Macrophage Colony - Stimulating Factor - Induced Bone Marrow Macrophages Do Not Synthesize

Previously, we found that murine bone marrow-derived macrophages (M01 induced in vitro by M0-specific colonystimulating factor (M-CSF) have little capacity to release prostaglandin E, (PGE,) and other eicosanoids. This work focused on the functional and transcriptional expression of the key enzymes for the PGE, synthesis in the MO. Nonadherent bone marrow cells were cultured with RPM1 1640 plus 10Y0 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 IPGG/H synthases 1 and 2). The enzyme activity was monitored by adding exogenous arachidonic acid (AA) substrate to the bone marrow M0 cultures and to the cell-free particulate fractions. The levels of PGE, converted were quantitated by radioimmunoassay (RIA). mRNA leveis of the enzymes