P-selectin glycoprotein ligand-1 mediates rolling of human neutrophils on P-selectin.
Neutrophils roll on P-selectin expressed by activated platelets or endothelial cells under the shear stresses in the microcirculation. P-selectin glycoprotein ligand-1 (PSGL-1) is a high affinity ligand for P-selectin on myeloid cells. However, it has not been demonstrated that PSGL-1 contributes to the rolling of neutrophils on P-selectin. We developed two IgG mAbs, PL1 and PL2, that appear to recognize protein-dependent epitopes on human PSGL-1. The mAbs bound to PSGL-1 on all leukocytes as well as on heterologous cells transfected with PSGL-1 cDNA. PL1, but not PL2, blocked binding of 125-I-PSGL-1 to immobilized P-selectin, binding of fluid-phase P-selectin to myeloid and lymphoid leukocytes, adhesion of neutrophils to immobilized P-selectin under static conditions, and rolling of neutrophils on P-selectin-expressing CHO cells under a range of shear stresses. PSGL-1 was localized to microvilli on neutrophils, a topography that may facilitate its adhesive function. These data indicate that (a) PSGL-1 accounts for the high affinity binding sites for P-selectin on leukocytes, and (b) PSGL-1 must interact with P-selectin in order for neutrophils to roll on P-selectin at physiological shear stresses.
Moore
K L
KL
Department of Medicine, University of Oklahoma Health Sciences Center, Oklahoma City 73104-5073.
Patel
K D
KD
Bruehl
R E
RE
Li
F
F
Johnson
D A
DA
Lichenstein
H S
HS
Cummings
R D
RD
Bainton
D F
DF
McEver
R P
RP
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal; D000918:Antibody Specificity; D001483:Base Sequence; D015153:Blotting, Western; D016466:CHO Cells; D002448:Cell Adhesion / Q000502:physiology*; D003001:Cloning, Molecular; D006224:Cricetinae; D017276:Friction; D007962:Leukocytes / Q000276:immunology; D008562:Membrane Glycoproteins / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification / Q000378:metabolism*; D008565:Membrane Proteins / Q000032:analysis; D016253:Microscopy, Immunoelectron; D008871:Microvilli / Q000648:ultrastructure; D008969:Molecular Sequence Data; D009504:Neutrophils / Q000276:immunology / Q000502:physiology* / Q000648:ultrastructure; D019007:P-Selectin; D010980:Platelet Membrane Glycoproteins / Q000378:metabolism*; D011994:Recombinant Proteins; D014162:Transfection
D000911:Antibodies, Monoclonal; D008562:Membrane Glycoproteins; D008565:Membrane Proteins; D019007:P-Selectin; C084882:P-selectin ligand protein; D010980:Platelet Membrane Glycoproteins; D011994:Recombinant Proteins