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(Received for publication, April 18, 1996, and in revised form, October 5, 1996)
From the Department of Immunopharmacology, SmithKline Beecham
Pharmaceuticals, King of Prussia, Pennsylvania 19406
Human monocytes possess several acylhydrolase
activities and are capable of producing both prostanoids (PG) and
leukotriene (LT) products upon acute stimulation with calcium
ionophore, A23187 or phagocytosis of zymosan particles. The cytosolic
85-kDa phospholipase (PLA) A2 co-exists with the 14-kDa
PLA2 in the human monocyte, but their respective roles in
LT production are not well understood. Reduction in 85-kDa
PLA2 cellular protein levels by initiation site-directed
antisense (SK 7111) or exposure to the 85-kDa PLA2 inhibitor, arachidonyl trifluoromethyl ketone (AACOCF3),
prevented A23187 or zymosan-stimulated monocyte prostanoid formation.
In contrast, neither treatment altered stimulated LTC4
production. This confirmed the important role of the 85-kDa
PLA2 in prostanoid formation but suggests that it has less
of a role in LT biosynthesis. Alternatively, treatment of monocytes
with the selective, active site-directed 14-kDa PLA2
inhibitor, SB 203347, prior to stimulation had no effect on prostanoid
formation at concentrations that totally inhibited LT formation.
Addition of 20 µM exogenous arachidonic acid to monocytes
exposed to SK 7111 or SB 203347 did not alter A23187-induced
PGE2 or LTC4 generation, respectively,
indicating that these agents had no effect on downstream arachidonic
acid-metabolizing enzymes in this setting. Taken together, these
results provide evidence that the 85-kDa PLA2 may play a
more significant role in the formation of PG than LT. Further,
utilization of SB 203347 provides intriguing data to form the
hypothesis that a non-85-kDa PLA2 sn-2 acyl
hydrolase, possibly the 14-kDa PLA2, may provide substrate
for LT formation.
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