛（胰岛素炎），随后选择性破坏分泌胰岛素的β细胞，导致显性糖尿病。我们初步观察到IDDM与AIM因子（一种我们最初确定为凋亡抑制因子的分泌分子）之间的重要关联，即：(1)AIM[-/-]小鼠回交至非肥胖型糖尿病（NOD）背景，显示出IDDM的完全预防；(2) AIM在疾病早期通过浸润胰岛巨噬细胞表达；(3) AIM强烈诱导巨噬细胞和树突状细胞（dc）的tnf - α、IL-1 β、IL-6和IL-12。基于这些，出现了一种假设，即AIM可能在疾病发病阶段通过诱导最初浸润的巨噬细胞和胰岛dc中的促炎和I型细胞因子加速IDDM。Specific Aim 1和2的重点是通过在期待疾病复发的Aim -null NOD小鼠中加入巨噬细胞中的Aim表达来建立假设的适当性（Aim 1），并通过评估通过tet诱导的转基因系统诱导巨噬细胞和dc中的细胞因子是否可以克服Aim -null NOD小鼠的疾病预防来测试Aim信号下游细胞因子对疾病加速的影响（Aim 2）。此外，我们最近的结果表明，AIM介导toll信号传导诱导细胞因子，这引发了一个想法，即假定的AIM受体可能是TolI/IL-1受体家族成员。在Specific Aim 3中，我们将通过巨噬细胞生成的cDNA文库的表达筛选纯化和表征Aim受体。我们还计划通过破坏nod衍生的ES细胞中的基因来创建AIM受体的敲除小鼠，因为我们利用这些细胞产生了AIM[-/-]/- nod。我们提出的研究将阐明AIM背景下IDDM发病机制的确切图景，特别是在疾病的早期阶段，因此将有助于通过抑制AIM开发新的治疗方法。此外，AIM受体的鉴定将揭示AIM功能的精确分子机制，以及toll家族生理功能的新方面。

DESCRIPTION (provided by applicant): Insulin-dependent diabetes mellitus (IDDM) is characterized by the infiltration of T-lymphocytes into the islets of Langerhans of the pancreas (insulitis), followed by selective destruction of insulin-secreting beta cells leading to overt diabetes. Preliminarily, we observed the important association of IDDM with AIM factor (a secretion molecule we initially identified as an apoptosis inhibitory factor), which are: (1) AIM [-/-] mice backcrossed to non-obese diabetes (NOD) background showed complete prevention of IDDM; (2) AIM is expressed by infiltrating macrophages in the pancreatic islets from the very early stage of the disease; (3) AIM strongly induces TNF-alpha, IL-1 beta, IL-6 and IL-12 in macrophages and dendritic cells (DCs). Based on these, a hypothesis has emerged that AIM may accelerate IDDM by inducing pro-inflammatory- and type I- cytokines in initially infiltrating macrophages and DCs in the islets at the onset stage of the disease. The Specific Aim 1 and 2 are focused on establishing the propriety of the hypothesis by adding back the AIM expression in macrophages in AIM-null NOD mice expecting the disease recurrence (aim 1), and testing the impact of the cytokines downstream of AIM signaling on the disease acceleration by assessing whether the induction of the cytokines in macrophages and DCs via the Tet-inducible transgenic system may overcome the disease prevention in AIM-null NOD mice (aim 2). In addition, our recent result that AIM mediates Toll-signaling to induce the cytokines provoked an idea that the putative AIM-receptor may be a TolI/IL-1 receptor family member. In the Specific Aim 3, we will purify and characterize the AIM-receptor by expression screening of a cDNA library generated from macrophage cells. We also plan to create knockout mice of the AIM-receptor by disrupting the gene in the NOD-derived ES cells, as we generated AIM[-/-]/-NOD by using the cells. Our proposed studies will clarify the precise picture of the IDDM pathogenesis in the context of AIM, in particular, during the early stage of the disease, and thus will contribute to development of a new therapy via suppression of AIM. In addition, identification of AIM-receptor will shed light on the precise molecular machinery of AIM functions, as well as a new aspect of physiological function of the Toll-family.