EFFECT OF O3, NO2 ON LUNG MACROPHAGE PHAGOCYTOSIS

O3、NO2对肺巨噬细胞吞噬功能的影响

• 批准号: 3252291
• 负责人: RALPH J PAROD
• 金额: $ 7.68万
• 依托单位: STATE UNIVERSITY OF NEW YORK AT BUFFALO
• 依托单位国家: 美国
• 财政年份: 1986
• 资助国家: 美国
• 起止时间: 1986-07-01 至 1992-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3252291
• 关键词: air pollution; alveolar macrophages; antibody specificity; calcium; cell age; environmental toxicology; flow cytometry; fluorescence microscopy; gas analyzer; gel electrophoresis; hamsters; immunoregulation; inhalation drug administration; leukotrienes; monoclonal antibody; opsonin; ozone; phagocytosis; pollution related respiratory disorder; receptor; respiratory toxin; scintillation counter; scintillation spectrometry; surface antigens

Pulmonary macrophages (PM) aid lung defense by removing a variety of deposited pathogens through the process of phagocytosis. This process begins with the specific interaction of particle surface ligands with ligand receptors in the macrophage cell membrane. Unlike immunologically opsonized pathogens, PM receptors for pathogens which do not generate an immune response remain largely undefined. We have developed new techniques to investigate phagocytosis. These include the ability to distinguish and quantitate bound versus ingested particles as well as the ability to evaluate the heterogeneity of these and other characteristics within the PM population. Using a series of in vitro experiments, we propose to characterize the mechanisms by which particles with different surface charges and particles opsonized with immunoglobulin are phagocytized. Particles will be prepared by coating fluorescent latex beads with appropriate proteins. The quantitation of attached versus ingested particles will be made through the combined use of flow cytometry and fluorescence microscopy. Particles will also be used to isolate the cell surface receptors to which they bind. Antibodies raised against these receptors will allow us to determine if changes in particle binding reflect changes in receptor number or particle- receptor affinity. Another tool we have developed is a monoclonal antibody against an age-related cell surface antigen specific to hamster PM. This antigen may be the receptor for negatively-charged particles. Interestingly, our antibody and calcium ionophore inhibit ingestion, presumably by raising intracellular calcium levels. This contention must be reconciled with the fact that calcium ionophore also stimulates leukotriene synthesis. Particles, our monoclonal antibody, calcium ionophore and metals will be used to test hypothesis concerning the role of calcium and leukotrienes in the mechanism which couples particle-receptor binding to particle internalization. The integration of this information will enhance our understanding of phagocytosis and its cellular rregulation in vivo.

肺巨噬细胞（PM）通过清除肺内的 各种各样的沉积病原体通过的过程中， 吞噬作用 这一过程开始于特定的相互作用， 颗粒表面配体与巨噬细胞中的配体受体 细胞膜 与免疫调理的病原体不同，PM 不产生免疫反应的病原体受体 在很大程度上是不确定的。 我们开发了新技术， 研究吞噬作用。 其中包括区分能力 并定量结合与摄入的颗粒以及 评估这些和其他异质性的能力 PM人群的特征。 使用一系列在 体外实验中，我们建议通过以下方式来表征机制： 具有不同表面电荷的粒子 用免疫球蛋白调理的细胞被吞噬。 颗粒将 通过用适当的荧光剂涂覆荧光乳胶珠来制备 proteins. 附着颗粒与摄入颗粒的定量 将通过联合使用流式细胞术和 荧光显微镜 粒子也将被用来分离 它们所结合的细胞表面受体。 产生的抗体 将使我们能够确定 颗粒结合反映受体数量或颗粒的变化， 受体亲和力 我们开发的另一个工具是 抗年龄相关细胞表面抗原的单克隆抗体 专为仓鼠PM。 这种抗原可能是 带负电荷的粒子 有趣的是，我们的抗体和 钙离子载体抑制摄入，可能是通过提高 细胞内钙水平。 这一论点必须加以调和 事实上钙离子载体也刺激白三烯 合成. 微粒，我们的单克隆抗体，钙离子载体 和金属将被用来测试假设的作用， 钙和白三烯的机制， 粒子-受体结合到粒子内化。 的 这些信息的整合将增强我们对 吞噬作用及其在体内的细胞失调。