ALVEOLAR MACROPHAGE - SURFACTANT INTERACTIONS

肺泡巨噬细胞-表面活性剂相互作用

• 批准号: 3087391
• 负责人: ELIZABETH A RICH
• 金额: $ 7.31万
• 依托单位: CASE WESTERN RESERVE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1987
• 资助国家: 美国
• 起止时间: 1987-09-30 至 1992-09-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3087391
• 关键词: affinity chromatography; alveolar macrophages; autoradiography; binding proteins; cell cell interaction; density gradient ultracentrifugation; diagnostic respiratory lavage; electron microscopy; enzyme linked immunosorbent assay; enzyme mechanism; gel electrophoresis; human subject; immunologic techniques; immunoprecipitation; immunoregulation; immunosuppression; laboratory mouse; laboratory rat; lipids; lymphocyte; phosphatidylcholines; protein metabolism; pulmonary surfactants; radioimmunoassay; receptor; thin layer chromatography; tissue /cell culture

Alveolar macrophages (AM) are unique among macrophages in their mitotic potential, use of aerobic metabolism during phagocytosis, and pattern of immunoregulatory function. The factors leading to such specialization are poorly understood. These cells are embedded within surfactant and surfactant affects both effector and immunoregulatory functions of AM. Determination of the mechanisms by which surfactant alters AM function will require scrutiny of the biochemical events occurring during interaction of surfactant and cells. These studies focus on the effect of surfactant on suppressive activities of AM and will test the following hypotheses: surfactant binds to AM via specific surface apoprotein receptors and is distributed to lysosomes where enzymatic reactions lead to its metabolism to amino acids and fatty acids; binding, uptake and/or metabolism of surfactant by human AM contributes to the immunosuppressive activity of AM through the following: a.) enhancement of inhibition by plasma membranes, b.) production of oxidized lipids which are inhibitory, c.) and/or mediators. The hypotheses will be tested according to the following Specific Aims: 1.) to determine the in vitro binding and kinetics of uptake of surfactant by rat AM; 2.) to determine the cellular distribution of surfactant and dipalmitoyl phosphatidylcholine (DPPC) after uptake by rat AM; 3.) to determine the metabolic fate of surfactant after uptake by rat AM and the concomitant release of immunologically active mediators; and 4.) to examine the relationships between surfactant metabolism by human AM and immunoregulation by human AM of lymphocyte response to soluble stimuli. The long- term goal is to apply these findings to disease. Methodology includes cellular immunology (lymphocyte blastogenesis, assay of interleukin-1 activity, enzyme-linked immunoassay) and biochemistry (enzyme assays, assay of 125 Iodine binding and uptake, subcellular fractionation, electrophoresis, lipid analysis).

肺泡巨噬细胞（AM）在巨噬细胞中是独一无二的 它们的有丝分裂潜力，使用有氧代谢 吞噬作用和免疫调节功能的模式。 这 导致这种专业化的因素知之甚少。 这些细胞嵌入在表面活性剂中，表面活性剂影响 AM的效应子和免疫调节功能。 确定表面活性剂改变AM的机制 功能将需要审查发生的生化事件 在表面活性剂和细胞相互作用期间。 这些研究重点 表面活性剂对AM的抑制活动的影响 测试以下假设：表面活性剂通过特定与AM结合 表面载蛋白受体，分布在溶酶体上 酶促反应导致其代谢对氨基酸 和脂肪酸；表面活性剂的结合，摄取和/或代谢 人类AM有助于免疫抑制活性 通过以下内容：a。）增强抑制作用 质膜，b。）氧化脂质的产生 抑制作用，c。）和/或介体。 假设将进行测试 根据以下特定目的：1。）确定 大鼠AM对表面活性剂摄取的体外结合和动力学； 2.） 确定表面活性剂的细胞分布和 大鼠AM摄取后，二氨酰磷脂酰胆碱（DPPC）； 3.）确定摄取后表面活性剂的代谢命运 大鼠AM和免疫学活跃的伴随释放 调解人； 4.）检查 人类AM的表面活性剂代谢和免疫调节 对可溶性刺激的淋巴细胞反应的人类AM。 长期 术语目标是将这些发现应用于疾病。 方法论 包括细胞免疫学（淋巴细胞囊泡发生，测定法 白介素-1活性，酶联免疫测定）和 生物化学（酶测定，测定125种碘结合和 吸收，亚细胞分级，电泳，脂质分析）。