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Blood, Vol. 95 No. 3 (February 1), 2000:
pp. 911-920
Sequential binding of CD11a/CD18 and CD11b/CD18 defines neutrophil
capture and stable adhesion to intercellular adhesion
molecule-1
Eric R. Hentzen,
Sriram Neelamegham,
Geoffrey S. Kansas,
Jennifer A. Benanti,
Larry V. McIntire,
C. Wayne Smith, and
Scott I. Simon
From the Speros Martel Section of Leukocyte Biology, Department of
Pediatrics, Baylor College of Medicine, Houston, TX; The Cox Laboratory
for Biomedical Engineering, Institute of Biosciences and
Bioengineering, Rice University, Houston, TX; Department of Chemical
Engineering, State University of New York, Buffalo, NY; and the
Department of Microbiology and Immunology, Northwestern University
Medical School, Chicago, IL.
The relative contributions of CD11a/CD18 and CD11b/CD18 to the
dynamics and strength of neutrophil adhesion to intercellular adhesion
molecule (ICAM)-1-transfected cells were examined over the time course
of chemotactic stimulation. Suspensions of neutrophils and
transfectants were sheared in a cone-plate viscometer, and formation of
heterotypic aggregates was measured by 2-color flow cytometry. The
2-body collision theory was used to compute adhesion efficiency,
defined as the proportion of collisions between neutrophils and target
cells that resulted in capture. ICAM-1 surface density and shear rate
both regulated adhesion efficiency. Target cells expressing
approximately 1000 ICAM-1 sites/µm2 (Ilow)
were captured with an efficiency of 0.15 at 100 s 1,
which decreased to zero at 300 s1. At 8-fold higher
ICAM-1 expression (Ihigh) corresponding to levels measured
on interleukin-1-stimulated endothelium, efficiency was 0.3 at 100 s1 and remained above background to 900 s1. Shear alone was sufficient for CD11a/CD18-mediated
adhesion to ICAM-1, and stimulation with
formyl-methionyl-leucyl-phenylalanine boosted capture efficiency
through CD11a/CD18 by 4-fold. In comparison, CD11b/CD18 supported one
third of this efficiency, but was necessary for aggregate stability
over several minutes of shear and at shear stresses exceeding 5 dyne/cm2. Hydrodynamics influenced capture efficiency
predominantly through the collisional contact duration, predicted
to be approximately 9 milliseconds for successful capture of
Ilow and 4 milliseconds for Ihigh. The
implication is that an increase in ICAM-1 from resting levels to those
on inflamed endothelium effectively increases the permissible shear in
which capture through  2-integrins may occur. Neutrophil
adhesion to ICAM-1 appears to be a cooperative and sequential process
of CD11a-dependent capture followed by CD11b-mediated stabilization.
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