Zsebo K M, Yuschenkoff V N, Schiffer S, Chang D, McCall E, Dinarello C A, Brown M A, Altrock B, Bagby G C
Amgen, Thousand Oaks, CA 91320.
Blood. 1988 Jan;71(1):99-103.
Cultured mononuclear phagocytes produce soluble factors that stimulate endothelial cells to release GM-colony-stimulating activity (GM-CSA). One such factor was recently identified as interleukin 1 (IL 1). Studies were designed to determine which types of granulopoietic factors are released by IL 1-stimulated endothelial cells. Supernatants from endothelial cells cultured for 3 days in medium containing IL 1 alpha and beta were tested in both murine and human CFU-GM colony growth assays. The effect of conditioned media on differentiation of WEHI-3B myelomonocytic leukemic cells was also examined. Control media containing IL 1 alone or unstimulated endothelial cell-conditioned media contained no detectable CSA in any bioassay. Medium conditioned by IL 1-stimulated endothelial cells stimulated the clonal growth of both human and murine CFU-GM and induced macrophage differentiation of WEHI-3B cells. Treatment of these conditioned media with a highly specific neutralizing monoclonal G-CSF antibody completely inhibited their activity in the murine CFU-GM assay, but only partially inhibited GM colony growth by human marrow. Treatment of the active conditioned media with a neutralizing rabbit anti-human GM-CSF antibody partially reduced the activity of the media in the human GM-colony growth assay. G-CSF radioimmunoassay of endothelial cell culture supernatants and Northern blot analysis of endothelial cell cytoplasmic RNA for GM-CSF gene transcripts confirmed that IL 1 induced expression of both G-CSF and GM-CSF genes. Because treatment of media with both antibodies abrogated all activity in the human GM colony growth assay, we conclude that IL 1-stimulated endothelial cells release both G and GM-CSF and that these are the only granulopoietic factors detectable in clonogenic assays released by these cells in vitro.
培养的单核吞噬细胞产生可刺激内皮细胞释放GM集落刺激活性（GM-CSA）的可溶性因子。其中一种因子最近被鉴定为白细胞介素1（IL-1）。本研究旨在确定IL-1刺激的内皮细胞释放哪些类型的粒细胞生成因子。在含有IL-1α和β的培养基中培养3天的内皮细胞上清液，在小鼠和人类CFU-GM集落生长试验中进行检测。还研究了条件培养基对WEHI-3B骨髓单核细胞白血病细胞分化的影响。单独含有IL-1的对照培养基或未刺激的内皮细胞条件培养基在任何生物测定中均未检测到CSA。IL-1刺激的内皮细胞条件培养基刺激了人类和小鼠CFU-GM的克隆生长，并诱导了WEHI-3B细胞的巨噬细胞分化。用高度特异性的中和单克隆G-CSF抗体处理这些条件培养基，完全抑制了它们在小鼠CFU-GM试验中的活性，但仅部分抑制了人类骨髓的GM集落生长。用中和兔抗人GM-CSF抗体处理活性条件培养基，部分降低了培养基在人类GM集落生长试验中的活性。内皮细胞培养上清液的G-CSF放射免疫测定和内皮细胞细胞质RNA的GM-CSF基因转录本的Northern印迹分析证实，IL-1诱导了G-CSF和GM-CSF基因的表达。由于用两种抗体处理培养基消除了人类GM集落生长试验中的所有活性，我们得出结论，IL-1刺激的内皮细胞释放G-CSF和GM-CSF，并且这些是在体外这些细胞释放的克隆形成试验中唯一可检测到的粒细胞生成因子。
