IMMUNOCHEMISTRY OF ENDOTOXIN UNRESPONSIVE C3H/HEJ MICE

内毒素无反应 C3H/HEJ 小鼠的免疫化学

• 批准号: 2003371
• 负责人: DAVID C. MORRISON
• 金额: $ 30.67万
• 依托单位: UNIVERSITY OF KANSAS MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 1986
• 资助国家: 美国
• 起止时间: 1986-01-01 至 1999-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2003371
• 关键词: B lymphocyte; antiidiotype antibody; binding proteins; crosslink; endotoxins; hamsters; immune tolerance /unresponsiveness; immunochemistry; immunotoxicity; laboratory mouse; laboratory rabbit; lipopolysaccharides; molecular cloning; monoclonal antibody; protein purification; receptor; transfection

Bacterial endotoxic lipopolysaccharides (LPS) can initiate fever, profound hypotension (shock), disseminated intravascular coagulation, multisystem organ failure and death. There is substantial evidence to support endotoxin-induced host responses as a major contributing factor to the more than 25,000 deaths estimated to occur annually in the United States as a result of gram negative bacterial sepsis. Estimates of mortality in patients with profound shock due to endotoxin are 40-50 percent. Recent evidence has implicated cytokines released from endotoxin- stimulated lymphoreticular cells as major factors in the pathogenesis of endotoxin shock. The long range objective of this research project is to define the mechanism(s) by which LPS triggers the activation of lymphoreticular cells with specific focus on endotoxin receptors. For many of these studies, advantage will be taken of the mutant inbred C3H/HeJ mouse strain, whose lymphoreticular cells are refractory to stimulation with many LPS preparations. In this renewal application experiments are described to provide molecular, biochemical and immunologic characterization of a recently identified 80 kDa LPS-specific binding protein expressed on lymphoreticular cells of many species. We will investigate the hypothesis that this 80 kDA candidate LPS receptor serves an important biochemical function in cell activation. Monoclonal antibodies to this protein will be utilized to dissect specific LPS-receptor interactions, to follow the intracellular fate of the receptor and as affinity reagents to purify this molecule. Biochemical characterization and molecular cloning of the structural gene for this protein will be employed to understand further the function of this protein. Experiments to dissect the molecular basis for R-chemotype LPS activation of C3H/HeJ lymphoreticular cells will continue. It is suggested that the successful completion of these studies will contribute to our understanding of molecular interactions of LPS with host cells, and provide information of value in the design of therapeutically effective reagents to treat endotoxin shock.

细菌内毒素脂多糖（LPS）可引发发热， 弥散性血管内重度低血压（休克） 凝血、多系统器官衰竭和死亡。 有 支持内毒素诱导宿主反应的大量证据 是造成25,000多人死亡的主要原因 估计每年发生在美国，由于 革兰氏阴性菌败血症 死亡率估计数 内毒素引起的深度休克患者占40- 50%。 最近的证据表明内毒素释放的细胞因子- 刺激的淋巴网状细胞作为主要因素， 内毒素休克的发病机制。 长期目标： 本研究项目旨在确定LPS 触发淋巴网状细胞的激活， 专注于内毒素受体 对于这些研究中的许多， 将利用突变近交C3 H/HeJ小鼠 淋巴网状细胞对刺激不敏感的菌株 许多LPS制剂。 在此更新应用实验中，描述了提供 A的分子、生化和免疫学特征 最近鉴定的80 kDa LPS特异性结合蛋白表达 许多物种的淋巴网状细胞上。 我们将调查 假设这个80 kDA的候选LPS受体作为 在细胞活化中重要生化功能。 单克隆 针对这种蛋白质的抗体将被用于解剖特定的 LPS-受体相互作用，以跟踪 受体和作为亲和试剂来纯化该分子。 的生化特性和分子克隆 该蛋白质结构基因将用于理解 进一步研究这种蛋白质的功能。 实验来剖析 R-化学型LPS激活C3 H/HeJ的分子基础 淋巴网状细胞将继续存在。 建议由 成功完成这些研究将有助于我们 了解LPS与宿主细胞的分子相互作用， 并提供有价值的信息， 治疗内毒素休克的有效试剂。