McDonald Patrick P, Ye Richard D
Department of Pharmacology, Univeristy of Illinois,Chicago, IL, USA.
Methods Mol Biol. 2007;412:473-86. doi: 10.1007/978-1-59745-467-4_29.
The crucial contribution of neutrophils to innate immunity extends well beyond their traditional role as professional phagocytes. Indeed, it is now well established that neutrophils generate a plethora of inflammatory cytokines and chemokines that are profoundly involved in the onset and evolution of the inflammatory reaction. Several recent studies have also shown that neutrophils can represent an important source of inflammatory cytokines in a number of pathophysiological settings. The inflammatory cytokines produced by neutrophils are generally encoded by immediate-early response genes, which in turn depend on the activation of transcription factors such as those belonging to the nuclear factor (NF)-kappaB and signal transducers and activators of transcription (STAT) families. We have shown in the past that such factors are expressed and inducible in neutrophils stimulated by physiological agonists. However, the detection of intact (i.e., undegraded) transcription factors in neutrophils requires special precautions and an alternative protocol, as a result of the huge amounts of endogenous proteases present in these cells. This protocol is the focus of this chapter.
中性粒细胞对固有免疫的关键贡献远远超出了它们作为专职吞噬细胞的传统作用。事实上，现在已经明确，中性粒细胞会产生大量炎性细胞因子和趋化因子，这些因子在炎症反应的发生和发展中起着至关重要的作用。最近的几项研究还表明，在许多病理生理情况下，中性粒细胞可能是炎性细胞因子的重要来源。中性粒细胞产生的炎性细胞因子通常由即刻早期反应基因编码，而这些基因又依赖于转录因子的激活，如属于核因子（NF）-κB和信号转导及转录激活因子（STAT）家族的转录因子。我们过去已经表明，这些因子在生理激动剂刺激的中性粒细胞中表达且可诱导。然而，由于这些细胞中存在大量内源性蛋白酶，要在中性粒细胞中检测完整的（即未降解的）转录因子需要特别的预防措施和替代方案。本章将重点介绍这个方案。
