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Blood, Vol. 92 No. 12 (December 15), 1998:
pp. 4691-4699
Differential Leukocyte Recruitment From Whole Blood Via Endothelial
Adhesion Molecules Under Shear Conditions
Paul H. Reinhardt and
Paul Kubes
From the Immunology Research Group, University of
Calgary, Calgary, Alberta, Canada.
The objective of this study was to determine if vascular cell
adhesion molecule (VCAM-1), E-selectin, and P-selectin could selectively recruit leukocyte subpopulations, and whether this was
affected by shear force or adhesion molecule concentration. Cover slips
coated with purified adhesion molecules were incorporated into laminar
flow chambers. Whole human blood was perfused for 5 minutes over these
cover slips at relative shear forces of 2 to 40 dynes/cm2.
Chasing the whole blood with buffer permitted visualization of
leukocyte-substratum interactions. Leukocytes were observed to roll on
and adhere to VCAM-1 at shears between 2 and 15 dynes/cm2.
As assessed by cover slip staining, the majority of these cells were
lymphocytes, but eosinophils, monocytes, and, surprisingly, neutrophils
were also recruited, events inhibitable by
anti- 4-integrin antibody (HP1/2). Neutrophils were
effectively recruited onto the selectins, with interactions occurring
at shears as high as 30 and 40 dynes/cm2 for E- and
P-selectin respectively. Eosinophils had high affinity for P- but not
E-selectin. Mononuclear cells did not have high affinity for either
selectin, but interacted avidly with VCAM-1. Antibodies against
P-selectin (G1) and E-selectin (ES-1) completely blocked interactions
on these substrates. Reducing the concentration of adhesion molecules
did not appreciably change recruitment patterns except for VCAM-1,
where neutrophils were no longer recruited. The novel use of whole
blood in flow chambers shows a partial selectivity of selectins and
VCAM-1 for certain subpopulations of leukocytes under varying
physiologic shear conditions.
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