Stimulation of primed neutrophils by soluble immune complexes: priming leads to enhanced intracellular Ca2+ elevations, activation of phospholipase D, and activation of the NADPH oxidase.
Soluble immune complexes activate a rapid burst of reactive oxidant secretion from neutrophils that have previously been primed with GM-CSF. Binding of these complexes to the cell surface of unprimed neutrophils results in the generation of intracellular Ca2+ transients, but the NADPH oxidase fails to become activated. No phospholipase D activity was observed following the addition of soluble immune complexes to unprimed cells. Upon priming with GM-CSF, the intracellular Ca2+ signal generated following soluble complex binding was greatly extended and phospholipase D was activated: there was also increased phosphorylation of proteins on tyrosine residues and the NADPH oxidase was activated. When Ca2+ influx was prevented, this phospholipase D activity was not observed. This primed oxidase activity was completely inhibited by erbstatin. Treatment of unprimed neutrophils with pervanadate (to inhibit protein tyrosine phosphatases) mimicked the effects of priming in that pervanadate-treated neutrophils secreted reactive oxidants in response to soluble immune complexes. The data indicate that during priming a new signaling pathway is activated that involves Ca2+ influx, phosphorylation on tyrosine residues, phospholipase D activity, and NADPH oxidase activation.
Watson
F
F
School of Biological Sciences, University of Liverpool, United Kingdom.
Edwards
S W
SW
D000814:Aniline Compounds / Q000378:metabolism; D000936:Antigen-Antibody Complex / Q000494:pharmacology*; D002118:Calcium / Q000378:metabolism*; D004789:Enzyme Activation / Q000276:immunology; D016178:Granulocyte-Macrophage Colony-Stimulating Factor / Q000494:pharmacology; D006801:Humans; D006873:Hydroquinones / Q000494:pharmacology; D019255:NADPH Oxidases / Q000378:metabolism*; D009504:Neutrophils / Q000201:enzymology*; D010739:Phospholipase D / Q000378:metabolism; D010766:Phosphorylation; D019000:Phosphotyrosine / Q000032:analysis; D010972:Platelet Activating Factor / Q000031:analogs & derivatives / Q000378:metabolism; D011505:Protein-Tyrosine Kinases / Q000378:metabolism; D017382:Reactive Oxygen Species / Q000378:metabolism; D016897:Respiratory Burst / Q000276:immunology / Q000502:physiology; D015398:Signal Transduction / Q000502:physiology; D014638:Vanadates / Q000494:pharmacology; D014966:Xanthenes / Q000378:metabolism
D000814:Aniline Compounds; D000936:Antigen-Antibody Complex; D006873:Hydroquinones; C029271:O-deacetyl platelet activating factor; D010972:Platelet Activating Factor; D017382:Reactive Oxygen Species; D014966:Xanthenes; C076715:pervanadate; D019000:Phosphotyrosine; C059715:Fluo-3; D014638:Vanadates; D016178:Granulocyte-Macrophage Colony-Stimulating Factor; D019255:NADPH Oxidases; D011505:Protein-Tyrosine Kinases; D010739:Phospholipase D; D002118:Calcium; C048212:erbstatin