L-selectin ligands expressed by human leukocytes are HECA-452 antibody-defined carbohydrate epitopes preferentially displayed by P-selectin glycoprotein ligand-1

J Immunol. 1999 Nov 1;163(9):5070-8.


Leukocytes express L-selectin ligands critical for leukocyte-leukocyte interactions at sites of inflammation. The predominant leukocyte L-selectin ligand is P-selectin glycoprotein ligand-1 (PSGL-1), which displays appropriate sialyl Lewis x (sLex)-like carbohydrate determinants for L-selectin recognition. Among the sLex-like determinants expressed by human leukocytes is a unique carbohydrate epitope defined by the HECA-452 mAb. The HECA-452 Ag is a critical component of L-selectin ligands expressed by vascular endothelial cells. However, HECA-452 Ag expression on human leukocyte L-selectin ligands has not been assessed. In this study, the HECA-452 mAb blocked 88-99% of neutrophil rolling on, or attachment to, adherent cells expressing L-selectin in multiple experimental systems. A function-blocking anti-PSGL-1 mAb also inhibited L-selectin binding to neutrophils by 89-98%. In addition, the HECA-452 and anti-PSGL-1 mAbs blocked the majority of P-selectin binding to neutrophils. Western blot analysis revealed that PSGL-1 immunoprecipitated from neutrophils displayed HECA-452 mAb-reactive determinants and that PSGL-1 was the predominant scaffold for HECA-452 Ag display. Leukocyte L-selectin ligands also contained sulfated determinants since culturing ligand-bearing cells with NaClO3 abrogated L-selectin binding. Consistent with this, human neutrophils expressed mRNA encoding five different sulfotransferases associated with the generation of selectin ligands: CHST1, CHST2, CHST3, TPST1, and HEC-GlcNAc6ST. Therefore, the HECA-452-defined carbohydrate determinant displayed on PSGL-1 represented the predominant L-selectin and P-selectin ligand expressed by neutrophils.

Publication types

  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Animals
  • Antibodies, Blocking / pharmacology
  • Antibodies, Monoclonal / metabolism*
  • Antibodies, Monoclonal / pharmacology
  • Antigens, Differentiation, T-Lymphocyte
  • Antigens, Neoplasm
  • Blood Circulation / immunology
  • Blood Circulation / physiology
  • COS Cells
  • Carbohydrate Metabolism*
  • Carbohydrates / immunology
  • Cell Movement / immunology
  • Cell Movement / physiology
  • Epitopes / metabolism*
  • Fucosyltransferases / genetics
  • HL-60 Cells
  • Humans
  • L-Selectin / biosynthesis*
  • L-Selectin / metabolism
  • Leukocytes / metabolism*
  • Ligands
  • Membrane Glycoproteins / immunology
  • Membrane Glycoproteins / metabolism*
  • Mice
  • Neutrophils / immunology
  • Neutrophils / metabolism
  • Neutrophils / physiology
  • P-Selectin / immunology
  • P-Selectin / metabolism*
  • Sulfotransferases / biosynthesis
  • Sulfotransferases / genetics
  • Transfection


  • Antibodies, Blocking
  • Antibodies, Monoclonal
  • Antigens, Differentiation, T-Lymphocyte
  • Antigens, Neoplasm
  • CTAGE1 protein, human
  • Carbohydrates
  • Epitopes
  • Ligands
  • Membrane Glycoproteins
  • P-Selectin
  • P-selectin ligand protein
  • L-Selectin
  • Fucosyltransferases
  • galactoside 3-fucosyltransferase
  • Sulfotransferases