Protein kinase C activation modulates tumour necrosis factor-alpha priming of human neutrophils for zymosan-induced leukotriene B4 release.
Neutrophil (PMN) activation by the yeast component zymosan involves the complement receptor type 3 (CD11b/CD18). Recombinant human tumour necrosis factor-alpha (rhTNF-alpha) augmented the zymosan-stimulated leukotriene B4 (LTB4) release from PMN, reaching a fourfold increase at 10(-9) M. Co-incubation of PMN with 10(-9) M rhTNF-alpha and staurosporine resulted in a further dose-dependent increase, which became significantly greater than a purely additive effect at a staurosporine concentration of 10 nM. This synergy was maintained at all doses of staurosporine tested. In addition, doses of phorbol 12-myristate 13-acetate (PMA) that do not activate protein kinase C (PKC) (below 10(-9) M) also augmented the zymosan-stimulated release of LTB4. However, doses of PMA above 10(-9) M progressively inhibited the response to levels below that of zymosan alone. Staurosporine at 50 nM completely prevented, and 10(-9) M rhTNF-alpha partially but significantly (P less than 0.02 at 10(-8) M PMA, P less than 0.01 at 10(-7) M PMA) reversed, this high-dose PMA inhibition. PKC activation thus opposes the priming effect of rhTNF-alpha on neutrophils, while PKC inhibition may enhance the ability of rhTNF-alpha to prime PMN for zymosan activation. The combined effect of rhTNF-alpha and staurosporine suggests an intracellular synergy rather than simply a direct action due to increased zymosan receptor expression. Thus there appear to be mechanisms whereby the responses of neutrophils may be augmented without activating PKC. Indeed, kinase activation may even exert a degree of feedback control that is antagonized by rhTNF-alpha treatment.
Petersen
M M
MM
Institute of Nephrology, University of Wales College of Medicine, Cardiff Royal Infirmary, U.K.
Steadman
R
R
Williams
J D
JD
D000470:Alkaloids / Q000276:immunology; D004306:Dose-Response Relationship, Immunologic; D006801:Humans; D007975:Leukotriene B4 / Q000097:blood*; D016177:Macrophage-1 Antigen / Q000276:immunology; D009504:Neutrophils / Q000276:immunology*; D011493:Protein Kinase C / Q000037:antagonists & inhibitors / Q000276:immunology*; D011994:Recombinant Proteins / Q000276:immunology; D019311:Staurosporine; D014409:Tumor Necrosis Factor-alpha / Q000276:immunology*; D015054:Zymosan / Q000276:immunology*
D000470:Alkaloids; D016177:Macrophage-1 Antigen; D011994:Recombinant Proteins; D014409:Tumor Necrosis Factor-alpha; D007975:Leukotriene B4; D015054:Zymosan; D011493:Protein Kinase C; D019311:Staurosporine