MOLECULAR ANALYSIS OF MACROPHAGE ACTIVATION VIA LBP:LPS

通过 LBP:LPS 激活巨噬细胞的分子分析

• 批准号: 3306655
• 负责人: Sanna M Goyert
• 金额: $ 11.54万
• 依托单位: NORTH SHORE UNIVERSITY HOSPITAL
• 依托单位国家: 美国
• 财政年份: 1992
• 资助国家: 美国
• 起止时间: 1992-09-30 至 1995-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3306655
• 关键词: acute phase protein; binding proteins; cell death; crosslink; differentiation antigens; genetically modified animals; glycoproteins; human tissue; interleukin 1; interleukin 6; laboratory mouse; leukocyte activation /transformation; lipopolysaccharides; macrophage; monoclonal antibody; point mutation; tissue /cell culture; transfection; tumor necrosis factor alpha

CD14 is a myelomonocytic differentiation antigen expressed strongly on the surface of monocytes and weakly on granulocytes. CD14 has recently been shown to be a receptor for a complex consisting of lipopolysaccharide (LPS) and an acute phase serum protein called LBP (lipopolysaccharide binding protein). Binding of LBP:LPS complex to monocytes via CD14 results in strong activation of macrophages as measured by the production of tumor necrosis factor (TNF). Activation of macrophages by this pathway is thought to be the initial step in the production of TNF found in endotoxic shock and mediating the collapse of the cardiovascular-pulmonary-renal systems. The primary goals of this proposal are to determine whether soluble forms of CD14 can inhibit macrophage activation via the LBP:LPS pathway and to further define this pathway of macrophage activation at the molecular level by mapping the LBP:LPS binding site on the CD14 molecule. In addition, we will define peptides which can reduce and/or inhibit macrophage activation via this pathway. Soluble CD14 and its peptides will be analyzed both in vitro and in vivo for the ability to inhibit LPS-induced macrophage activation and/or death due to endotoxin shock. The in vivo experiments will utilize a transgenic mouse which we have recently produced which expresses human CD14 and which is more susceptible to LPS than normal mice. The long term goal of this proposal is to use CD14 peptides to reduce and/or inhibit TNF production in gram negative septic shock with the prospect of allowing time for the patient to respond to appropriate alternative therapy.

CD14 是一种骨髓单核细胞分化抗原，在 单核细胞表面，粒细胞表面弱。 CD14最近 已被证明是由以下组成的复合物的受体 脂多糖 (LPS) 和称为 LBP 的急性期血清蛋白 （脂多糖结合蛋白）。 LBP:LPS 复合物与 单核细胞通过 CD14 导致巨噬细胞强烈激活 通过肿瘤坏死因子（TNF）的产生来测量。 激活 巨噬细胞通过这条途径被认为是 在内毒素休克中发现 TNF 的产生并介导细胞崩溃 心血管-肺-肾系统。 本次活动的主要目标 建议确定可溶形式的 CD14 是否可以抑制 通过 LBP:LPS 途径激活巨噬细胞并进一步定义这一点 通过绘制巨噬细胞在分子水平上的激活途径 LBP：CD14 分子上的 LPS 结合位点。 此外，我们将定义 可以通过此减少和/或抑制巨噬细胞活化的肽 途径。 可溶性 CD14 及其肽将在体外进行分析 以及体内抑制 LPS 诱导的巨噬细胞活化的能力 和/或因内毒素休克而死亡。 体内实验将 利用我们最近生产的转基因小鼠 表达人类 CD14，比正常人更容易受到 LPS 的影响 老鼠。 该提案的长期目标是利用 CD14 肽 减少和/或抑制革兰氏阴性败血性休克中 TNF 的产生 允许患者有时间对适当的反应做出反应的前景 替代疗法。