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Blood, Vol. 94 No. 8 (October 15), 1999:
pp. 2725-2734
Circulating Activated Platelets Assist THP-1 Monocytoid/Endothelial
Cell Interaction Under Shear Stress
Gregor Theilmeier,
Tim Lenaerts,
Claude Remacle,
Désiré Collen,
Jos Vermylen, and
Marc F. Hoylaerts
From the Center for Molecular and Vascular Biology, Katholieke
Universiteit Leuven, Leuven, Belgium; and the Laboratoire de Biologie
Cellulaire, Universite Catholique de Louvain, Louvain-la-Neuve,
Belgium.
Circulating complexes of leukocytes and activated platelets are
markers for atherosclerosis, but their interaction with the arterial
endothelial lining has not been studied. Therefore, the effect of
activated platelets on rolling and adhesion of labeled human THP-1
monocytoid cells to human umbilical vein endothelial cell (HUVEC)
monolayers was studied by epifluorescence microscopy in a parallel
plate flow chamber. In the absence of activated platelets, THP-1
rolling on resting HUVEC was negligible at shear rates greater than 300 s 1. Activation of HUVEC with 100 nmol/L phorbol
myristate acetate (PMA) increased THP-1 cell adhesion at
shear rates less than 400 s1. Therefore, a shear rate of
400 s1 was identified as a threshold for THP-1 adhesion.
THP-1 rolling on activated HUVEC was reduced by 64% after L-selectin
inhibition but was not affected by P-selectin inhibition. The addition
of 1 to 50 thrombin receptor-activating peptide
(TRAP)-activated platelets per THP-1 cell enhanced
interactions between THP-1 cells and HUVEC, resulting in a steep
bell-shaped dose-response curve, with a peak of 10 ± 3 rolling
cells/50 seconds at 3 platelets per THP-1 cell (P < .01 v control) with a concomitant 2- to 3-fold increase of firmly
adhering cells (P < .01 v control). In reconstituted blood, low numbers of activated platelets had the same effect on THP-1
rolling and adhesion. P-selectin inhibition reduced platelet/THP-1 cell
interaction in suspension and deposition of the complexes on the
endothelial monolayer. Inhibition of both P- and L-selectin reduced
THP-1/HUVEC interactions to 14% (P < .01, n = 4).
Sialidase digestion and removal of terminal sialic acid residues from
HUVEC or THP-1 cells but not from platelets abolished the platelet
mediated augmentation of THP-1 cell adhesion. Thus, THP-1 rolling on
HUVEC is shear-dependent and largely mediated by L-selectin. P-selectin expressed on activated platelets increases monocytoid cell adhesion to
endothelial cells at shear rates found in coronary arteries through
interactions with both endothelial and monocytoid cells and may
facilitate macrophage accumulation in the vessel wall.
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