Lu L, Walker D, Graham C D, Waheed A, Shadduck R K, Broxmeyer H E
Department of Medicine, Indiana University School of Medicine, Indianapolis 46223.
Blood. 1988 Jul;72(1):34-41.
The influence of purified recombinant human tumor necrosis factor-alpha (rhuTNF-alpha) was assessed alone and in combination with purified recombinant human interferon gamma (rhuIFN-gamma) for its effects on enhancing release from human monocytes of activities that stimulate colony formation by granulocyte-macrophage (CFU-GM), erythroid (BFU-E), and multipotential (CFU-GEMM) progenitor cells. RhuTNF-alpha or rhuIFN-gamma enhanced release of colony stimulating factors (CSFs), which were determined by a combination of human and mouse colony assays, morphological assessment of colony types and neutralization studies with anti-human macrophage CSF (CSF-1) and anti-human granulocyte (G)-CSF to be CSF-1 and G-CSF. The activity in the uninduced and induced monocyte conditioned media (CM) for CFU-GM-type colonies and clusters was attributed to the presence of both CSF-1 and G-CSF, while the activity in the monocyte CM for BFU-E and CFU-GEMM colonies was attributed to the presence of G-CSF. Monocytes were separated by two-color fluorescence using a dye laser flow cytometry system with cells labeled with anti-leu M3 conjugated with fluorescein isothiocyanate and anti-HLA-DR conjugated with phycoerythrin. While "constitutive" release of CSFs from monocytes was apparent from both the leu M3+, HLA-DR+ and the leu M3+, HLA-DR- (low density or negative DR) fractions, enhanced release of CSFs in response to rhuTNF-alpha or rhuIFN-gamma was confined to the leu M3+, HLA-DR+ population of cells. RhuTNF-alpha and rhuIFN-gamma synergized to enhance release of CSFs such that low concentrations of each molecule, which were inactive when used alone, were active when the two molecules were used together. These studies suggest a role, at least in vitro, for TNF-alpha and IFN-gamma in the release of CSFs from cells of the mononuclear phagocytic lineage.
评估了纯化的重组人肿瘤坏死因子-α（rhuTNF-α）单独及与纯化的重组人干扰素-γ（rhuIFN-γ）联合使用时，对增强人单核细胞释放刺激粒细胞-巨噬细胞（CFU-GM）、红系（BFU-E）和多能（CFU-GEMM）祖细胞集落形成活性的影响。通过人源和鼠源集落测定、集落类型的形态学评估以及用抗人巨噬细胞集落刺激因子（CSF-1）和抗人粒细胞（G）-CSF进行的中和研究相结合，确定rhuTNF-α或rhuIFN-γ增强了集落刺激因子（CSF）的释放，这些集落刺激因子为CSF-1和G-CSF。未诱导和诱导的单核细胞条件培养基（CM）中CFU-GM型集落和集簇的活性归因于CSF-1和G-CSF的存在，而单核细胞CM中BFU-E和CFU-GEMM集落的活性归因于G-CSF的存在。使用染料激光流式细胞术系统通过双色荧光分离单核细胞，细胞用异硫氰酸荧光素偶联的抗-leu M3和藻红蛋白偶联的抗-HLA-DR标记。虽然单核细胞中CSF的“组成性”释放从leu M3 +、HLA-DR +和leu M3 +、HLA-DR -（低密度或阴性DR）组分中都很明显，但rhuTNF-α或rhuIFN-γ诱导的CSF增强释放仅限于leu M3 +、HLA-DR +细胞群体。rhuTNF-α和rhuIFN-γ协同增强CSF的释放，使得单独使用时无活性的低浓度每种分子，当两种分子一起使用时具有活性。这些研究表明，至少在体外，TNF-α和IFN-γ在单核吞噬细胞系细胞释放CSF中发挥作用。
