Diverging signal transduction pathways activated by interleukin-8 and related chemokines in human neutrophils: interleukin-8, but not NAP-2 or GRO alpha, stimulates phospholipase D activity.
Interleukin-8 (IL-8) and the structurally related cytokines neutrophil-activating peptide-2 (NAP-2) and GRO alpha are powerful chemotactic agents for human neutrophils. Although these three chemokines act by binding to overlapping but not identical receptor subsets, the data available to date have stressed the similarities in their mechanisms of action. The present studies were undertaken to further our understanding of the signal transduction mechanisms associated with these neutrophil agonists. IL-8, NAP-2, and GRO alpha stimulated similar increases in the level of cytoplasmic free calcium. They were also shown to stimulate qualitatively similar increases in the levels of protein tyrosine phosphorylation. In contrast, only IL-8 enhanced the formation of phosphatidylethanol (PEt), the product catalyzed by phospholipase D (PLD) in the presence of ethanol. The formation of PEt stimulated by IL-8 was inhibited by pertussis toxin and the tyrosine kinase inhibitors erbstatin and herbimycin A. The ability of IL-8 to stimulate the activity of PLD was additively enhanced, or primed, by cytochalasin B and by tumor necrosis factor alpha. Although all three chemokines increased the level of free cytoplasmic calcium to the same extent, IL-8 was significantly more potent than either NAP-2 or GRO alpha with respect to its ability to enhance CD11b expression and to stimulate chemotactic and oxidative responses. The differences between IL-8, NAP-2, and GRO alpha in their ability to stimulate PLD is likely to be related to their respective binding affinities for the two IL-8 receptors (IL-8R-A and IL-8R-B). These results suggest that the signalling pathways activated by IL-8R-A and IL-8R-B diverge at a step preceding the activation of PLD.
L'Heureux
G P
GP
Centre de Recherche en Rhumatologie et Immunologie, Centre de Recherche du CHUL, Ste-Foy, Québec, Canada.
Bourgoin
S
S
Jean
N
N
McColl
S R
SR
Naccache
P H
PH
D016227:Benzoquinones; D002118:Calcium / Q000378:metabolism; D054360:Chemokine CXCL1; D019743:Chemokines, CXC; D002630:Chemotactic Factors / Q000494:pharmacology*; D003571:Cytochalasin B / Q000494:pharmacology; D016207:Cytokines / Q000494:pharmacology; D004789:Enzyme Activation / Q000187:drug effects; D006133:Growth Substances / Q000494:pharmacology*; D006801:Humans; D006873:Hydroquinones / Q000494:pharmacology; D036341:Intercellular Signaling Peptides and Proteins; D016209:Interleukin-8 / Q000494:pharmacology*; D047029:Lactams, Macrocyclic; D016177:Macrophage-1 Antigen / Q000096:biosynthesis / Q000235:genetics; D018375:Neutrophil Activation / Q000187:drug effects*; D009504:Neutrophils / Q000187:drug effects* / Q000201:enzymology / Q000502:physiology; D010455:Peptides / Q000494:pharmacology*; D037342:Pertussis Toxin; D010739:Phospholipase D / Q000378:metabolism*; D010766:Phosphorylation / Q000187:drug effects; D011499:Protein Processing, Post-Translational / Q000187:drug effects; D011505:Protein-Tyrosine Kinases / Q000378:metabolism; D011809:Quinones / Q000494:pharmacology; D018123:Receptors, Interleukin / Q000145:classification / Q000187:drug effects / Q000378:metabolism; D023062:Receptors, Interleukin-8A; D016897:Respiratory Burst / Q000187:drug effects; D017828:Rifabutin / Q000031:analogs & derivatives; D015398:Signal Transduction / Q000187:drug effects*; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology; D010566:Virulence Factors, Bordetella / Q000494:pharmacology; D001620:beta-Thromboglobulin
D016227:Benzoquinones; C475829:CXCL1 protein, human; D054360:Chemokine CXCL1; D019743:Chemokines, CXC; D002630:Chemotactic Factors; D016207:Cytokines; D006133:Growth Substances; D006873:Hydroquinones; D036341:Intercellular Signaling Peptides and Proteins; D016209:Interleukin-8; D047029:Lactams, Macrocyclic; D016177:Macrophage-1 Antigen; C061615:PPBP protein, human; D010455:Peptides; D011809:Quinones; D018123:Receptors, Interleukin; D023062:Receptors, Interleukin-8A; D014409:Tumor Necrosis Factor-alpha; D010566:Virulence Factors, Bordetella; D001620:beta-Thromboglobulin; D017828:Rifabutin; D003571:Cytochalasin B; C009862:connective tissue-activating peptide; C020754:herbimycin; D037342:Pertussis Toxin; D011505:Protein-Tyrosine Kinases; D010739:Phospholipase D; D002118:Calcium; C048212:erbstatin