Józefowicz-Okonkwo Grazyna, Nowak Dariusz
Institute of Physiology and Biochemistry, Medical University of Łódź, Łódź, Poland.
Arch Immunol Ther Exp (Warsz). 2004 Nov-Dec;52(6):447-56.
Previous ex vivo experiments by others suggest that elevated body temperature can prime the respiratory burst of human neutrophils. The mechanism of the priming phenomenon induced by temperature has not been addressed so far. Furthermore, the priming temperature range was not defined.
MATERIAL/METHODS: In the present study we explored ,under in vitro conditions, the influence of febrile-range temperatures on reactive oxygen species (ROS)generation by human peripheral blood neutrophils. ROS production was measured using whole.blood luminol-dependent chemiluminescence. Two elements of signal transduction pathways, calcium and p38 mitogen.activated protein kinase alpha (p38MAPKalpha) ,frequently underlying neutrophil priming were also examined. Calcium levels in the cytosol of resting and fMLP. stimulated isolated neutrophils were measured with the Fura-2AM spectrofluorimetric method. The activity of p38MAPKalpha was assessed indirectly with a specific inhibitor of the kinase, SB 203580.
The study revealed a priming effect at 38 degrees C toward human peripheral blood neutrophil ROS production. Any concomitant effect on calcium response was not observed. Instead, experiments with SB 203580,a specific inhibitor of p38MAPKalpha, pointed to an increased activity of the kinase as a molecular background of temperature-induced priming. However, the priming effect of temperature was confined to 38 degrees C, while higher temperatures proved to exert no effect (39 and 40 degrees C)or even inhibited ROS generation by neutrophils (43 degrees C).
Our study suggests a heterogeneous influence of temperature on human neutrophil functioning, including the priming of the cells by a low-febrile-range temperature. It also suggests a p38MAPKalpha-dependent molecular background of the priming phenomenon.
其他人之前的体外实验表明,体温升高可引发人类中性粒细胞的呼吸爆发。迄今为止,尚未探讨温度诱导引发现象的机制。此外,引发温度范围也未明确。
材料/方法:在本研究中,我们在体外条件下探究了发热范围内的温度对人外周血中性粒细胞产生活性氧(ROS)的影响。使用全血鲁米诺依赖性化学发光法测量ROS的产生。还检测了信号转导途径的两个要素——钙和p38丝裂原活化蛋白激酶α(p38MAPKα),它们是中性粒细胞引发的常见潜在因素。用Fura-2AM荧光分光光度法测量静息和fMLP刺激的分离中性粒细胞胞质溶胶中的钙水平。用激酶的特异性抑制剂SB 203580间接评估p38MAPKα的活性。
该研究揭示了38℃对人外周血中性粒细胞ROS产生的引发作用。未观察到对钙反应的任何伴随影响。相反,使用p38MAPKα的特异性抑制剂SB 203580进行的实验表明,该激酶活性增加是温度诱导引发的分子背景。然而,温度的引发作用仅限于38℃,而更高的温度(39℃和40℃)无作用,甚至会抑制中性粒细胞产生ROS(4