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Blood, 15 November 2006, Vol. 108, No. 10, pp. 3352-3359.
Prepublished online as a Blood First Edition Paper on July 18, 2006; DOI 10.1182/blood-2006-04-013912.
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HEMOSTASIS, THROMBOSIS, AND VASCULAR BIOLOGY
Lipid raft adhesion receptors and Syk regulate selectin-dependent rolling under flow conditions
Claire Abbal,
Martine Lambelet,
Debora Bertaggia,
Carole Gerbex,
Manuel Martinez,
Alexandre Arcaro,
Marc Schapira, and
Olivier Spertini
From the Service and Central Laboratory of Hematology, Centre Hospitalier Universitaire Vaudois, Lausanne; and the Department of Pediatrics, Division of Clinical Chemistry and Biochemistry, University Children's Hospital, Zurich, Switzerland.
Selectins and their ligand P-selectin glycoprotein ligand-1 (PSGL-1) mediate leukocyte rolling along inflamed vessels. Cell rolling is modulated by selectin interactions with their ligands and by topographic requirements including L-selectin and PSGL-1 clustering on tips of leukocyte microvilli. Lipid rafts are cell membrane microdomains reported to function as signaling platforms. Here, we show that disruption of leukocyte lipid rafts with cholesterol chelating agents depleted raft-associated PSGL-1 and L-selectin and strongly reduced L-, P-, and E-selectin–dependent rolling. Cholesterol repletion reversed inhibition of cell rolling. Importantly, leukocyte rolling on P-selectin induced the recruitment of spleen tyrosine kinase (Syk), a tyrosine kinase associated to lipid raft PSGL-1. Furthermore, inhibition of Syk activity or expression, with pharmacologic inhibitors or by RNA interference, strongly reduced leukocyte rolling on P-selectin, but not on E-selectin or PSGL-1. These observations identify novel regulatory mechanisms of leukocyte rolling on selectins with a strong dependency on lipid raft integrity and Syk activity.
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