MONONUCLEAR PHAGOCYTE-DERIVED GROWTH FACTOR

这项研究的目标是发展我们对 单核吞噬细胞调节增殖的基本机制 不同类型的正常细胞。 我们迄今为止的发现表明， 巨噬细胞和巨噬细胞样细胞系阐述了特异性可溶性 可引起静止的成纤维细胞开始穿过细胞膜的因子 细胞周期 在操作上，这种单核因子是一种“能力”因素， 其作用类似于其它已知的感受态因子（血小板衍生的 生长因子和成纤维细胞生长因子），但在生物化学上与 两者 我们集中力量净化 巨噬细胞源性能力因子（MDCF），来自 培养的P388 D1是一种小鼠巨噬细胞样细胞系， 量的MDCF以及另一种生长因子白细胞介素-1（IL-1）。 我们通过使P388 D1细胞适应生长， 在限定培养基中连续无血清培养。 系列应用 超滤、凝胶过滤和离子交换色谱法， MDCF和IL-1在生物化学/生物物理学方面均明确分离 性质和生物活性。 银染SDS-PAGE分析 平板凝胶显示两条主要蛋白带，对应于MDCF 活性和对应于IL-1活性的一个。 相关研究 亲和纯化的IL-1证实了我们的发现，MDCF和IL-1是不同的 生化和生物实体。 使用这种高度纯化的MDCF，我们 现在开始使用兔和单克隆抗体开发特异性抗血清， 使用大鼠-小鼠杂交瘤抗体鉴定和定量MDCF， 体内和体外实验。 我们的目的是将这种单核细胞与 作为使用这些作为探针的初始步骤， 剖析与增殖相关的细胞事件。 结果 获得的结果应该为确定性的研究提供基础。 巨噬细胞源性感受态因子在体内对病变的影响 其特征在于单核吞噬细胞积聚，广泛的纤维增生， 和胶原沉积。 （J）

The goal of this research is to develop our fundamental knowledge of the basic mechanism(s) by which mononuclear phagocytes regulate proliferation of various normal cell types. Our findings to date suggest that macrophages and macrophage-like cell lines elaborate specific soluble factor(s) that can cause quiescent fibroblasts to initiate traverse of the cell cycle. Operationally, this monokine(s) is a "competence" factor, similar in its action to other known competence factors (platelet-derived growth factor and fibroblast growth factor) but biochemically distinct from both. We have concentrated our efforts on purification of the macrophage-derived competence factor (MDCF) from spent supernates of cultured P388D1, a mouse macrophage-like cell line that elaborates generous amounts of MDCF in addition to another growth factor, interleukin-1 (IL-1). We have greatly simplified purification by adapting P388D1 cells to grow continuously serum-free in a defined medium. Serial application of ultrafiltration, gel filtration, and ion exchange chromatography has clearly separated MDCF and IL-1, both in terms of biochemical/biophysical properties and biological activities. Analysis by silver-stained SDS-PAGE slab gels reveals two major bands of protein that correspond to MDCF activity and one that corresponds to IL-1 activity. Companion studies of affinity-purified IL-1 confirm our findings that MDCF and IL-1 are distinct biochemical and biological entities. Using this greatly purified MDCF, we are now beginning to develop specific antisera using rabbits and monoclonal antibodies using rat-mouse hybridomas to identify and quantify MDCF for in vivo and in vitro experiments. Our aim is to compare this monokine with other known growth factors as an initial step in using these as probes to dissect the cellular events associated with proliferation. The results obtained should provide a foundation for definitive studies on the relation of macrophage-derived competence factor production in vivo to lesions characterized by mononuclear phagocyte accumulation, extensive fibroplasia, and collagen deposition. (J)