Discordant expression of selectin ligands and sialyl Lewis x-related epitopes on murine myeloid cells

Blood. 2002 Dec 15;100(13):4485-94. doi: 10.1182/blood-2002-06-1799. Epub 2002 Aug 29.


Murine leukocytes are thought to express alpha2-3-sialylated and alpha1-3-fucosylated selectin ligands such as sialyl Lewis x (sLe(x)), although monoclonal antibodies (mAbs) to sLe(x) or Le(x) reportedly do not bind to murine leukocytes. We observed that P- and E-selectin bound to pronase-sensitive ligands on murine monocytic WEHI-3 cells and murine neutrophils, indicating that the ligands for both selectins are glycoproteins. CSLEX-1, HECA-452, and other widely used mAbs to sLe(x) and Le(x) did not bind to WEHI-3 cells and bound at very low levels to murine neutrophils. Only the anti-sLe(x) mAbs 2H5 and KM93, which also recognize nonfucosylated glycans, bound to WEHI-3 cells. 2H5 and KM93 bound to pronase-resistant structures, indicating that the mAbs did not identify selectin ligands. Treatment of WEHI-3 cells with glycosidases or chlorate demonstrated that sialic acid modifications, alpha1-3-galactosylation, or sulfation did not mask epitopes for mAbs to sLe(x) or Le(x). Compared to human promyelocytic HL-60 cells, WEHI-3 cells and murine neutrophils expressed low alpha1-3-fucosyltransferase activities. Consistent with very low endogenous fucosylation, forced fucosylation of intact WEHI-3 cells or murine neutrophils by exogenous alpha1-3-fucosyltransferase FTVI and GDP-fucose created many new epitopes for anti-sLe(x) mAbs such as HECA-452 and CSLEX-1. Nevertheless, forced fucosylation of intact cells did not significantly augment their ability to bind to fluid-phase P- or E-selectin or to roll on immobilized P- or E-selectin under flow. These data suggest that murine myeloid leukocytes fucosylate only a few specific glycans, which interact preferentially with P- and E-selectin.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Animals
  • Antibodies, Monoclonal / immunology
  • Cell Line
  • E-Selectin / metabolism*
  • Epitopes / metabolism*
  • Fucose / immunology
  • Fucosyltransferases / biosynthesis
  • Fucosyltransferases / pharmacology
  • Galactose / immunology
  • HL-60 Cells / metabolism
  • Humans
  • Ligands
  • Mice
  • Myeloid Cells / immunology
  • Myeloid Cells / metabolism*
  • Neuraminidase / pharmacology
  • Neutrophils / metabolism
  • Oligosaccharides / immunology
  • Oligosaccharides / metabolism*
  • P-Selectin / metabolism*
  • Polysaccharides / metabolism
  • Pronase / pharmacology
  • Protein Binding
  • Sialyl Lewis X Antigen
  • Species Specificity
  • Substrate Specificity
  • Sulfates / immunology
  • alpha-Galactosidase / pharmacology


  • Antibodies, Monoclonal
  • E-Selectin
  • Epitopes
  • Ligands
  • Oligosaccharides
  • P-Selectin
  • Polysaccharides
  • Sialyl Lewis X Antigen
  • Sulfates
  • Fucose
  • Fucosyltransferases
  • galactoside 3-fucosyltransferase
  • Neuraminidase
  • alpha-Galactosidase
  • Pronase
  • Galactose