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首页> 外文期刊>Cytokine >GM-CSF and TNFalpha modulate protein expression of human neutrophils visualized by fluorescence two-dimensional difference gel electrophoresis.
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GM-CSF and TNFalpha modulate protein expression of human neutrophils visualized by fluorescence two-dimensional difference gel electrophoresis.

机译：GM-CSF和TNFalpha调节通过荧光二维差异凝胶电泳可视化的人类嗜中性粒细胞的蛋白表达。

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Increased serum levels of TNFalpha and GM-CSF are found in various chronic inflammatory diseases and these cytokines affect the function of circulating and tissue neutrophils. TNFalpha- and GM-CSF-induced protein expression profiles could, therefore, serve as biomarker for the action of these cytokines in vivo. We stimulated human peripheral neutrophils with TNFalpha and GM-CSF in vitro and analyzed changes in their proteome by fluorescence two-dimensional difference gel electrophoresis (2D-DIGE). We report the differential expression of 3 and 18 protein spots following TNFalpha and GM-CSF stimulation, respectively. Differences in protein expression induced by TNFalpha were limited and did not show discriminatory power in a principal component analysis, whereas the profile induced by GM-CSF did. TNFalpha- and GM-CSF-induced both de novo IL-1beta and sIL-1Ra protein expression as detected by Western blot analysis, which confirmed proper neutrophil activation by these cytokines in vitro. Mass spectrometry analysis of cytokine-regulated protein spots resulted in the identification of 8 proteins. Among the identified proteins, enolase 1 and annexin A1 might function as markers for peripheral neutrophil activation. In conclusion, a proteomic analysis of neutrophils by 2D-DIGE provides proof-of-principle that cytokine-induced protein profiles can serve as biomarkers for the action of individual cytokines in vivo.
机译：在各种慢性炎性疾病中发现血清TNFalpha和GM-CSF的水平升高，这些细胞因子影响循环和组织中性粒细胞的功能。因此，TNFalpha和GM-CSF诱导的蛋白表达谱可作为这些细胞因子在体内作用的生物标记。我们在体外用TNFalpha和GM-CSF刺激人外周血中性粒细胞，并通过荧光二维差异凝胶电泳（2D-DIGE）分析了它们的蛋白质组变化。我们报告分别在TNFalpha和GM-CSF刺激后3和18蛋白点的差异表达。由TNFα诱导的蛋白质表达差异有限，在主成分分析中没有显示出鉴别力，而由GM-CSF诱导的表达谱却有区别。通过蛋白质印迹分析检测到，TNFalpha和GM-CSF诱导了从头的IL-1beta和sIL-1Ra蛋白表达，这证实了这些细胞因子在体外能正确激活嗜中性粒细胞。质谱分析细胞因子调节的蛋白质斑点可鉴定出8种蛋白质。在鉴定出的蛋白质中，烯醇化酶1和膜联蛋白A1可能充当外周嗜中性粒细胞活化的标志物。总之，通过2D-DIGE对嗜中性粒细胞进行蛋白质组学分析的原理证明，细胞因子诱导的蛋白质谱可作为体内单个细胞因子作用的生物标记。

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