INHIBITION OF INFLAMMATION

活化的巨噬细胞是许多炎症性疾病的共同标志，并且在内毒素休克的结果中起重要作用。该提案使用分子和细胞方法在体内和体外研究Mer受体酪氨酸激酶对巨噬细胞的下调。本研究的目的是探讨Mer受体酪氨酸激酶抑制巨噬细胞的机制以及巨噬细胞下调对炎症的影响。这项提议的发现将对减轻与巨噬细胞相关的炎症产生广泛的影响。具体目标有四个：（1）。我们将评估Mer抑制LPS信号转导的分子机制。我们将评估用mer/kd和mer/WT构建体转染的表达CD 14的CHO细胞系中IkB α和IkB β的改变。我们还将评估Mer在LPS激活的JNK、p38和ERK信号通路中的作用。最后，我们将评估Mer是否相互作用或影响CD 14的细胞表面表达。2）。我们将确定Mer在炎症过程中如何调节巨噬细胞。我们将使用经历炎症的mer/kd和mer/WT小鼠，并评估组织学、细胞因子谱和巨噬细胞活化的严重程度。此外，我们提出了一个转基因动物的诱导系统的控制下，直接确定的后果Mer表达在炎症过程中。3）。我们将确定Mer对TNF-α诱导的细胞凋亡的调节是否位于NF-κ B上游的控制点。我们将评估Mer是否改变TRAF水平以诱导NF-κ B。我们还将评估mer/kd和mer/WT小鼠炎症部位的体内细胞凋亡。4）。我们将确定Mer活性是否与慢性炎症患者的巨噬细胞活化相关。将在从患者分离的巨噬细胞中评估Mer的蛋白质和磷酸化测定。此外，还将量化细胞因子谱和NF-κ B，以确定与炎症的相关性。

Activated macrophages are a common hallmark of many inflammatory disease and play a significant role in the outcome of endotoxic shock. This proposal studies the down-regulation of macrophages by the Mer receptor tyrosine kinase using molecular and cellular methods, both in vivo and in vitro. The purpose of this proposal is to determine mechanisms by which Mer receptor tyrosine kinase inhibit macrophages and the consequence of the macrophage down-regulation on inflammation. The finding of this proposal would have broad implications toward attenuating inflammation associated with macrophages. The Specific Aims are fourfold: 1). We will assess the molecular mechanism by which Mer inhibits LPS signal transduction. We will assess alteration in IkBalpha and IkBbeta in CD14 expressing CHO cell lines transfected with mer/kd and mer/WT constructs. We will also assess the role of Mer in JNK, p38 and ERK signaling pathways activated by LPS. Finally, we will assess whether Mer is interacting or affecting the cell surface expression of CD14. 2). We will determine how Mer modulates macrophages during inflammation. We will use mer/kd and mer/WT mice undergoing inflammation and assess severity of histology, cytokine profiles and macrophage activation. In addition, we propose a transgenic animal under the control of an inducible system to directly determine the consequence of Mer expression during inflammation. 3). We will determine whether the regulation of TNF-alpha-induced apoptosis by Mer is at a control point upstream of NF-kappaB. We will assess whether Mer alters levels of TRAFs to induce NF-kappaB. Also we will assess in vivo apoptosis in sites of inflammation mer/kd and mer/WT mice. 4). We will determine whether Mer activity can be correlated to macrophage activation from patients with chronic inflammation. Protein and phosphorylation assays of Mer will be assessed in macrophages isolated from patients. In addition, cytokine profiles and NF-kappaB will be quantified to determine correlation with inflammation.