REGULATION OF MACROPHAGE FUNCTION BY PROTEIN KINASES

宏观经济可以被刺激到一种高度的状态， 杀微生物和杀肿瘤活性， 定义为激活。 细胞内事件， 巨噬细胞活化的调节尚未确定。 我们建议系统地定义快速变化， 蛋白质磷酸化与刺激 巨噬细胞，并确定蛋白激酶参与这些 流程. 在这些研究中将使用两种不同的刺激; 集落刺激因子（CSF-1）和细菌脂多糖 （LPS）。 LPS和CSF-1诱导许多常见的功能变化， 巨噬细胞，但每一个也诱导独特的功能。 为 例如，只有LPS诱导杀肿瘤活性。 为了 确定CSF-1和LPS如何改变巨噬细胞活性， 将采用药理学和生物化学方法。 的 药理学方法将利用可以模拟或 抑制LPS或CSF-1的活性，以帮助识别第二种 信使/蛋白激酶系统可能介导的行动 CSF-1和LPS。 生物化学方法将决定 蛋白质磷酸化的快速变化， 刺激。 经过初步的药理学和生物化学研究， 给出了可能的蛋白激酶介导特异性 行动，蛋白激酶参与将被确定。 这将 通过比较体内磷酸化模式 通过刺激特异性的 蛋白激酶在体外细胞制剂中的作用。 蛋白激酶 将被刺激的将被选择基于结果的 药理学和磷酸化研究。 比较将 在体外和体内磷酸化之间进行， 确定磷酸化是否在同一磷蛋白上 和磷蛋白的同一个位点。 这些结果将 使我们能够确定磷酸化反应是否是由 所研究的特定蛋白激酶。 希望这将 帮助定义巨噬细胞功能被 监管.

Macrophages can be stimulated to states of heightened microbicidal and tumoricidal activity that are functionally defined as activated. The intracellular events by which macrophage activation is regulated have not yet been defined. We propose to systematically define the rapid alterations in protein phosphorylation that are associated with stimulation of macrophages and identify the protein kinases involved in these processes. Two different stimuli will be used in these studies; colony stimulating factor (CSF-1) and bacterial lipopolysaccharide (LPS). LPS and CSF-1 induce many common functional changes in macrophages, but each also induces unique functions. For example, only LPS induces tumoricidal activity. In order to determine how CSF-1 and LPS alter macrophage activity both a pharmacologic and biochemical approach will be employed. The pharmacologic approach will utilize agents that may mimic or inhibit activities of LPS or CSF-1 to help identify the second messenger/protein kinase systems that might mediate the actions of CSF-1 and LPS. The biochemical approach will determine the rapid alterations in protein phosphorylation induced by each stimuli. After initial pharmacologic and biochemical studies have given an indication of possible protein kinases mediating specific actions, the protein kinases involved will be identified. This will be accomplished by comparing the in vivo phosphorylation pattern with the phosphorylation pattern obtained by stimulating specific protein kinases in cell preparations in vitro. The protein kinases that will be stimulated will be chosen based on the results of the pharmacologic and phosphorylation studies. Comparisons will be made between the in vitro and in vivo phosphorylations to determine if the phosphorylations are on the same phosphoprotein and on the same site of that phosphoprotein. These results will allow us to determine if the phosphorylation reaction is mediated by the specific protein kinase studied. It is hoped that this will help define the mechanism(s) by which macrophage function is regulated.