E-selectin receptors on human leukocytes.
Selectins on activated vascular endothelium mediate inflammation by binding to complementary carbohydrates on circulating neutrophils. The human neutrophil receptor for E-selectin has not been established. We report here that sialylated glycosphingolipids with 5 N-acetyllactosamine (LacNAc, Galbeta1-4GlcNAcbeta1-3) repeats and 2 to 3 fucose residues are major functional E-selectin receptors on human neutrophils. Glycolipids were extracted from 10(10) normal peripheral blood human neutrophils. Individual glycolipid species were resolved by chromatography, adsorbed as model membrane monolayers and selectin-mediated cell tethering and rolling under fluid shear was quantified as a function of glycolipid density. E-selectin-expressing cells tethered and rolled on selected glycolipids, whereas P-selectin-expressing cells failed to interact. Quantitatively minor terminally sialylated glycosphingolipids with 5 to 6 LacNAc repeats and 2 to 3 fucose residues were highly potent E-selectin receptors, constituting more than 60% of the E-selectin-binding activity in the extract. These glycolipids are expressed on human blood neutrophils at densities exceeding those required to support E-selectin-mediated tethering and rolling. Blocking glycosphingolipid biosynthesis in cultured human neutrophils diminished E-selectin, but not P-selectin, adhesion. The data support the conclusion that on human neutrophils the glycosphingolipid NeuAcalpha2-3Galbeta1-4GlcNAcbeta1-3[Galbeta1-4(Fucalpha1-3)GlcNAcbeta1-3](2)[Galbeta1-4GlcNAcbeta1-3](2)Galbeta1-4GlcbetaCer (and closely related structures) are functional E-selectin receptors.
Nimrichter
Leonardo
L
Department of Pharmacology and Molecular Sciences, The Johns Hopkins School of Medicine, Baltimore, MD 21205, USA.
Burdick
Monica M
MM
Aoki
Kazuhiro
K
Laroy
Wouter
W
Fierro
Mark A
MA
Hudson
Sherry A
SA
Von Seggern
Christopher E
CE
Cotter
Robert J
RJ
Bochner
Bruce S
BS
Tiemeyer
Michael
M
Konstantopoulos
Konstantinos
K
Schnaar
Ronald L
RL
D000606:Amino Sugars / Q000737:chemistry; D002240:Carbohydrate Sequence; D002448:Cell Adhesion; D019040:E-Selectin / Q000097:blood*; D005643:Fucose / Q000737:chemistry; D005732:Gangliosides / Q000097:blood* / Q000737:chemistry / Q000302:isolation & purification; D006801:Humans; D066298:In Vitro Techniques; D036904:Leukocyte Rolling; D007962:Leukocytes / Q000378:metabolism*; D008969:Molecular Sequence Data; D015394:Molecular Structure; D009504:Neutrophils / Q000378:metabolism; D021241:Spectrometry, Mass, Electrospray Ionization; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
D000606:Amino Sugars; D019040:E-Selectin; D005732:Gangliosides; D005643:Fucose; C000458:N-acetyllactosamine