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Immunopharmacology Group, Southampton General Hospital, Southampton, United Kingdom
Mast cells are found frequently in close proximity to blood
vessels, and endothelial cells are likely to be exposed to high
concentrations of their granule mediators. We have investigated the
proinflammatory actions of the major mast cell product tryptase on
HUVEC. Addition of purified tryptase was found to stimulate thymidine
incorporation, but induced little alteration in cell numbers,
suggesting it is not a growth factor for HUVEC. Expression of ICAM-1,
VCAM-1, and E-selectin was not altered following incubation with
tryptase, but the potent granulocyte chemoattractant IL-8 was released
in a dose-dependent fashion in response to physiologically relevant
concentrations, with maximal levels in supernatants after 24 h.
The actions of tryptase on HUVEC were inhibited by heat inactivation of
the enzyme, or by preincubating with the protease inhibitors leupeptin
or benzamidine, suggesting a requirement for an intact catalytic site.
Reverse-transcription PCR analysis indicated up-regulation of mRNA for
IL-8 as well as for IL-1ß in response to tryptase or TNF-
.
However, tryptase was a more selective stimulus than TNF- and did
not induce increased expression of mRNA for granulocyte-macrophage CSF
or stimulate the release of this cytokine. Leukocyte accumulation in
response to tryptase may be mediated in part through the selective
secretion of IL-8 from endothelial cells.
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