Lindemann A, Riedel D, Oster W, Mertelsmann R, Herrmann F
Department of Hematology, Johannes Gutenberg University, Mainz, FRG.
Eur J Immunol. 1988 Mar;18(3):369-74. doi: 10.1002/eji.1830180308.
Colony-stimulating factors are required for survival proliferation, differentiation and functional activation of granulocytes, macrophages and their precursor cells. In the present report, however, we demonstrate antiproliferative activity of recombinant human (rh) granulocyte-macrophage colony-stimulating factor (GM-CSF) on monoblast cell line U-937 and provide evidence for the involvement of tumor necrosis factor alpha TNF-alpha and interleukin 1 beta (IL 1 beta) in its growth inhibitory action. GM-CSF (but not granulocyte CSF, G-CSF or macrophage CSF, M-CSF) suppressed DNA synthesis and self renewal of U-937 cells. Similarly, medium conditioned by U-937 cells in response to GM-CSF (GM-CSF U-937-CM) was able to reduce clonogenicity and [3H]thymidine uptake by U-937 cells. Since neutralization of GM-CSF present in GM-CSF U-937-CM by monoclonal antibody to GM-CSF did not abrogate the autoinhibitory activity present in GM-CSF U-937-CM, we considered the possibility that other soluble molecules are released by U-937 cells upon GM-CSF stimulation. Neutralization by antibodies to IL 1 beta and TNF-alpha suggested that both monokines could be the antiproliferative principle operating in GM-CSF U-937-CM. Moreover, employing IL 1 beta-specific enzyme-linked immunosorbent assay, TNF-alpha specific radioimmunoassay, Northern analysis using a cloned TNF-alpha-specific cDNA and an oligonucleotide probe for IL 1 beta, we demonstrate GM-CSF-inducible IL 1 beta and TNF-alpha gene expression by U-937 cells at the mRNA and protein level. Although M-CSF expression was induced under similar conditions, M-CSF failed to inhibit growth of U-937 cells.
集落刺激因子是粒细胞、巨噬细胞及其前体细胞存活、增殖、分化和功能激活所必需的。然而，在本报告中，我们证明重组人（rh）粒细胞-巨噬细胞集落刺激因子（GM-CSF）对单核母细胞系U-937具有抗增殖活性，并为肿瘤坏死因子α（TNF-α）和白细胞介素1β（IL-1β）参与其生长抑制作用提供了证据。GM-CSF（而非粒细胞集落刺激因子、G-CSF或巨噬细胞集落刺激因子、M-CSF）抑制U-937细胞的DNA合成和自我更新。同样，U-937细胞在GM-CSF刺激下产生的条件培养基（GM-CSF U-937-CM）能够降低U-937细胞的克隆形成能力和[3H]胸苷摄取。由于用抗GM-CSF单克隆抗体中和GM-CSF U-937-CM中存在的GM-CSF并不能消除GM-CSF U-937-CM中存在的自抑制活性，我们考虑了U-937细胞在GM-CSF刺激下释放其他可溶性分子的可能性。用抗IL-1β和TNF-α抗体进行中和表明，这两种单核因子可能是GM-CSF U-937-CM中发挥抗增殖作用的主要成分。此外，采用IL-1β特异性酶联免疫吸附测定、TNF-α特异性放射免疫测定、使用克隆的TNF-α特异性cDNA和IL-1β寡核苷酸探针进行Northern分析，我们在mRNA和蛋白质水平上证明了U-937细胞在GM-CSF诱导下IL-1β和TNF-α基因的表达。尽管在类似条件下诱导了M-CSF的表达，但M-CSF未能抑制U-937细胞的生长。
