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Effect of venous shear stress on CD18-mediated neutrophil adhesion to
cultured endothelium
MB Lawrence, CW Smith, SG Eskin and LV McIntire
Biomedical Engineering Laboratory, Rice University, Houston, TX 77251.
The CD11/CD18 family of glycoproteins has been identified as a mediator of
a number of adhesive interactions crucial to inflammatory responses. Using
a monoclonal antibody (MoAb) against CD18 (TS1/18), the role of these
molecules in polymorphonuclear neutrophil (PMNL) adhesion to cultured
primary human umbilical vein endothelial cells (HUVEC) was examined under
venous flow conditions. Incubation of PMNL with TS1/18 (anti-CD18) did not
inhibit PMNL adhesion to interleukin-1 (IL-1)- treated HUVEC at 2.0
dynes/cm2 (TS1/18-treated 305 +/- 58 PMNL/mm2 v 334 +/- 63 PMNL/mm2 on
control). Furthermore, incubation of HUVEC with R6.5.D6, an MoAb against
intercellular adhesion molecule-1 (ICAM-1) did not significantly inhibit
PMNL adhesion to IL-1-treated HUVEC at 2.0 dynes/cm2 (P greater than .3).
In contrast to the lack of inhibition of adhesion under conditions of flow,
incubation of PMNL with TS1/18 reduced PMNL adherence in static adhesion
assays. PMNL migration beneath HUVEC monolayers has been shown to be
stimulated by 4-hour IL-1 treatment. TS1/18 and R6.5.D6 significantly
inhibited migration of PMNL beneath IL-1-treated HUVEC monolayers under
flow conditions by slightly more than 80% (P less than .005). In flow
experiments with CD18- deficient PMNL, virtually no transendothelial
migration was observed. The effect of FMLP (10(-8) mol/L) on PMNL adhesion
to untreated HUVEC at wall shear stresses ranging from 0.25 to 2.0
dynes/cm2 was also investigated. FMLP had little effect on PMNL adherence
at shear stresses above 0.5 dynes/cm2 (P greater than .45). In response to
FMLP exposure at lower wall shear stresses, PMNL adherence to untreated
HUVEC increased 6.9-fold at 0.5 dynes/cm2 (P less than .001). At 0.25
dynes/cm2, FMLP stimulation increased PMNL adherence to untreated HUVEC
6.5-fold compared with controls (P less than .005), and FMLP failed to make
CD18-deficient PMNL more adherent. In experiments with PMNL pretreated with
TS1/18 (anti-CD18), there was a 67% inhibition of FMLP- stimulated adhesion
at 0.5 dynes/cm2 (P less than .025). The upper threshold of CD18-mediated
PMNL adhesion appears to be between 0.5 and 1.0 dyne/cm2. Above these wall
shear stresses, the initial attachment of PMNL to cultured endothelium was
mediated almost exclusively by CD18- independent mechanisms. By simulating
some of the flow parameters in the microcirculation with well-characterized
shear forces, PMNL adhesion by CD18-independent and dependent mechanisms
can be differentiated. These data also indicate that CD18 is an important
mediator of transendothelial migration by PMNL, which have attached to the
endothelium by a CD18-independent mechanism.
Volume 75,
Issue 1,
pp. 227-237,
01/01/1990
Copyright © 1990 by The American Society of Hematology
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