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OSTEOCLASTS & GIANT CELLS: IMPLICATIONS OF CELL FUSION

破骨细胞

基本信息

批准号：
3071315
负责人：
Agnes Vignery
金额：
$ 7.08万
依托单位：
YALE UNIVERSITY
依托单位国家：
美国
项目类别：
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/3071315
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

项目摘要

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.

这项建议的长远目标是：1）阐明单核细胞融合的机制及调控吞噬细胞（MNP）导致破骨细胞的形成 (OC)和巨细胞（GC）和2）表征功能正常和病理性多核化的后果骨吸收和参与宿主防御机制，炎症反应和肿瘤。将使用两种方法：第一种方法侧重于细胞和分子基础的细胞的特定趋势，形成多核体。这种方法将更多具体涉及质膜蛋白质动力学研究的MNPs诱导形成多核细胞在体外，寻找潜在的融合蛋白。结合具体质膜蛋白质的碘化，膜蛋白及其随后的免疫沉淀用抗小鼠巨噬细胞抗体，我们将比较 MNP和GC的膜蛋白组成，即，之前和融合后。第二种方法集中于地方的作用微环境的诱导和调节从多核OC和GC的前体形成 MNP血统。这种方法将更具体地涉及研究对淋巴细胞产生的可溶性因子的作用- 巨噬细胞融合因子（Macrophage Fusion Factor）尝试本文将对这一假定因素进行提纯，并分析细胞和细胞相互作用导致其在体外分泌，特异性淋巴细胞单克隆抗体亚群和巨噬细胞当地因素的影响，如酸化、蛋白酶和钙浓度对融合的影响还将在体外研究MNP。这两种方法密切相互作用，因为当地的影响对质膜蛋白质组成的因素也将是研究了将对体内诱导的OC和GC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显微镜观察细胞膜的分子组成显微镜技术和已知的表面抗原标记。从长远来看，这些研究可能对了解骨融合的功能意义再吸收和宿主防御机制。