The survival effect of TNF‐α in human neutrophils is mediated via NF‐κB‐dependent IL‐8 release

Abstract

The capacity of cytokines to modulate neutrophil apoptosis is thought to be a major factor influencing the resolution of granulocytic inflammation. We have previously shown that the late survival effect of TNF-α in human neutrophils involves activation of both NF-κB and phosphoinositide 3-kinase (PI3-kinase) pathways. In this study, we address how these pathways integrate to prevent cell death. In human neutrophils, TNF-α (200 U/ml) induced rapid IκB-α degradation, NF-κB activation and IL-8 release (31.8±5.4 pg/10⁵ cells/2 h), whereas GM-CSF (10 ng/ml) stimulated an equivalent IL-8 release (26.5±4.5 pg/10⁵ cells/2 h) without enhanced IκB-α degradation or NF-κB activation compared to control. Importantly, inhibition of PI3-kinase did not modify TNF-α -induced IκB-α degradation, yet fully inhibited the survival effect of both cytokines. Inhibition of IκB-α phosphorylation, PI3-kinase or ERK1/2 activation blocked IL-8 release by both cytokines. Blocking IL-8 activity by inhibiting its synthesis or by using a neutralizing antibody enhanced the early pro-apoptotic effectof TNF-α and inhibited its late survival effect without affecting GM-CSF-induced survival. These data suggest that cross-talk between NF-κB and PI3-kinase pathways in TNF-α -stimulated neutrophils results from NF-κB/ERK1/2-dependent IL-8 production which acts in an autocrine manner to drive PI3-kinase-dependent survival. In contrast, GM-CSF-mediated survival does not involve NF-κB activation or IL-8 release.