Phospholipase D-derived phosphatidic acid is involved in the activation of the CD11b/CD18 integrin in human eosinophils.
Priming of human eosinophils is an essential event for the respiratory burst induced by serum-opsonized particles [serum-treated zymosan (STZ)]. In this study we have found that treatment of eosinophils with platelet-activating factor (PAF) leads to activation of phospholipase D. Inhibition of the formation of phospholipase D-derived products by ethanol resulted in about 90% inhibition of PAF-induced binding of fluorescent STZ particles to the cells, but only when ethanol was added to the cells before treatment with PAF. When ethanol was added after treatment with PAF, only a minor inhibition of the STZ binding and STZ-induced response was observed. These results indicate that phospholipase D-derived phosphatidic acid is involved in PAF priming, without having an effect on STZ stimulation. In the presence of propranolol, which inhibits phosphatidic acid-phosphatase activity, binding of STZ particles to human eosinophils induced by suboptimal concentrations of PAF was enhanced, indicating that phosphatidic acid and not diradylglyceride is the relevant molecule derived from phospholipase D activity. Addition of cell-permeant diC8-phosphatidic acid (DiC8-PA) to human eosinophils resulted in CD11b/CD18-dependent adhesion, both to STZ particles and fibronectin-coated wells, without significant upregulation of CD11b/CD18. The DiC8-PA-induced adhesion was not mediated via the fatty acid moiety, because other C8-lipids such as 1,2-diC8-phosphatidylcholine, 1-C8-monoacylglycerol or C8-ceramide were without effect. Activation of protein kinase C with PMA or 1,2-diC8-diacylglycerol did result in enhanced STZ binding. However, under these latter conditions upregulation of CD11b/CD18 was observed. Taken together, these results suggest that phospholipase D-derived PA is involved in changing the affinity of the CD11b/CD18 integrin for its ligands.
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Central Laboratory of the Netherlands Red Cross Blood Transfusion Service and Laboratory for Experimental and Clinical Immunology, Academic Medical Center, University of Amsterdam, Plesmanlaan 125, 1066 CX Amsterdam, The Netherlands. a_tool@clb.nl
Blom
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Roos
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Verhoeven
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D018821:CD18 Antigens / Q000378:metabolism*; D002118:Calcium / Q000378:metabolism; D002448:Cell Adhesion / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D004075:Diglycerides / Q000494:pharmacology; D004789:Enzyme Activation / Q000187:drug effects; D004804:Eosinophils / Q000166:cytology / Q000187:drug effects / Q000276:immunology / Q000378:metabolism*; D000431:Ethanol / Q000494:pharmacology; D005353:Fibronectins / Q000378:metabolism; D006801:Humans; D016177:Macrophage-1 Antigen / Q000378:metabolism*; D010712:Phosphatidic Acids / Q000378:metabolism / Q000494:pharmacology / Q000502:physiology*; D010739:Phospholipase D / Q000037:antagonists & inhibitors / Q000378:metabolism*; D010972:Platelet Activating Factor / Q000494:pharmacology; D011433:Propranolol / Q000494:pharmacology; D011485:Protein Binding / Q000187:drug effects; D016897:Respiratory Burst / Q000187:drug effects; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D015854:Up-Regulation / Q000187:drug effects; D015054:Zymosan / Q000378:metabolism / Q000494:pharmacology
D018821:CD18 Antigens; D004075:Diglycerides; D005353:Fibronectins; D016177:Macrophage-1 Antigen; D010712:Phosphatidic Acids; D010972:Platelet Activating Factor; C084527:dioctanoylphosphatidic acid; C044295:1,2-dioctanoylglycerol; D000431:Ethanol; D015054:Zymosan; D011433:Propranolol; D010739:Phospholipase D; D013755:Tetradecanoylphorbol Acetate; D002118:Calcium