The N-terminal peptide of PSGL-1 can mediate adhesion to trauma-activated endothelium via P-selectin in vivo

• Published in: Blood 2002-07, Vol.100 (2), p.531-538
• Main Authors: Burch, Erin E., Patil, Vivek R. Shinde, Camphausen, Raymond T., Kiani, Mohammad F., Goetz, Douglas J.
• Format: Article
• Language: English
• Subjects: Animals; Biological and medical sciences; Cardiovascular system; Cell Adhesion; Endothelium, Vascular - injuries; Endothelium, Vascular - metabolism; Endothelium, Vascular - pathology; Humans; Leukocytes - cytology; Leukocytes - physiology; Ligands; Medical sciences; Membrane Glycoproteins - chemistry; Membrane Glycoproteins - metabolism; Membrane Glycoproteins - physiology; Mice; Microspheres; P-Selectin - physiology; Peptide Fragments - chemistry; Peptide Fragments - metabolism; Peptide Fragments - physiology; Pharmacology. Drug treatments; Protein Binding; Recombinant Proteins - chemistry; Recombinant Proteins - metabolism; Vascular wall; Wounds and Injuries
• ISSN: 0006-4971; 1528-0020
• DOI: 10.1182/blood.V100.2.531

P-selectin glycoprotein ligand-1 (PSGL-1) is present on leukocytes and is the major ligand for endothelial expressed P-selectin. A variety of studies strongly suggests that the N-terminal region of PSGL-1 contains the binding site for P-selectin. We hypothesized that this relatively small N-terminal peptide of PSGL-1 is sufficient to support adhesion to P-selectin in vivo. To test this hypothesis, we coated 2 μm–diameter microspheres with a recombinant PSGL-1 construct, termed 19.ek.Fc. The 19.ek.Fc construct consists of the first 19 N-terminal amino acids of mature PSGL-1 linked to an enterokinase cleavage site that, in turn, is linked to human immunoglobulin G Fc. The 19.ek.Fc-coated microspheres were injected into the jugular vein of mice. Intravital microscopy of postcapillary venules within the cremaster muscle of mice revealed that a significantly greater number of 19.ek.Fc microspheres rolled compared with control microspheres. The number of rolling 19.ek.Fc microspheres was significantly diminished by pretreatment of the mice with a monoclonal antibody to P-selectin or by pretreatment of the 19.ek.Fc microspheres with a monoclonal antibody to PSGL-1. Combined, the results indicate that the N-terminal peptide of PSGL-1 can mediate adhesion to trauma-activated microvascular endothelium via P-selectin in vivo.