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Am J Physiol Cell Physiol (February 6, 2002). doi:10.1152/ajpcell.00385.2001
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Articles in PresS, published online ahead of print February 6, 2002
Am J Physiol Cell Physiol, 10.1152/ajpcell.00385.2001
Submitted on August 9, 2001
Accepted on February 1, 2002

A Role for {delta}-PKC and PI 3-kinase in TNF{alpha} Mediated Anti-Apoptotic Signaling in the Human Neutrophil

Laurie E Kilpatrick1*, Julia Y Lee2, Kathleen M Haines1, Donald E Campbell3, Kathleen E Sullivan3, and Helen M Korchak3

1 Joseph Stokes Research Institute, Children's Hospital of Philadelphia, Philadelphia, PA, USA
2 Pediatrics, University of Pennsylvania, Philadelphia, PA, USA
3 Pediatrics, University of Pennsylvania, Philadelphia, PA, USA; Joseph Stokes Research Institute, Children's Hospital of Philadelphia, Philadelphia, PA, USA

* To whom correspondence should be addressed. E-mail: kilpatrick{at}email.chop.edu.

The proinflammatory cytokine TNF{alpha} has been implicated in the attenuation of neutrophil spontaneous apoptosis during sepsis. Anti-apoptotic signaling is principally mediated through the p60TNF receptor (p60TNFR). In neutrophils, TNF{alpha} is an incomplete secretagogue and requires input from ligated integrin(s) for neutrophil activation. In adherent neutrophils, TNF{alpha} triggered association of both {delta}-PKC and PI 3-kinase with the p60TNFR. In this study, a role for {delta}-PKC and PI 3-kinase in TNF{alpha} mediated anti-apoptotic signaling was examined. TNF{alpha} inhibited spontaneous apoptosis in fibronectin adherent neutrophils and this anti-apoptotic signaling was blocked by the {delta}-PKC inhibitor rottlerin, but not by an inhibitor of Ca-dependent PKC isotypes, Go 6976. Inhibition of PI 3-kinase by LY 294002 also inhibited TNF{alpha} mediated anti-apoptotic signaling. Cycloheximide blocked TNF{alpha} mediated anti-apoptotic signaling suggesting protein synthesis is required. Inhibition of either {delta}-PKC or PI 3-kinase attenuated TNF{alpha}-mediated activation of the anti-apoptotic transcription factor NF{kappa}B. Thus, both {delta}-PKC and PI 3-kinase have essential roles in TNF{alpha} mediated anti-apoptotic signaling in adherent neutrophils.




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