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-Activated Endothelium Under Flow In Vitro1
*
Vascular Research Division, Departments of Pathology, Brigham and Women’s Hospital and Harvard Medical School, Boston, MA 02115;
Department of Immunology and Microbiology, Northwestern University School of Medicine, Chicago, IL 60611; and
Drug Discovery Group, Genetics Institute, Cambridge, MA 02140
In this study, an in vitro flow model and a blocking mAb to
P-selectin glycoprotein ligand-1 (PSGL-1) were used to define the role
of PSGL-1 in monocyte attachment and rolling on E- and P-selectin and
in attachment and accumulation on 6-h TNF-
-activated HUVEC. KPL1, an
adhesion-blocking mAb directed against the tyrosine sulfate motif of
PSGL-1, abolished monocyte-adhesive interactions with P-selectin, but
only partially blocked monocyte interaction with E-selectin. Further
analysis showed that on E-selectin, KPL1 blocked only secondary (i.e.,
monocyte/monocyte) interactions, but did not block primary (i.e.,
monocyte/E-selectin) interactions, with secondary adhesion accounting
for 90% of the total adhesive interactions on either E- or P-selectin.
On cytokine-activated HUVEC, monocytes initially attached and formed
linear strings of adherent cells, which involved both primary and
secondary adhesion. PSGL-1 or L-selectin mAb reduced string formation,
and the combination of PSGL-1 and L-selectin mAb prevented monocyte
strings and inhibited 86% of accumulation. Monocyte attachment and
rolling on purified adherent monocytes were also critically dependent
on PSGL-1 on the adherent monocytes. These studies document that
secondary interactions between monocytes, mediated by PSGL-1, are
crucial for monocyte initial attachment, rolling, and accumulation on
activated endothelium under laminar shear flow.
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