MACROPHAGE GROWTH: ROLE OF COLONY STIMULATING FACTOR

正常小鼠腹膜渗出液的生长和增殖 巨噬细胞(PEM)在体外严格依赖于一种巨噬细胞特异性 生长因子称为集落刺激因子(CSF-1)。相反， 转化的巨噬细胞瘤细胞系不需要外源性CSF-1 文化的成长。最近的研究表明，酪氨酸蛋白的磷酸化 某些膜结合蛋白可能参与细胞的控制。 由特定的生长因子诱导的增殖。此外，还有几个 癌基因产物已被证明在结构上与任何一种 特定的生长因子或生长因子受体。 我们建议同时使用正常巨噬细胞和已建立的巨噬细胞。 以具有不同生长表型的品系为模型考察其作用 蛋白磷酸化在巨噬细胞增殖调控中的作用。 首先，我们将发展双向凝胶电泳法和 放射自显影技术识别CSF-1诱导和/或自发的 正常巨噬细胞的磷酸化成分和已建立的 巨噬细胞-单核细胞肿瘤细胞系。薄层平板电泳法 技术将被用来表征化学性质的 磷酸化氨基酸。第二，一种抗人血吸虫单抗 将开发磷酸酪氨酸部分作为探针，并与“Western”一起开发 印迹“技术，以促进鉴定和表征 酪氨酸磷酸化的蛋白质。 我们将研究是否产生自分泌的CSF-1或CSF样 活性和酪氨酸磷酸化与致癌有关 巨噬细胞在发育早期的转化。我们 将通过肿瘤病毒转化建立永生化的小鼠骨 以研究这些细胞系是否能产生和 分泌自我刺激活性。我们还将审查和确定 这些细胞系中的磷酸化成分。我们的假设 工作是自分泌生长因子，酪氨酸磷酸化和 配体-受体相互作用在细胞内起着重要作用 骨髓源性造血细胞的转化和致癌作用。

The growth and proliferation of normal murine peritoneal exudate macrophages (PEM) in vitro is strictly dependent on a macrophage-specific growth factor known as colony-stimulating factor (CSF-1). By contrast, transformed macrophage tumor cell lines do not require exogenous CSF-1 for growth in culture. Recent studies show that tyrosine phosphorylation of certain membrane-bound proteins may be involved in the control of cell proliferation induced by specific growth factors. In addition, several oncogene products have been shown to be structurally related to either specific growth factors or growth factor receptors. We propose to use both normal macrophages and established macrophage cell lines with different growth phenotypes as models to investigate the role of protein phosphorylation in the regulation of macrophage proliferation. First, we will develop two dimensional gel electrophoresis and autoradiography techniques to identify the CSF-1 induced and/or spontaneous phosphorylated components from both mormal macrophages and established macrophage-monocyte tumor cell lines. Thin layer plate electrophoresis techniques will be used to characterize the chemical nature of the phosphorylated amino acids. Second, a monoclonal antibody against a phosphotyrosine moiety will be developed as a probe, along with "Western blot" techniques, to facilitate the identification and characterization of tyrosine-phosphorylated-proteins. We will study whether the production of autocrine CSF-1 or CSF-like activity and tyrosine phosphorylation are linked to oncogenic transformation of macrophages during the early phase of development. We will establish, by tumor virus transformation, immortalized mouse bone marrow-derived macrophages to study whether these cell lines produce and secrete autostimulatory activity. We will also examine and identify phosphorylated components from these cell lines. The hypothesis of our work is that autocrine growth factor, tyrosine phosphorylation and ligand-receptor interaction play important roles in the cellular transformation and oncogenesis of bone marrow-derived hematopoietic cells.