MONONUCLEAR PHAGOCYTE-DERIVED GROWTH FACTOR

这项研究的目标是发展我们对 单核巨噬细胞调控增殖的基本机制(S) 各种正常细胞类型。到目前为止，我们的发现表明 巨噬细胞和巨噬细胞样细胞系阐述特异的可溶性 可导致静止的成纤维细胞启动横跨血管的因子(S) 细胞周期。在操作上，这个独角兽(S)是一个“能力”因素， 其作用类似于其他已知的活性因子(血小板源 生长因子和成纤维细胞生长因子)，但在生化上与 两者都有。我们集中力量净化空气中的 人巨噬细胞衍生能力因子(MDCF) 培养的P388D1，一种小鼠巨噬细胞样细胞系，阐述了慷慨的 除另一种生长因子白介素1(IL-1)外，还需补充一定数量的MDCF。 我们通过使P388D1细胞生长来极大地简化了纯化 在规定的介质中连续无血清。的串口应用 超滤、凝胶过滤和离子交换层析 在生化/生物物理方面明确分离的MDCF和IL-1 性质和生物活性。银染十二烷基硫酸钠-PAGE分析 平板凝胶显示了与MDCF相对应的两条主要蛋白质条带 活性和与IL-1活性相对应的一个。的配套研究 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。使用这个大大提纯的MDCF，我们 现在开始用兔子和单抗研制出特异的抗血清 用大鼠-小鼠杂交瘤抗体鉴定和定量mdcf 体内和体外实验。我们的目标是将这只独角兽与 其他已知的生长因子，作为使用这些因子作为探针的第一步 剖析与增殖相关的细胞事件。结果是 所获得的结果应为明确研究两者之间的关系奠定基础 体内巨噬细胞衍生能力因子的产生对损伤的影响 以单核巨噬细胞堆积，广泛纤维增生为特征， 和胶原蛋白沉积。(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)