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Vol. 280, Issue 2, 1065-1074, 1997
in Human Amnionic WISH Cells by Various Stimuli Occurs
Through Distinct Intracellular Mechanisms
Immunosciences Research Area (K.I.H., E.R.O., J.L., R.L.B., G.W.C.,
K.B.G.) and the
Aging and Degenerative Diseases Research Area
(B.W.E., D.J.C.), Abbott Laboratories, Abbott Park, Illinois
These studies examined the signal transduction mechanisms by which
prostaglandin (PG) E2 production can occur in human
amnionic WISH cells in response to the stimuli okadaic acid,
interleukin (IL)-1
, tumor necrosis factor (TNF)-
,
phorbol-12-myristate-13-acetate (PMA) or combinations of PMA with
IL-1 or TNF-. We also investigated whether WISH cells are capable
of producing TNF- or IL-1 in response to stimulation, because
these cytokines can be produced in an autocrine fashion to perpetuate
an inflammatory response. Our data indicate that the magnitude of
PGE2 production induced by a given stimulus correlated
temporally with the level of PGH synthase-2 (PGHS-2) protein. PMA or
IL-1 induced PGE2 production 2 to 4 hr after treatment,
whereas the combination of these agents produced the most rapid
induction 2 hr after treatment. Only okadaic acid induced the
production of both PGE2 and TNF-, after a lag of 12 to
18 hr. PGE2 production by all stimuli was inhibited by dexamethasone, the IL-1 receptor antagonist (IL-1ra), the specific PGHS-2 inhibitor NS-398 and the protein kinase inhibitor staurosporin. In contrast, TNF- production in response to okadaic acid was inhibited by the TNF-converting enzyme inhibitor GI 129471 and staurosporin but was unaffected by either IL-1ra, dexamethasone or
NS-398. We conclude that WISH cells are capable of producing bioactive
proinflammatory mediators such as TNF- and PGE2 through separable intracellular signal transduction mechanisms. The ability of
IL-1ra to reduce PGE2 production caused by all stimuli used suggests an autocrine role for IL-1 in PGHS-2 induction in these cells.
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