NF-kappa B activation is a critical regulator of human granulocyte apoptosis in vitro

During beneficial inflammation, potentially tissue-damaging granulocytes un dergo apoptosis before being cleared by phagocytes in a non-phlogistic mann er. Here we show that the rate of constitutive apoptosis in human neutrophi ls and eosinophils is greatly accelerated in both a rapid and concentration -dependent manner by the fungal metabolite gliotoxin, but not by its inacti ve analog methylthiogliotoxin, This induction of apoptosis was abolished by the caspase inhibitor zVAD-fmk, correlated with the inhibition of nuclear factor-kappa B (NF-kappa B), and was mimicked by a cell permeable inhibitor y peptide of NF-kappa B, SN-50; other NF-kappa B inhibitors, curcumin and p yrrolidine dithiocarbamate; and the proteasome inhibitor, MG-132, Gliotoxin also augmented dramatically the early (2-6 h) pro-apoptotic effects of tum or necrosis factor-alpha (TNF-alpha) in neutrophils and unmasked the abilit y of TNF-alpha to induce eosinophil apoptosis, In neutrophils, TNF-alpha ca used a gliotoxin-inhibitable activation of an inducible form of NF-kappa B, a response that may underlie the ability of TNF-alpha to delay apoptosis a t later times (12-24 h) and limit its early killing effect. Furthermore, cy cloheximide displayed a similar capacity to enhance TNF-alpha induced neutr ophil apoptosis even at time points when cycloheximide alone had no pro-apo ptotic effect, suggesting that NF-kappa B may regulate the production of pr otein(s) which protect neutrophils from the cytotoxic effects of TNF-alpha, These data shed light on the biochemical and molecular mechanisms regulati ng human granulocyte apoptosis and, in particular, indicate that the transc ription factor NF-kappa B plays a crucial role in regulating the physiologi cal cell death pathway in granulocytes.