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Periodontics and Endodontics and
Microbiology and Immunology, Tohoku University Graduate School of Dentistry, Sendai, Japan
Activated polymorphonuclear leukocytes (PMNs) release various types
of proteases and express them on the cell surface. The proteases play
important roles in PMN-mediated events. In the present study, flow
cytometric analysis revealed that CD14 expression on human gingival
fibroblasts (HGF) was markedly reduced by PMA-activated PMNs in a
coculture system. We found that this reduction was caused by both
secreted and cell surface proteases produced by activated PMNs. A
protease responsible for the reduction was found to be human leukocyte
elastase (HLE) secreted from the activated PMNs by use of various
protease inhibitors, although HLE was only partially involved in CD14
reduction caused by cell-bound molecule(s) on fixed PMNs. Analysis with
purified HLE revealed a time- and dose-dependent reduction of CD14 on
HGF, and complete reduction was observed by 20 µg/ml HLE treatment
for 30–60 min, but the other molecules such as CD26, CD59, CD157, and
MHC class I on HGF were only slightly reduced. This reduction of CD14
resulted from direct proteolysis by HLE on the cell surface, because
HLE reduced CD14 on fixed HGF and also on purified cell membranes. As a
result of CD14 proteolysis, IL-8 production by HGF was suppressed when
triggered by 10 ng/ml LPS, but not by IL-1
, indicating that HLE
inhibited a CD14-dependent cell activation. These findings suggested
that activated PMNs have a potential negative feedback mechanism for
HGF function at the inflammatory site, particularly in periodontal
tissues.
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