Sieff C A, Niemeyer C M, Mentzer S J, Faller D V
Division of Hematology and Pediatric Oncology, Children's Hospital, Boston, MA 02115.
Blood. 1988 Oct;72(4):1316-23.
Although the genes for four hematopoietic colony-stimulating factors (CSFs) have been cloned, neither the mechanism of the regulation of their production nor their cellular origins have been established with certainty. Monocytes are known to produce colony-stimulating and burst-promoting activities, as well as several monokines such as interleukin-1 (IL-1) and tumor necrosis factor (TNF). These monokines indirectly stimulate other mesenchymal cells to produce certain colony-stimulating factors such as granulocyte-macrophage CSF (GM-CSF). To determine whether monocytes produce other CSFs and if so, to compare the mechanism of regulation of production with that of endothelial cells and fibroblasts, we investigated the synthesis of CSFs by monocytes, human umbilical vein endothelial cells, and fibroblasts. We used total cellular RNA blot analysis to determine interleukin-3 (IL-3), GM-CSF, granulocyte CSF (G-CSF), and monocyte CSF (M-CSF) messenger RNA (mRNA) content and immunoprecipitation or bioassay to confirm the presence of the specific secreted proteins. The results indicate that M-CSF mRNA and protein are produced constitutively by all three cell types and their level of expression does not increase after induction. In contrast, GM-CSF and G-CSF mRNAs are barely detectable in uninduced monocytes and show an increase in expression after lipopolysaccharide treatment. Retrovirus-immortalized endothelial cells, unlike primary endothelial cells or both primary and immortalized fibroblasts, produce IL-1 constitutively; this correlates with their constitutive production of GM-CSF and G-CSF. IL-3 mRNA was not detectable in any of these cells either before or after induction. The results indicate that these mesenchymal cells can produce three CSFs: GM-CSF, G-CSF, and M-CSF; furthermore, the data suggest that the mechanism of regulation of M-CSF production is different from that of GM-CSF and G-CSF, and that the latter two inducible CSFs are regulated by different factors in monocytes compared with the other mesenchymal cells.
尽管四种造血集落刺激因子（CSF）的基因已被克隆，但它们产生的调控机制及其细胞起源均尚未完全明确。已知单核细胞可产生集落刺激活性和爆式促进活性，以及几种单核因子，如白细胞介素-1（IL-1）和肿瘤坏死因子（TNF）。这些单核因子可间接刺激其他间充质细胞产生某些集落刺激因子，如粒细胞-巨噬细胞集落刺激因子（GM-CSF）。为了确定单核细胞是否产生其他CSF，以及如果产生的话，比较其产生的调控机制与内皮细胞和成纤维细胞的调控机制，我们研究了单核细胞、人脐静脉内皮细胞和成纤维细胞对CSF的合成。我们使用总细胞RNA印迹分析来确定白细胞介素-3（IL-3）、GM-CSF、粒细胞集落刺激因子（G-CSF）和单核细胞集落刺激因子（M-CSF）信使RNA（mRNA）的含量，并通过免疫沉淀或生物测定来确认特定分泌蛋白的存在。结果表明，所有三种细胞类型均组成性地产生M-CSF mRNA和蛋白，且其表达水平在诱导后并未增加。相比之下，在未诱导的单核细胞中几乎检测不到GM-CSF和G-CSF mRNA，而在脂多糖处理后其表达增加。与原代内皮细胞或原代及永生化成纤维细胞不同，逆转录病毒永生化的内皮细胞组成性地产生IL-1；这与其组成性产生GM-CSF和G-CSF相关。在诱导前后，这些细胞中均未检测到IL-3 mRNA。结果表明，这些间充质细胞可产生三种CSF：GM-CSF、G-CSF和M-CSF；此外，数据表明M-CSF产生的调控机制与GM-CSF和G-CSF不同，并且与其他间充质细胞相比后两者诱导性CSF在单核细胞中受不同因子调控。
