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Macrophage colony-stimulating factor-induced bone marrow macrophages do not
synthesize or release prostaglandin E2
Y Shibata, DR Bjorkman, M Schmidt, Y Oghiso and A Volkman
Department of Pathology and Laboratory Medicine, East Carolina University
School of Medicine, Greenville, NC.
Previously, we found that murine bone marrow-derived macrophages (MO)
induced in vitro by MO-specific colony-stimulating factor (M-CSF) have
little capacity to release prostaglandin E2 (PGE2) and other eicosanoids.
This work focused on the functional and transcriptional expression of the
key enzymes for the PGE2 synthesis in the MO. Nonadherent bone marrow cells
were cultured with RPMI1640 plus 10% fetal bovine serum (FBS) further
supplemented with either M-CSF or granulocyte-macrophage (GM)-CSF and
interleukin-3 (IL-3). Cellular PGG/H synthase (cyclooxygenase) levels were
quantified by cytometric analysis with antibodies specific for the two
isozymes of PGG/H synthase (PGG/H synthases 1 and 2). The enzyme activity
was monitored by adding exogenous arachidonic acid (AA) substrate to the
bone marrow MO cultures and to the cell-free particulate fractions. The
levels of PGE2 converted were quantitated by radioimmunoassay (RIA). mRNA
levels of the enzymes were detected by Northern blot analysis hybridized
with mouse PGG/H synthase cDNA probes, 2.7 kb (PGG/H synthase 1) and 4.2 kb
(PGG/H synthase 2). In addition, cellular phospholipase A2 (PLA2)
activities were detected with sn-2-14C-arachidonyl phosphatidylcholine as a
substrate. Cells proliferating in the presence of GM-CSF and IL-3 for more
than 4 days showed significant release of PGE2 (> 7 ng/10(6) cells) when
stimulated by AA. These cells also expressed significant amounts of PGG/H
synthase 1 protein, its mRNA (2.7 kb) and cellular PLA2. M-CSF-induced MO,
in sharp contrast, expressed little PGG/H synthase protein, mRNA, cellular
enzyme activity, or PGE2 release, despite comparable levels of cellular
PLA2 activity. These data suggest that the capacity of differentiating
marrow-derived MO to form PGE2 is growth factor-dependent.
Volume 83,
Issue 11,
pp. 3316-3323,
06/01/1994
Copyright © 1994 by The American Society of Hematology
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