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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, and
Michael B. Lawrence
From the Department of Biomedical Engineering, University of Virginia
Health Sciences Center, Charlottesville; and the Department of
Microbiology-Immunology, Northwestern University Medical School,
Chicago, IL.
Recent reports have shown that leukocyte-leukocyte adhesion is
dependent on L-selectin and 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  (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.
Blood, Vol. 91 No. 3 (February 1), 1998:
pp. 1067-1075
© 1998 by The American Society of Hematology.
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