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* Institute of Cell Biology, ZMBE, University of Münster, D-48149 Münster, Germany; and L-Selectin on neutrophils as well as inducible
E- and P-selectin on endothelium are involved in the
recruitment of neutrophils into inflamed tissue. Based
on cell attachment assays, L-selectin was suggested to
function as a carbohydrate presenting ligand for E- and
P-selectin. However, previous affinity isolation experiments with an E-selectin-Ig fusion protein had failed to
detect L-selectin among the isolated E-selectin ligands
from mouse neutrophils. We show here that L-selectin
from human neutrophils, in contrast to mouse neutrophils, can be affinity-isolated as a major ligand from total cell extracts using E-selectin-Ig as affinity probe. Binding of human L-selectin to E-selectin was direct,
since purified L-selectin could be reprecipitated with
E-selectin-Ig. Recognition of L-selectin was abolished
by sialidase-treatment, required Ca2+, and was resistant
to treatment with endoglycosidase F. Binding of L-selectin to a P-selectin-Ig fusion protein was not observed. In agreement with the biochemical data, the anti-Lselectin mAb DREG56 inhibited rolling of human
neutrophils on immobilized E-selectin-Ig but not on
P-selectin-Ig. No such inhibitory effect was seen with the anti-mouse L-selectin mAb MEL14 on mouse neutrophils. Rolling of E-selectin transfectants on purified
and immobilized human L-selectin was inhibited by
mAb DREG56. We conclude that L-selectin on human
neutrophils is a major glycoprotein ligand among very few glycoproteins that can be isolated by an E-selectin
affinity matrix. The clear difference between human
and mouse L-selectin suggests that E-selectin-binding
carbohydrate moieties are attached to different protein
scaffolds in different species.
Sandoz Pharma Ltd., CH-4002
Basel, Switzerland
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