MONOCYTE-SPECIFIC CHEMOATTRACTANT FROM HUMAN PMN

来自人类 PMN 的单核细胞特异性趋化剂

• 批准号: 3142325
• 负责人: John K Spitznagel
• 金额: $ 14.8万
• 依托单位: EMORY UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1990
• 资助国家: 美国
• 起止时间: 1990-07-01 至 1994-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3142325
• 关键词: MHC class II antigen; bioassay; cell cell interaction; cell migration; chemical binding; chemoattractants; chemotaxis; genetic mapping; human subject; inflammation; interleukin 1; laboratory mouse; laboratory rabbit; lysozyme; macrophage; molecular cloning; monocyte; neutrophil; receptor; superoxides; tumor necrosis factor alpha

The emigration of neutrophils (Pmns) and monocytes is one of the fundamental events in inflammation. These cells are intimately involved in the elimination of harmful microbes, toxins, and denatured proteins and in the immune reaction to foreign antigens. Pmns are the predominant cell in the acute inflammatory stage, whereas monocytes are found in the chronic stage. The stimulus that mediates the second wave of inflammation resulting in the migration of monocytes into an area has not yet been elucidated. The hypothesis proposed in this study is that this influx of monocytes is in part due to the liberation of a chemoattractant from the neutrophil itself, either following the death of the neutrophil or subsequent to degranulation. The validity of this hypothesis will be determined by the following specific aims. Aim 1, to demonstrate that a naturally occurring cationic granule protein of human Pmns of Mr 37000D (CAP37) has potent chemotactic activity specific for human monocytes, as measured by the modified Boyden chamber technique, and to ask what other responses CAP 37 induces in human monocytes. Aim 2, to determine the amino acid sequence of CAP 37 and the cloning and sequencing of its structural gene so as to define the domains responsible for chemotactic activity. Aim 3, to ask if monocytes have specific receptors that mediate chemotaxis. With radiolabeled proteins we will attempt to detect and measure the saturable and specific binding sites for CAP37 on human monocytes. Aim 4, to evaluate the role of CAP37 as a monocyte- specific chemoattractant in vivo and to ask what other responses CAP37 may exert on monocytes/macrophages in vivo. The release of lysosomal enzymes, IL-1, tumor necrosis factor, superoxide production, and expression of La (class II MHC protein) will be determined with standard assays. It is believed that through these investigations CAP37 will be demonstrated to cause migration of monocytes and modulate other functions in vivo, and that its effects on the activity of monocytes and macrophages at the sites of inflammation will be determined.

中性粒细胞(PMN)和单核细胞的迁移是 炎症中的基本事件。这些细胞密切相关 在消除有害微生物、毒素和变性蛋白质以及 在对外来抗原的免疫反应中。PMN是最主要的细胞 在急性炎症阶段，而单核细胞在 慢性期。调节第二波炎症的刺激因素 导致单核细胞迁移到尚未进入的区域 已被阐明。这项研究中提出的假设是，这 单核细胞的流入部分是由于一种化学诱导剂的释放。 从中性粒细胞本身，无论是在中性粒细胞死亡之后 或在脱颗粒之后。这一假设的正确性将 由以下具体目标决定。目标1，证明 天然存在的人PMN的MR 37000D阳离子颗粒蛋白 (CAP37)对人类单核细胞具有强烈的趋化活性， 通过改进的博伊登小室技术进行测量，并询问 CAP 37在人单核细胞中诱导的其他反应。目标2，确定 CAP 37的氨基酸序列及ITS的克隆和序列分析 结构基因，以确定负责趋化的结构域 活动。目标3，询问单核细胞是否有特异的受体 调节趋化作用。对于放射性标记的蛋白质，我们将尝试检测 并测定了CAP37在人体上的饱和结合部位和特异结合部位 单核细胞。目的4、评价CAP37作为单核细胞的作用。 并询问CAP37可能有什么其他反应 作用于体内单核/巨噬细胞。溶酶体的释放 酶、IL-1、肿瘤坏死因子、超氧化物产生和 La(II类MHC蛋白)的表达将用标准方法测定 化验。相信通过这些调查，CAP37将是 证明能引起单核细胞迁移并调节其他功能 以及对单核细胞和巨噬细胞活性的影响 炎症部位的位置将被确定。