Regulation of IDDM by Proinflammatory and Th1-cytokines

描述（由申请方提供）：胰岛素依赖型糖尿病（IDDM）的特征是T淋巴细胞浸润到胰腺的胰岛（胰岛炎）中，随后选择性破坏分泌胰岛素的β细胞，导致明显的糖尿病。因此，我们观察到了胰岛素依赖型糖尿病与AIM因子的重要联系（一种我们最初鉴定为凋亡抑制因子的分泌分子），它们是：（1）与非肥胖型糖尿病（NOD）背景回交的AIM [-/-]小鼠显示出对IDDM的完全预防;（2）AIM在疾病的非常早期阶段通过胰岛中浸润的巨噬细胞表达;（3）AIM强烈诱导巨噬细胞和树突状细胞（DC）中的TNF-α、IL-1 β、IL-6和IL-12。基于这些，出现了一种假设，即AIM可能通过在疾病发作阶段诱导胰岛中最初浸润的巨噬细胞和DC中的促炎细胞因子和I型细胞因子来加速IDDM。具体目标1和2的重点是通过在预期疾病复发的AIM无效NOD小鼠的巨噬细胞中添加回AIM表达来建立假设的适当性（目标1），以及通过评估巨噬细胞和DC中细胞因子经由泰特-诱导型转基因系统可以克服AIM-null NOD小鼠中的疾病预防（目的2）。此外，我们最近的研究结果表明，AIM介导的Toll信号转导诱导细胞因子引起的想法，推定的AIM受体可能是一个Toll/IL-1受体家族成员。在具体目标3中，我们将通过对巨噬细胞产生的cDNA文库进行表达筛选来纯化和表征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，因为我们通过使用细胞产生AIM[-/-]/-NOD。我们提出的研究将阐明在AIM背景下，特别是在疾病的早期阶段，IDDM发病机制的确切情况，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家族生理功能的一个新方面。

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.