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and PI 3-kinase
in TNF-
-mediated antiapoptotic signaling in
the human neutrophil
Department of Pediatrics, University of Pennsylvania School of Medicine and the Joseph Stokes Jr. Research Institute, Children's Hospital of Philadelphia, Philadelphia, Pennsylvania 19104
The
proinflammatory cytokine tumor necrosis factor (TNF)-
has been
implicated in the attenuation of neutrophil spontaneous apoptosis during sepsis. Antiapoptotic signaling is
principally mediated through the p60TNF receptor (p60TNFR). In
neutrophils, TNF- is an incomplete secretagogue and requires input
from a ligated integrin(s) for neutrophil activation. In adherent
neutrophils, TNF- triggers association of both protein kinase C
(PKC)-
and phosphatidylinositol (PI) 3-kinase with the p60TNFR. In
this study, a role for PKC- and PI 3-kinase in TNF--mediated
antiapoptotic signaling was examined. TNF- inhibited spontaneous
apoptosis in fibronectin-adherent neutrophils, and this
antiapoptotic signaling was blocked by the PKC- inhibitor
rottlerin, but not by an inhibitor of Ca2+-dependent PKC
isotypes, Go-6976. Inhibition of PI 3-kinase by LY-294002 also
inhibited TNF--mediated antiapoptotic signaling. Cycloheximide
blocked TNF--mediated antiapoptotic signaling, suggesting
protein synthesis is required. Inhibition of either PKC- or PI
3-kinase attenuated TNF--mediated activation of the antiapoptotic transcription factor NF
B. Thus both PKC- and PI 3-kinase have essential roles in TNF--mediated antiapoptotic signaling in adherent neutrophils.
sepsis; inflammation; signal transduction; nuclear factor B; protein kinase C-; tumor necrosis factor-
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