Exposure of polymorphonuclear neutrophils (PMNs) to phorbol 12-myristate 13-acetate (PMA) resulted in a concentration-dependent (1–10 ng/ml) inhibition of granule exocytosis induced with the receptor-specific ligands,N-formylmethionyl-leucyl-phenylalanine (FMLP), pepstatin A, 5(S),12(R)-dihydroxy-6, 14-cis-8, 10-trans-eicosatetraenoic acid (LTB4), and acetyl-sn-glyceryl-3-phosphorylcholine (AGEPC), PMA exerted a marginal inhibitory effect on calcium ionophore A23187-induced PMN degranulation, and the PMA analog, 4α-phorbol 12, 13-didecanoate (4α-PDD), was inactive. However, PMA potentiated AGEPC, pepstatin A, FMLP, LTB4, and A23187-stimulated Superoxide anion (O 2 − ) production. The mobilization of intracellular sequestered calcium (Ca2+) by the receptor-specific ligands, as reflected by a rise in the cytosolic-free Ca2+ concentration ([Ca2+]i) in PMNs loaded with the CA2+-sensitive dye, Fura-2, was suppressed by PMA. A protein kinase C (PKC) inhibitor, 1-(5-isoquinolinesulfonyl)-2-methylpiperazine (H-7) reversed the PMA-mediated inhibition of PMN degranulation and intracellular CA2+ mobilization. However, another, but less potent PKC inhibitor,N-(2-guanidinoethyl)-5-isoquinolinesulfonamide (HA 1004), had no effect on the inhibition of PMN activation by PMA.
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Verghese, M. W., C. D. Smith, andR. Snyderman. 1986. Role of guanine nucleotide regulatory protein in polyphosphoinositide degradation and activation of phagocytic leukocytes by chemoattractants.J. Cell. Biochem. 32:59–69.
Snyderman, R., C. D. Smith, andM. W. Verghese. 1986. Model for leukocyte regulation by chemoattractant receptors: Roles of a guanine nucleotide regulatory protein and polyphosphoinositide metabolism.J. Leuk. Biol. 40:785–800.
Berridge, M. J. 1987. Inositol triphosphate and diacylglycerol: Two interacting second messengers.Annu. Rev. Biochem. 56:159–193.
Berridge, M. J., andR. F. Irvine. 1984. Inositol trisphosphate, a novel second messenger in cellular signal transduction.Nature 312:315–321.
Nishizuka, Y. 1984. The role of protein kinase C in cell surface signal transduction and tumor promotion.Nature 308:693–698.
Costagna, M., Y. Takai, K. Kaibuchi, K. Sano, U. Kikkawa, andY. Nishizuka. 1982. Direct activation of calcium-activated, phospholipid-dependent protein kinase by tumor-promoting phorbol esters.J. Biol. Chem. 257:7847–7851.
Nishihira, J., andJ. T. O'Flaherty. 1985. Phorbol myristate acetate receptors in human polymorphonuclear neutrophils.J. Immunol. 135:3439–3447.
Sharkey, N. A., K. L. Leach, andP. M. Blumberg. 1984. Competitive inhibition by diacylglycerol of specific phorbol ester binding.Proc. Natl. Acad. Sci. U.S.A. 81:607–610.
Wolfson, M., L. C. McPhail, V. N. Nasrallah, andR. Snyderman. 1985. Phorbol myristrate acetate mediates redistribution of protein kinase C in human neutrophils. Potential role in the activation of the respiratory burst enzyme.J. Immunol. 135:2057–2062.
O'Flaherty, J. T., andJ. Nishihara. 1987. Arachidonate metabolites, platelet-activating factor, and the mobilization of protein kinase C in human polymorphonuclear neutrophils.J. Immunol. 138:1889–1895.
Smith, R. L., B. J. Bowman, S. S. Iden, G. J. Kolaja, andS. K. Wiser. 1983. Biochemical, metabolic, and morphological characteristics of human neutrophil activation with pepstatin A.Immunology 49:367–377.
Naccache, P. H., T. F. P. Molski, P. Borgeat, J. R. White, andR. I. Sha'afi. 1985. Phorbol esters inhibit the f met-leu-phe and leukotriene B4-stimulated calcium mobilization and enzyme secretion in rabbit neutrophils.J. Biol. Chem. 260:2125–2131.
Zavoico, G. B., S. P. Halenda, R. I. Sha'afi, andM. B. Feinstein. 1985. Phorbol myristate acetate inhibits thrombin-stimulated CA2+ mobilization and phosphatidyl 4,5-bisphosphate hydrolysis in human platelets.Proc. Natl. Acad. Sci. U.S.A. 82:3859–3862.
Okano, Y., H. Takagi, S. Nakashima, T. Tohmatsu, andY. Nozawa. 1985. Inhibitory action of phorbol myristate acetate on histamine secretion and polyphosphoinositide turnover induced by compound 48/80 in mast cells.Biochem. Biophys. Res. Commun. 132:110–117.
Grynkiewicz, G., M. Poenie, andR. Y. Tsien. 1985. A new generation of Ca2+-indicators with greatly improved fluorescence properties.J. Biol. Chem. 260:3440–3450.
Henson, P. M., B. Zanolari, N. A. Schwartzman, andS. R. Hong. 1978. Intracellular control of human neutrophil secretion: 1. C5a-induced stimulus-specific desensitization and the effects of cytochalasin B.J. Immunol. 131:851–855.
Bergmeyer, H. U., E. Bernt, andB. Hess. 1965. Lactic dehydrogense.In Methods of Enzymatic Analysis. H. U. Burgmeyer, editor. Verlag Chemie, Academic Press, New York. 736–743.
Smith, R. J., andS. S. Iden. 1981. Modulation of human neutrophil Superoxide anion generation by the calcium antagonist 8-(N,N-diethylamino)-octyl-(3,4,5-trimethoxy)benzoate hydrochloride.J. Reticuloendothel Soc. 29:215–225.
Sha'afi, R. I., J. R. White, T. F. P. Molski, J. Shefcyk, M. Volpi, P. H. Naccache, andM. B. Feinstein. 1983. Phorbol 12-myristate 13-acetate activates rabbit neutrophils without an apparent rise in the level of the intracellular free calcium.Biochem. Biophys. Res. Commun. 114:638–645.
Korchak, H. M., K. Vienne, L. E. Rutherford, C. Wilkenfeld, M. C. Finkelstein, andG. Weissmann. 1984.J. Biol. Chem. 259:4076–4082.
Verghese, M. W., L. Charles, L. Jakoi, S. B. Dillon, andR. Snyderman. 1987. Role of a guanine nucleotide regulatory protein in the activation of phospholipase C by different chemoattractants.J. Immunol. 138:4374–4380.
Okjima, F., andM. Ui. 1984. ADP-ribosylation of the specific membrane protein by isletactivating protein, pertussis toxin, associated with inhibition of a chemotactic peptide-induced arachidonate release in neutrophils.J. Biol. Chem. 259:13863–13871.
Bokoch, G. M., andA. G. Gilman. 1984. Inhibition of receptor-mediated release of arachidonate acid by pertussis toxin.Cell 39:301–308.
Bradford, P. G., andR. P. Rubin. 1985. Characterization of formylmethionylleucylphenylalanine stimulation of inositol trisphosphate accumulation in rabbit neutrophils.Mol. Pharmacol. 27:74–78.
Kawamoto, S., andH. Hidaka. 1984. 1-(5-isoquinolinesulfonyl)-2-methylpiperazine (H-7) is a selective inhibitor of protein kinase C in rabbit platelets.Biochem. Biophys. Res. Commun. 125:258–264.
Hidaka, H., M. Inagaki, S. Kawamoto, andY. Sassaki. 1984. Isoquinolinesulfonamides, novel and potent inhibitors of cyclic nucleotide protein kinase and protein kinase C.Biochemistry 23:5036–5041.
Sha'afi, R. I., T. F. P. Molski, C. K. Huang, andP. H. Naccache. 1986. The inhibition of neutrophil responsiveness caused by phorbol esters is blocked by the protein kinase C inhibitor H-7.Biochem. Biophys. Res. Commun. 137:50–60.
White, J. R., C. K. Huang, J. R. Hill, P. H. Naccache, E. L. Becker, andR. I. Sha'afi. 1984. Effect of phorbol 12-myristate 13-acetate and its analog 4α-phorbol 12,13-didecanoate on protein phosphorylation and lysosomal enzyme release in rabbit neutrophils.J. Biol. Chem. 259:8605–8611.
Katada, T., A. G. Gilman, Y. Watanabe, S. Bauer, andK. H. Jakobs. 1985. Protein kinase C phosphorylates the inhibitory guanine-nucleotide regulatory component and apparently suppresses its function in hormonal inhibition of adenylate cyclase.Eur. J. Biochem. 151:431–437.
Kikuchi, A. O. Kozawa, Y. Hamamori, K. Kaibuchi, andY. Takai. 1986. Inhibition of chemotactic peptide-induced phosphoinositide hydrolysis by phorbol esters through the activation of protein kinase C in differentiated leukemia (HL-60) cells.Cancer Res. 46:3401–3406.
Kikuchi, A., K. Ikeda, O. Kozawa, andY. Takai. 1987. Modes of inhibitory action of protein kinase C in the chemotactic peptide-induced formation of inositol phosphates in differentiated leukemic (HL-60) cells.J. Biol. Chem. 262:6766–6770.
Smith, C. D., R. J. Uhring, andR. Snyderman. 1987. Nucelotide regulatory proteinmediated activation of phospholipase C in human polymorphonuclear leukocytes is disrupted by phorbol ester.J. Biol. Chem. 262:6121–6127.
Watson, S. P., andE. G. Lapetina. 1985. 1,2-Diacyglycerol and phorbol ester inhibit agonist-induced formation of inositol phosphates in human platelets: Possible implications for negative feedback regulation of inositol phospholipid hydrolysis.Proc. Natl. Acad. Sci. U.S.A. 82:2623–2626.
Brown, K. D., D. M. Blakeley, M. H. Hamon, M. S. Laurie, andA. N. Corps. 1987. Protein kinase C-mediated negative-feedback inhibition of unstimulated and bombesin-stimulated polyphosphoinositide hydrolysis in swiss-mouse 3T3 cells.Biochem. J. 245:631–639.
Wilson, J. L., andL. C. McPhail 1987. Altered activation parameters of protein kinase C in stimulated human neutrophils.Fed. Proc. 46:1033.
Wright, C. D., andM. D. Hoffman. 1986. The protein kinase C inhibitors H-7 and H-9 fail to inhibit human neutrophil activation.Biochem. Biophys. Res. Commun. 135:749–755.
Umezawa, H., T. Aoyagi, H. Morishima, M. Matsuzaki, M. Hamada, andT. Takeuchi. 1970. Pepstatin, a new pepsin inhibitor produced by actinomycetes.J. Antibiot. 23:259–262.
Ackerman, S. K., andS. D. Douglas. 1979. Pepstatin A—a human leukocyte chemoattractant.Clin. Immunol. Immunopathol. 14:244–250.
Smith, R. J., B. J. Bowman, andS. S. Iden. 1982. Pepstatin A-induced degranulation and Superoxide anion generation by human neutrophils.Clin. Immunol. Immunopathol. 22:83–93.
Smith, R. J., B. J. Bowman, S. S. Iden, G. J. Kolaia, andS. K. Wiser. 1983. Biochemical metabolic and morphological characteristics of human neutrophil activation with pepstatin A.Immunology 49:367–377.
Nelson, R.D., S. M. Ackerman, V. D. Fiegel, M. P. Bowman, andS. D. Douglas. 1979. Cytotaxin receptors of neutrophils: Evidence that F-methionyl peptides and pepstatin share a common receptor.Infect. Immun. 26:996–999.
Gerard, C., L. C. McPhail, A. Marfat, N. Stimler-Gerard, D. A. Bass, andC. E. McCall 1986. Role of protein kinase in stimulation of human polymorphonuclear leukocyte oxidative metabolism by various agonists: Differential effects of a novel protein kinase inhibitor.J. Clin. Invest. 77:61–65.
Pike, M. C., L. Jakoi, L. C. McPhail, andR. Snyderman. 1986. Chemoattractant-mediated stimulation of the respiratory burst in human polymorphonuclear leukocytes may require appearance of protein kinase activity in the cells particulate fraction.Blood 67:909–913.
Della Bianca, V., M. Grzeskowiak, M. A. Cassatella, L. Zeni, andF. Rossi. 1986. Phorbol 12-myristate 13-acetate potentiates the respiratory burst while inhibits phosphoinositide hydrolysis and calcium mobilization by formyl-methionyl-leucyl-phenylalanine in human neutrophils.Biochem. Biophys. Res. Commun. 135:556–565.
Dewald, B., T. G. Payne, andM. Baggiolini. 1984. Activation of NADPH oxidase of human neutrophils. Potentiation of chemotactic peptide by a diacylglycerol.Biochem. Biophys. Res. Commun. 125:367–373.
Salari, H., P. Braquet, P. Naccache, andP. Borgeat. 1985. Characterization of effect ofN-formyl-methionyl-leucyl-phenylalanine on leukotriene syntheses in human polymorphonuclear leukocytes.Inflammation 9:127–138.
Sisson, J. H., S. M. Prescott, T. M. McIntyre, andG. A. Zimmerman. 1987. Production of platelet-activating factor by stimulated human polymorphonuclear leukocytes: Correlation of synthesis with release, functional events, and leukotriene B4 metabolism.J. Immunol. 138:3918–3926.
Dewald, B., andM. Baggiolini. 1985. Activation of NADPH oxidase in human neutrophils. Synergism between FMLP and the neutrophil products PAF and LTB4.Biochem. Biophys. Res. Commun. 128:297–304.
Baggiolini, M. andB. Dewald. 1986. Stimulus amplification by PAF and LTB4 in human neutrophils.Pharmacol. Res. Commun. 18(Suppl):51–59.
DiVirgilio, F., D. P. Lew, andT. Pozzan. 1984. Protein kinase C activation of physiological processes in human neutrophils at vanishingly small cytosolic Ca2+ levels.Nature 310:691–693.
Penfield, A., andM. M. Dale. 1984. Synergism between A23187 and 1-oleoyl-2-acetylglycerol in superoxide production by human neutrophils.Biochem. Biophys. Res. Commun. 125:332–336.
Volpi, M., T. F. P. Molski, P. H. Naccache, M. B. Feinstein, andR. I. Sha'afi. 1985. Phorbol 12-myristate 13-acetate potentiates the action of the calcium ionophore in stimulating arachidonic acid release and production of phosphatidic acid in rabbit neutrophils.Biochem. Biophys. Res. Commun. 128:594–606.
Liles, W. C., K. E. Meier, andW. R. Henderson. 1987. Phorbol myristate acetate and the calcium ionophore A23187 synergistically induce release of LTB4 by human neutrophils: Involvement of protein kinase C in regulation of the 5-lipoxygenase pathway.J. Immunol. 138:3396–3402.
Berkow, R. L., R. W. Dodson, andA. S. Kraft. 1987. Th effect of a protein kinase C inhibitor, H-7, on human neutrophil oxidative burst and degranulation.J. Leuk. Biol. 41:441–446.
Harwath, L., C. E. McCall, D. A. Bass, andL. C. McPhail 1987. Inhibition of human neutrophil chemotaxis by the protein kinase inhibitor, 1-(5-isoquinolinesulfonyl)piperazine.J. Immunol. 139:3055–3061.
Salzer, W., C. Gerard, andC. McCall 1987. Effect of an inhibitor of protein kinase C on human polymorphonuclear leukocyte degranulation.Biochem. Biophys. Res. Commun. 148:747–754.
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Smith, R.J., Justen, J.M. & Sam, L.M. Function and stimulus-specific effects of phorbol 12-myristate 13-acetate on human polymorphonuclear neutrophils: Autoregulatory role for protein kinase C in signal transduction. Inflammation 12, 597–611 (1988). https://doi.org/10.1007/BF00914321
- Calcium Ionophore
- Polymorphonuclear Neutrophil
- Granule Exocytosis