Functional characterization of L-selectin ligands on human neutrophils and leukemia cell lines: Evidence for mucinlike ligand activity distinct from P-selectin glycoprotein ligand-1

Carroll L. Ramos, McRae J. Smith, Karen R. Snapp, Geoffrey S. Kansas, George W. Stickney, Klaus Ley, Michael B. Lawrence*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

32 Scopus citations

Abstract

Recent reports have shown that leukocyte-leukocyte adhesion is dependent on L-selectin end that leukocyte recognition of L-selectin may be mediated by P-selectin glycoprotein ligand-1 (PSGL-1). We show that the specific attachment and rolling of human neutrophils and the leukemia cell lines HL- 60 and U937 on immobilized, purified L-selectin under continuous shear stress is only partially inhibited by treatment with the PSGL-1 monoclonal antibody (MoAb), KPL1 (41% to 53% inhibition), suggesting that L-selectin ligand activity in addition to PSGL-1 may mediate myeloid cell rolling on L- selectin. K562 cells cotransfected with cDNAs encoding α(1,3)fucosyltransferase-VII (FucT-VII) and PSGL-1 rolled on L-selectin. Adhesion of FucT-VII-PSGL-1 transfectants to L-selectin was completely blocked by MoAb KPL1, indicating that both L-selectin and P-selectin bind similar sites on PSGL-1. In support of existence of a non-PSGL-1 L-selectin ligand activity on leukocytes, an HL-60 membrane preparation immunodepleted of PSGL-1 supported rolling of L-selectin, but not P-selectin transfectants. Treatment of HL-60 cells with O-sialoglycoprotein endopeptidase inhibited attachment and rolling on L-selectin and P-selectin. However, neuraminidase treatment completely/blocked HL-60 rolling on L-selectin, but not P- selectin, suggesting L-selectin and P-selectin ligand activities have different contributions of sialic acid. These findings indicate that myeloid cells express sialylated, O-linked glycoprotein ligand activity independent of PSGL-1 that supports L-selectin-mediated rolling.

Original languageEnglish (US)
Pages (from-to)1067-1075
Number of pages9
JournalBlood
Volume91
Issue number3
DOIs
StatePublished - Feb 1 1998

ASJC Scopus subject areas

  • Biochemistry
  • Immunology
  • Hematology
  • Cell Biology

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