Lopez Juan A, Newburger Peter E, Condino-Neto Antonio
Center for Investigation in Pediatrics and Department of Pharmacology, State University of Campinas Medical School, Campinas SP, Brazil.
J Interferon Cytokine Res. 2003 Dec;23(12):737-44. doi: 10.1089/107999003772084851.
The aim of this study was to investigate the effect of interferon-gamma (IFN-gamma) and tumor necrosis factor-alpha (TNF-alpha) on NADPH oxidase activity and gp91-phox gene expression in HL-60 clone 15 cells as they differentiate along the eosinophilic lineage. The results were compared to the eosoniphilic inducers interleukin-5 (IL-5) and butyric acid. IFN-gamma (100 U/ml) and TNF-alpha (1000 U/ml) or IL-5 (200 pM) caused a significant increase in the expression of the eosinophil peroxidase (EPO) and the major basic protein (MBP) genes. Similar results were observed when the cells were cultured with 0.5 mM butyric acid for 5 days. IFN-gamma (100 U/ml) and TNF-alpha (1000 U/ml) also caused a significant increase in superoxide release by HL-60 clone 15 cells after 2 days compared with control or with butyric acid-induced cells. After 5 days, these cytokines and butyric acid induced an even stronger release of superoxide. HL-60 clone 15 cells cultured with IFN-gamma and TNF-alpha for 2 days showed a significant increase in gp91-phox gene expression. We conclude that IFN-gamma and TNF-alpha are sufficient to induce the differentiation of HL-60 clone 15 cells to the eosinophilic lineage and to upregulate gp91-phox gene expression and activity of the NADPH oxidase system.
本研究旨在探讨γ干扰素（IFN-γ）和肿瘤坏死因子-α（TNF-α）对HL-60克隆15细胞沿嗜酸性粒细胞谱系分化过程中NADPH氧化酶活性及gp91-phox基因表达的影响。将结果与嗜酸性粒细胞诱导剂白细胞介素-5（IL-5）和丁酸进行比较。IFN-γ（100 U/ml）、TNF-α（1000 U/ml）或IL-5（200 pM）可显著增加嗜酸性粒细胞过氧化物酶（EPO）和主要碱性蛋白（MBP）基因的表达。当细胞用0.5 mM丁酸培养5天时，观察到类似结果。与对照组或丁酸诱导的细胞相比，IFN-γ（100 U/ml）和TNF-α（1000 U/ml）在2天后也可显著增加HL-60克隆15细胞的超氧化物释放。5天后，这些细胞因子和丁酸诱导的超氧化物释放更强。用IFN-γ和TNF-α培养2天的HL-60克隆15细胞显示gp91-phox基因表达显著增加。我们得出结论，IFN-γ和TNF-α足以诱导HL-60克隆15细胞向嗜酸性粒细胞谱系分化，并上调gp91-phox基因表达及NADPH氧化酶系统的活性。
