Glycolipids support E-selectin-specific strong cell tethering under flow.
This study provides functional evidence that glycosphingolipids constitute ligands for E-selectin but not P-selectin. Chinese hamster ovary (CHO) cells expressing E-selectin (CHO-E) or P-selectin (CHO-P) were perfused over alpha2,3-sialyl Lewis X (alpha2,3-sLe(x)) presented as the hexaosylceramide glycosphingolipid adsorbed in a monolayer containing phosphatidylcholine and cholesterol. CHO-E cells tethered extensively and formed slow, stable rolling interactions with alpha2,3-sLe(x) glycosphingolipid but not with the comparable alpha2,6-sLe(x) glycosphingolipid. Tethering/rolling varied with wall shear stress, selectin density, and ligand density. In contrast, alpha2,3-sLe(x) glycosphingolipid supported only limited, fast CHO-P cell rolling. As calculated from a stochastic model of cell rolling, the step size between successive bond releases from the alpha2,3-sLe(x) glycosphingolipid was smaller for CHO-E than CHO-P cells, whereas the opposite effect was observed for the waiting time between these events. Detachment assays revealed stronger adhesive interactions of CHO-E than CHO-P cells with alpha2,3-sLe(x) glycosphingolipid. These findings indicate that glycosphingolipids expressing an appropriate oligosaccharide mediate cell tethering/rolling via E-selectin but not P-selectin.
Burdick
M M
MM
Department of Chemical Engineering, Johns Hopkins University, Baltimore, Maryland 21218, USA.
Bochner
B S
BS
Collins
B E
BE
Schnaar
R L
RL
Konstantopoulos
K
K
D000818:Animals; D016466:CHO Cells; D019556:COS Cells; D002448:Cell Adhesion / Q000187:drug effects / Q000502:physiology; D002457:Cell Extracts / Q000494:pharmacology; D002460:Cell Line; D002465:Cell Movement / Q000187:drug effects / Q000502:physiology*; D006224:Cricetinae; D004305:Dose-Response Relationship, Drug; D019040:E-Selectin / Q000235:genetics / Q000378:metabolism*; D000939:Epitopes / Q000378:metabolism; D006028:Glycosphingolipids / Q000378:metabolism* / Q000494:pharmacology; D006801:Humans; D016256:Lewis X Antigen / Q000378:metabolism; D008024:Ligands; D008055:Lipids / Q000302:isolation & purification / Q000494:pharmacology; D008954:Models, Biological; D009504:Neutrophils / Q000737:chemistry / Q000187:drug effects / Q000378:metabolism*; D019007:P-Selectin / Q000235:genetics / Q000378:metabolism; D013269:Stochastic Processes; D013314:Stress, Mechanical; D014162:Transfection
D002457:Cell Extracts; D019040:E-Selectin; D000939:Epitopes; D006028:Glycosphingolipids; D016256:Lewis X Antigen; D008024:Ligands; D008055:Lipids; D019007:P-Selectin