title: Modulation of TNF-alpha-priming and stimulation-dependent superoxide generation in human neutrophils by protein kinase inhibitors.
abstract: Human peripheral blood polymorphonuclear leukocytes (HPPMN) from healthy individuals are not primed and, hence, weak stimulation-dependent responses are induced by certain stimuli which bind to membrane receptors. When HPPMN were exposed to recombinant human tumor necrosis factor alpha (rHuTNF-alpha) or recombinant human granulocyte colony stimulating factor (rG-CSF), they underwent priming and the rate of superoxide anion (O.-2) generation was increased by subsequent exposure to formyl-methionyl-leucyl-phenylalanine (FMLP) or opsonized zymosan (OZ). However, the degree of enhancement was very small upon exposure to phorbol myristate acetate (PMA) or dioctanoyl glycerol (DOG). The oxygen burst induced by FMLP or OZ was inhibited by genistein and alpha-cyano-3-ethoxy-4-hydroxy-5-phenylthiomethylcinnamamid (ST638), which are inhibitors of tyrosine kinase (TK), and was enhanced by 1-(5-isoquinoline-sulfonyl)-3-methyl-piperazine (H-7) and staurosporine, which are inhibitors of protein kinase C (PKC). Without priming, however, O.-2 generation from HPPMN by high concentrations of FMLP was not inhibited strongly by genistein or ST638. On the contrary, the oxygen burst induced by PMA or DOG was stimulated by genistein or ST638 and was inhibited by H-7 or staurosporine. Furthermore, O.-2 generation by guinea pig peritoneal neutrophils, which are already primed in vivo, was induced markedly by FMLP by a mechanism which was stimulated by a low concentration of genistein or ST638. Thus, FMLP-mediated O.-2-generation of HPPMN is coupled with rHuTNF-alpha- or rG-CSF-priming and is inhibited by TK inhibitors, whereas PMA- or DOG-induced O.-2 generation is not coupled with TNF-alpha or G-CSF-priming and is inhibited by PKC inhibitors. These results suggest that both PKC and TK play critical roles in the regulatory mechanism of priming and NADPH-oxidase activation in neutrophils.
authors.affiliation: Department of Tumor Biology, MD Anderson Cancer Center, University of Texas, Houston 77030.
authors.forename: T
authors.initials: T
authors.lastname: Utsumi
authors.forename: J
authors.initials: J
authors.lastname: Klostergaard
authors.forename: K
authors.initials: K
authors.lastname: Akimaru
authors.forename: K
authors.initials: K
authors.lastname: Edashige
authors.forename: E F
authors.initials: EF
authors.lastname: Sato
authors.forename: K
authors.initials: K
authors.lastname: Utsumi
mesh_terms: D019307:1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine; D000470:Alkaloids / Q000494:pharmacology; D000818:Animals; D002934:Cinnamates / Q000494:pharmacology; D004075:Diglycerides / Q000494:pharmacology; D019833:Genistein; D006168:Guinea Pigs; D006801:Humans; D007529:Isoflavones / Q000494:pharmacology; D007546:Isoquinolines / Q000494:pharmacology; D009240:N-Formylmethionine Leucyl-Phenylalanine / Q000494:pharmacology; D009504:Neutrophils / Q000187:drug effects / Q000378:metabolism*; D009895:Opsonin Proteins; D010879:Piperazines / Q000494:pharmacology; D047428:Protein Kinase Inhibitors; D011505:Protein-Tyrosine Kinases / Q000037:antagonists & inhibitors; D011994:Recombinant Proteins / Q000494:pharmacology; D019311:Staurosporine; D013440:Sulfides / Q000494:pharmacology; D013481:Superoxides / Q000097:blood*; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology*; D015054:Zymosan / Q000494:pharmacology
chemical_list: D000470:Alkaloids; D002934:Cinnamates; D004075:Diglycerides; D007529:Isoflavones; D007546:Isoquinolines; D009895:Opsonin Proteins; D010879:Piperazines; D047428:Protein Kinase Inhibitors; D011994:Recombinant Proteins; D013440:Sulfides; D014409:Tumor Necrosis Factor-alpha; C044295:1,2-dioctanoylglycerol; C060747:ST 638; D013481:Superoxides; D009240:N-Formylmethionine Leucyl-Phenylalanine; D019307:1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine; D015054:Zymosan; D019833:Genistein; D011505:Protein-Tyrosine Kinases; D019311:Staurosporine; D013755:Tetradecanoylphorbol Acetate