OSTEOCLASTS & GIANT CELLS--IMPLICATIONS OF CELL FUSION

破骨细胞

• 批准号: 3071314
• 负责人: Agnes Vignery
• 金额: $ 5.59万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1987
• 资助国家: 美国
• 起止时间: 1987-01-01 至 1991-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3071314
• 关键词: calcium metabolism; cell fusion; cell membrane; electron microscopy; host organism interaction; laboratory mouse; lymphocyte; macrophage; membrane proteins; monoclonal antibody; osteoclasts; peptidases; physiologic bone resorption; reticuloendothelial system; surface antigens

The long term goals of this proposal are: 1) to elucidate the mechanisms and the regulation of the fusion on mononuclear phagocytes (MNP) leading to the formation of both osteoclasts (OC) and giant cells (GC) and 2) to characterize the function consequences of multinucleation in both normal and pathological bone resorption and in the host defense mechanisms involved in inflammatory reactions and tumors. Two approaches will be used: The first approach focuses upon the cellular and molecular basis for the particular tendency of cells of the MNP lineage to form polykaryons. This approach will more specifically involve kinetic studies of plasma membrane proteins of MNPs induced to form multinucleated cells in vitro, looking for a potential fusogenic protein(s). Combining the specific iodination of plasma membrane proteins, the metabolic labeling of membrane proteins and their subsequent immunoprecipitation with anti-mouse macrophage antibodies we will compare the membrane protein composition of MNPs and GCs, i.e., before and after fusion. The second approach concentrates upon the role of the local microenvironment in the induction and regulation of the formation of multinucleated OC and GC from precursors of the MNP lineage. This approach will more specifically involve studies on the role of soluble factor(s) resulting from lymphocyte- macrophage interactions (Macrophage Fusion Factor). Attempts will be made to purify this putative factor and to analyze which cell(s) and cell interaction(s) lead to its secretion in vitro using monoclonal antibodies directed against specific lymphocyte subsets and macrophages. The effects of local factors such as acidification, proteases and calcium concentration on fusion of MNPs will also be studied in vitro. These two approaches closely interact in that the effects of local factors on the plasma membrane protein composition will also be studied. Parallel studies will be made on in vivo induced OCs and GCs, studying the kinetics of the formation of these cells as well as the molecular composition of their membranes using immunoelectron microscopic techniques and known surface antigen markers. In the long run, these studies may be of critical importance in the understanding of the functional implications of fusion in bone resorption and host defense mechanisms.

这项建议的长期目标是：1)阐明 单核细胞融合的机制及其调控 吞噬细胞(MNP)导致两种破骨细胞的形成 (OC)和巨细胞(GC)和2)以表征功能 正常和病理性多核病变的后果 骨吸收及其在宿主防御机制中的作用 炎症反应和肿瘤。 将使用两种方法：第一种方法侧重于 细胞的特殊倾向的细胞和分子基础 形成多核体的MNP谱系。这种方法将会更多 特别涉及质膜蛋白的动力学研究 体外诱导形成多核细胞的MNPs，寻找 潜在的融合蛋白(S)。结合特定的 质膜蛋白的碘化、代谢性标记 膜蛋白及其随后的免疫沉淀 利用抗鼠巨噬细胞抗体，我们将比较 MNPs和GCs的膜蛋白组成，即前和 在融合之后。 第二种方法集中于地方政府的作用。 微环境在诱导和调节中的作用 多核有机碳和碳氢化合物的形成 MNP血统。这种方法将更具体地涉及到研究 淋巴细胞产生的可溶性因子(S)的作用 巨噬细胞相互作用(巨噬细胞融合因子)。尝试 将致力于提纯这一假定因素，并分析 细胞(S)和细胞相互作用(S)导致其体外分泌 针对特异性淋巴细胞的单抗 亚群和巨噬细胞。地方因素的影响，如 酸化、蛋白水解酶和钙离子浓度对融合率的影响 MNPs也将在体外进行研究。 这两种方法密切互动，因为当地的影响 影响质膜蛋白组成的因素也将是 学习。 将对体内诱导的OCS和GCs进行平行研究， 研究这些细胞形成的动力学以及 用免疫电子方法研究它们膜的分子组成 显微技术和已知的表面抗原标记。 从长远来看，这些研究可能对 理解骨融合的功能含义 再吸收和宿主防御机制。