Cell volume-dependent regulation of L-selectin shedding in neutrophils. A role for p38 mitogen-activated protein kinase.
Neutrophil-mediated organ damage is a common feature of many disease states. We previously demonstrated that resuscitation with hypertonic salt solutions prevented the endotoxin-induced leukosequestration and consequent lung injury, and this effect was partially attributed to an altered surface expression of adhesion molecules, CD11b and L-selectin. In this study we investigated the mechanisms whereby osmotic stress evokes L-selectin shedding. The metalloprotease inhibitor RO 31-9790 prevented the osmotic down-regulation of L-selectin, indicating that this process was catalyzed by the same "sheddase" responsible for L-selectin cleavage induced by diverse inflammatory stimuli. The trigger for hypertonic shedding was cell shrinkage and not increased osmolarity, ionic strength, or intracellular pH. Volume reduction caused robust tyrosine phosphorylation and its inhibition by genistein and erbstatin abrogated shedding. Shrinkage stimulated tyrosine kinases Hck, Syk, and Pyk2, but prevention of their activation by the Src-family inhibitor PP1 failed to affect the L-selectin response. Hypertonicity elicited the Src family-independent activation of p38, and the inhibition of this kinase by SB203580 strongly reduced shedding. p38 was also essential for the N-formyl-methionyl-leucyl-phenylalanine- and lipopolysaccharide-induced shedding but not the phorbol ester-induced shedding. Thus, cell volume regulates L-selectin surface expression in a p38-mediated, metalloprotease-dependent manner. Moreover, p38 has a central role in shedding induced by many inflammatory mediators.
Department of Surgery, Toronto General Hospital and the University of Toronto, Toronto, Ontario M5G 2C4, Canada.
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Rizoli
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Rotstein
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Kapus
D017871:Calcium-Calmodulin-Dependent Protein Kinases / Q000097:blood*; D048429:Cell Size / Q000187:drug effects; D003600:Cytosol / Q000502:physiology; D004791:Enzyme Inhibitors / Q000494:pharmacology; D019833:Genistein / Q000494:pharmacology; D006801:Humans; D006863:Hydrogen-Ion Concentration; D006877:Hydroxamic Acids; D006982:Hypertonic Solutions; D007093:Imidazoles / Q000494:pharmacology; D066298:In Vitro Techniques; D019041:L-Selectin / Q000097:blood* / Q000235:genetics; D016177:Macrophage-1 Antigen / Q000097:blood; D008666:Metalloendopeptidases / Q000037:antagonists & inhibitors; D020928:Mitogen-Activated Protein Kinases; D009504:Neutrophils / Q000166:cytology* / Q000187:drug effects / Q000502:physiology*; D019000:Phosphotyrosine / Q000097:blood; D011480:Protease Inhibitors / Q000494:pharmacology; D011725:Pyridines / Q000494:pharmacology; D048051:p38 Mitogen-Activated Protein Kinases
D004791:Enzyme Inhibitors; D006877:Hydroxamic Acids; D006982:Hypertonic Solutions; D007093:Imidazoles; D016177:Macrophage-1 Antigen; D011480:Protease Inhibitors; D011725:Pyridines; C097661:Ro 31-9790; D019041:L-Selectin; D019000:Phosphotyrosine; D019833:Genistein; D017871:Calcium-Calmodulin-Dependent Protein Kinases; D020928:Mitogen-Activated Protein Kinases; D048051:p38 Mitogen-Activated Protein Kinases; D008666:Metalloendopeptidases; C093642:SB 203580