MOLECULAR PATHOLOGY OF LPS INDUCED INJURY

LPS 引起的损伤的分子病理学

• 批准号: 2060170
• 负责人: Richard J Ulevitch
• 金额: $ 41.24万
• 依托单位: SCRIPPS RESEARCH INSTITUTE, THE
• 依托单位国家: 美国
• 财政年份: 1978
• 资助国家: 美国
• 起止时间: 1978-08-01 至 1995-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2060170
• 关键词: CD antigens; bacterial disease; bacterial polysaccharides; binding proteins; chemical binding; circular dichroism; disease /disorder model; disseminated intravascular coagulation; endotoxins; fluorescence spectrometry; gene expression; genetic manipulation; gram negative bacteria; laboratory mouse; laboratory rabbit; laboratory rat; lipopolysaccharides; macrophage; molecular cloning; monoclonal antibody; protein sequence; tissue /cell culture; tumor necrosis factor alpha

The long term goal for our research is to understand bacterial lipopolysaccharide (LPS) action at the molecular level. We will focus our efforts on models that should help us understand how LPS induces cellular injury during endotoxemia resulting from gram-negative bacillary infections. We hypothesize that a significant contribution to the initial host response to LPS is described by three sequential events; FIRST, the formation of a complex between LPS and a plasma protein we discovered, characterized and named LPS-binding protein (LBP). SECOND, the binding of LPS-LBP complexes to monocytes/macrophages via a unique plasma membrane receptor we have identified as CD14. THIRD, stimulation of macrophages by LPS to rapidly release mediators such as TNF. To test this hypothesis we will prepare monoclonal antibodies to LBP and CD14 for initial use in two types of in vitro experiments; (i). to investigate their ability to block binding of LPS to LBP and of LPS-LBP to CD14 on macrophages, (ii). to identify the structures of LBP involved in LPS binding and binding of LPS-LBP complexes to CD14. This latter approach will provide the requisite data for design of polypeptides to be tested as inhibitors of LPS/LBP/CD14 interactions. To further test our hypothesis we will use a rabbit model of endotoxic shock that approximates pathophysiologic changes in man during endotoxemia to test whether antibodies and/or polypeptides shown to inhibit LPS/LBP/CD14 interactions in vitro will modify LPS-induced release of TNF, DIC and lethality. Implicit in these studies is the potential for development of new therapeutic modalities to intervene in endotoxemia in man.

我们研究的长期目标是了解细菌 脂多糖（LPS）在分子水平上作用。我们将集中精力 对应该有助于我们了解LP诱导细胞的模型的努力 革兰氏阴性杆菌引起的内毒素血症期间的损伤 感染。我们假设对初始的重要贡献 主机对LPS的反应由三个顺序事件描述。首先， 我们发现的LPS和血浆蛋白之间的复合物的形成， 表征并命名为LPS结合蛋白（LBP）。第二，结合 通过独特的质膜到单核细胞/巨噬细胞到单核细胞/巨噬细胞的LPS-LBP复合物 受体我们已确定为CD14。第三，通过 LPS迅速释放诸如TNF之类的介体。为了检验这一假设 将准备对LBP和CD14的单克隆抗体，以初次使用 体外实验的类型； （我）。调查他们阻止的能力 LPS与LBP和LPS-LBP与巨噬细胞的CD14结合，（ii）。到 确定参与LPS结合和结合的LBP结构 LPS-LBP复合物至CD14。后一种方法将提供必要的 用于设计为LPS/LBP/CD14抑制剂的多肽设计的数据 互动。为了进一步检验我们的假设，我们将使用 内毒性休克近似于人类的病理生理变化 内毒素血症测试抗体和/或多肽是否抑制 LPS/LBP/CD14体外相互作用将改变LPS诱导的TNF的释放， dic和杀伤力。这些研究中隐含的潜力 开发新的治疗方式以干预内毒素血症 男人。