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B
Immunology Research Group and Departments of Physiology and Biophysics and Medicine, University of Calgary, Calgary, Alberta, Canada T2N 4N1
With the use of a whole blood laminar flow
chamber system, we examined the types of leukocytes, adhesion molecules
and the role of nuclear factor-
B (NF-B) in thrombin-induced
leukocyte recruitment. Primary human umbilical vein endothelial cells
(HUVEC) stimulated with thrombin induced a significant increase in
P-selectin-dependent neutrophil recruitment. Unexpectedly, brief
thrombin stimulation (3 min) of endothelium also induced a significant
lymphocyte recruitment 4 h later in addition to neutrophil
recruitment. E-selectin antibody reduced neutrophil recruitment by
>90%, whereas vascular adhesion molecule-1
(VCAM-1)/
4-integrin were primarily responsible for lymphocyte recruitment. To examine whether NF-B contributed to leukocyte recruitment 4 h post thrombin stimulation, we treated HUVEC with the NF-B inhibitor MG-132 for 1 h before thrombin stimulation. MG-132 significantly reduced the number of rolling (77.1%) and adherent (79.9%) leukocytes compared with thrombin stimulation alone. The inhibitor was more effective at preventing lymphocyte than neutrophil recruitment, consistent with its greater effect on VCAM-1 versus E-selectin expression. Tumor necrosis factor-- and MG-132-treated HUVEC displayed no inhibition of leukocyte recruitment despite a decrease in NF-B activation. In
summary, thrombin causes predominant neutrophil recruitment via rapid
P-selectin expression but also a delayed E-selectin- and
VCAM-1-dependent neutrophil and lymphocyte recruitment via de novo
protein synthesis. Although NF-B mobilization was essential for
thrombin-mediated VCAM-1-dependent recruitment, it only partially contributed to E-selectin-dependent recruitment.
selectins; vascular adhesion molecule-1; integrin; tumor necrosis
factor-; endothelium
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