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P-Selectin Support of Neonatal Neutrophil Adherence Under Flow:
Contribution of L-Selectin, LFA-1, and Ligand(s) for P-Selectin
M. Michele Mariscalco,
M. Hossein Tcharmtchi, and
C. Wayne Smith
From the Sections of Leukocyte Biology and Critical Care Medicine,
Department of Pediatrics, Baylor College of Medicine, Houston, TX.
To further define the neonatal neutrophil's ability to localize to
inflamed tissue compared with adult cells, we examined the neonatal
neutrophil interactions with P-selectin monolayers under two
conditions: (1) attachment under constant shear stress and flow and (2)
detachment where cells were allowed to attach in the absence of shear
stress and then shear stress is introduced and increased in step-wise
increments. Cord blood and adult neutrophils had minimal interactions
with unstimulated human umbilical vein endothelial cells (HUVECs) at a
constant shear stress of 2 dynes/cm2. There was a marked
increase in the number of both neonatal and adult cells interacting
(interacting cells = rolling + arresting) with HUVECs after
histamine stimulation, although the neonatal value was only 40% of
adult (P < .05). Neonatal neutrophils also had significantly
decreased interaction with monolayers of Chinese hamster ovary (CHO)
cells transfected with human P-selectin (CHO-P-selectin; 60% of adult
values, P < .003). Of the interacting cells, there was a
lower fraction of neonatal cells that rolled compared with adult cells
on both stimulated HUVECs and CHO-P-selectin. That neonatal neutrophil
L-selectin contributes to the diminished attachment to P-selectin is
supported by the following: (1) Neonatal neutrophils had significantly
diminished expression of L-selectin. (2) Anti-L-selectin monoclonal
antibody reduced the number of interacting adult neutrophils to the
level seen with untreated neonatal neutrophils, but had no effect on
neonatal neutrophils. In contrast, L-selectin appeared to play no role
in maintaining the interaction of either neonatal or adult neutrophils
in the detachment assay. Once attachment occurred, the neonatal
neutrophil's interaction with the P-selectin monolayer was dependent
on LFA-1 and to other ligands to a lesser degree based on the
following: (1) Control neonatal neutrophils had decreased rolling
fraction compared with adult neutrophils, although the total number of
interacting neutrophils was equal between groups. (2) Anti-LFA-1
treatment resulted in an increase in the rolling fraction of both
neonatal and adult neutrophils. However, whereas the number of
interacting adult neutrophils remained unchanged, the number of
neonatal neutrophils decreased with increased shear stress. We
speculate that this increased detachment of neonatal cells is due to
differences in neutrophil ligand(s) for P-selectin.
Blood, Vol. 91 No. 12 (June 15), 1998:
pp. 4776-4785
© 1998 by The American Society of Hematology.
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