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Department of Physiology, The Medical School, University of Birmingham, Birmingham, United Kingdom
Neutrophils migrate through endothelium using an ordered sequence
of adhesive interactions and activating signals. To investigate the
consequences of disruption of this sequence, we characterized adhesion
and migration of neutrophils perfused over HUVEC that had been treated
with TNF-
for 4 h and evaluated changes caused by exogenously
added chemotactic agents. When HUVEC were treated with 2 U/ml TNF,
flowing neutrophils adhered, with the majority rolling and relatively
few migrating through the monolayer. If fMLP, IL-8, zymosan-activated
plasma (a source of activated complement factor C5a), epithelial
cell-derived neutrophil-activating peptide (ENA-78), or
growth-regulating oncogene, GRO-, was perfused over these
neutrophils, they stopped rolling and rapidly migrated over the
monolayer, but did not penetrate it. When HUVEC were treated with 100
U/ml TNF, the majority of adherent neutrophils transmigrated. If
neutrophils were treated with fMLP, IL-8, C5a, ENA-78, or GRO- just
before perfusion over this HUVEC, transmigration, but not adhesion, was
abolished. However, when platelet-activating factor was used to
activate neutrophils, migration through HUVEC treated with 100 U/ml TNF
was not impaired, and migration through HUVEC treated with 2 U/ml TNF
was actually increased. Transmigration required ligation of CXC
chemokine receptor-2 on neutrophils, and differential desensitization
of this receptor (e.g., by fMLP but not platelet-activating factor) may
explain the pattern of disruption of migration. Thus, transmigration
may require presentation of the correct activators in the correct
sequence, and inappropriate activation (e.g., by systemic activators)
could cause pathological accumulation of neutrophils in the vessel
lumen.
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