CELL SURFACE AND OSTEOCLAST DEVELOPMENT

正常的骨生长和重建部分取决于以下因素： 影响破骨细胞的数量和活性。 骨骼 破骨细胞质膜的改变可能导致病理学改变 或控制破骨细胞分化和再吸收的信号 活动 尽管最近有大量的见解来理解 造血谱系以及局部和系统的相互作用 破骨细胞发育和功能的调节剂，许多问题仍然存在。 此外，成熟的生物化学和分子概况 破骨细胞是不完全的。 本申请所依据的假设 提出破骨细胞的发育需要特定的细胞表面 改变和离散的膜相关分子参与 破骨细胞调节和功能。 具体目标是 建议是：1）进一步表征与所述抗原反应的抗原， 121 F单克隆抗体（MAb）并阐明其与 超氧化物歧化酶，2）继续鉴定和生化描述 不同的破骨细胞质膜分子被一组 破骨细胞特异性单克隆抗体，3）研究细胞表面变化相关 与破骨细胞发育和功能状态有关， 已知的破骨细胞发育和活性的调节剂（如 雌二醇、降钙素、白细胞介素1和6以及TNF）以及诱导的 原癌基因（c-fos、c-jun和c-src）的表达，以及4）鉴定 破骨细胞前体和破骨细胞细胞分化的定义阶段 使用这些细胞表面标记和分子探针。 高度纯化 将使用鸡破骨细胞和人白血病细胞系（FLG 29.1 作为鸡和人类抗原的来源。 在其他实验中，骨髓 单核细胞融合形成多核破骨细胞样巨人 培养中的细胞将作为一个发育系统， 研究已知骨调节剂对抗原表达的影响 和吸收能力，并确定破骨细胞前体。 生物化学、生理学、免疫组织化学和分子克隆 技术将被用来确定的性质和功能作用 这些抗原。 Western和北方印迹分析将用于 研究它们与原癌基因表达的关系。 这些结果 不仅应该扩大我们对细胞的基本理解， 破骨细胞的分子生物学，而且还可能有助于设计 适当的预防和治疗策略， 关节炎、骨质疏松症、牙周病等骨质减少 骨的局部或代谢紊乱。

Normal bone growth and remodeling are, in part, dependent upon factors that influence the number and activity of osteoclasts. Skeletal pathologies may result from alterations in the osteoclast plasma membrane or in signals that govern osteoclast cytodifferentiation and resorptive activity. Despite substantial recent insights into understanding the hematopoietic lineage and the interactions of local and systemic modulators of osteoclast development and function, many questions remain. In addition, the biochemical and molecular profile of the mature osteoclast is incomplete. The hypothesis upon which this application is based proposes that osteoclast development entails specific cell surface alterations and that discrete membrane-associated molecules are involved in osteoclast regulation and function. The specific aims of this proposal are to: 1) further characterize the antigen reactive with the 121F monoclonal antibody (MAb) and to clarify its relationship with superoxide dismutase, 2) continue to identify and biochemically describe distinct osteoclast plasma membrane molecules recognized by a panel of osteoclast-specific MAbs, 3) investigate cell surface changes associated with osteoclast development and functional state, relative to the effects of known modulators of osteoclast development and activity (such as estradiol, calcitonin, interleukins 1 and 6, and TNF) and to the induced expression of protooncogenes (c-fos, c-jun, and c-src), and 4) identify osteoclast precursors and define stages of osteoclast cytodifferentiation using-these cell surface markers and molecular probes. Highly purified chick osteoclasts and a human leukemic cell line (FLG 29.1) will be used as sources for chicken and human antigens. In other experiments, marrow mononuclear cells which fuse to form multinucleated osteoclast-like giant cells in culture will serve as a developmental system in which to investigate the influence of known bone modulators on antigen expression and resorptive capability, and to define osteoclast precursors. Biochemical,physiological, immunohistochemical, and molecular cloning techniques will be employed to ascertain the nature and functional role of these antigens. Western and northern blot analyses will be used to examine their relationship to protooncogene expression. These results should not only expand our basic understanding of the cellular and molecular biology of the osteoclast, but also potentially aid in devising appropriate preventative and therapeutic strategies for the skeletal osteopenia of arthritis, osteoporosis, periodontal disease, and other local or metabolic disorders of bone.