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Departments of
*
Pathology and
Surgery, University of Florida, Gainesville, FL 32610
PG added to cell culture profoundly affect the in vitro maturation
and function of monocyte-derived dendritic cells (MDC). Because
unstimulated monocytes express cyclooxygenase (COX)-1, and COX-2 when
activated, we examined whether MDC express these enzymes and produce
prostanoids that autoregulate maturation and IL-12 production. Immature
MDC (I-MDC) and mature MDC express COX-1, but, unlike monocytes, both
MDC populations constitutively express COX-2. However, COX-2 regulation
in both MDC populations differs from monocytes, as IL-4 does not
suppress enzyme expression. COX-2 is functional in MDC as a specific
inhibitor, NS-398, significantly reduces PGE2 production.
I-MDC undergoing maturation with soluble CD40 ligand (sCD40L) increase
PGE2 synthesis, but prostanoid synthesis is switched to
COX-1. However, with IFN-
present, sCD40L-stimulated PG metabolism
is redirected to COX-2, and PGE2 synthesis increases
severalfold. Endogenous PG production by MDC does not regulate CD40,
CD80, CD86, or HLA DR expression; however, it does promote MDC
maturation, as NS-398 significantly reduces CD83 expression in I-MDC
matured with sCD40L/IFN-. PG produced through COX-2 also
autoregulate IL-12, but the effects are dependent on the MDC maturation
state. Blocking COX-2 reduces I-MDC secretion of IL-12p40, whereas it
increases IL-12p40 and p70 production by maturing MDC. COX-2-mediated
PG production impacts MDC function as maturing these cells in the
presence of NS-398 yields MDC that stimulate significantly more IFN-
in an allogeneic mixed lymphocyte response than MDC matured without
this inhibitor. These studies demonstrate that MDC express both COX
isoforms constitutively and produce prostanoids, which autoregulate
their maturation and function.
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