Stimulation of macrophages with IgG2 immune complexes induced dose-dependently the O
2- generation and the release of arachidonic acid and its metabolites. This Fc
γR-mediated O
2- generation was inhibited by a phospholipase A
2 inhibitor, 4-
p-bromophenacyl bromide (4-pBPB), in parallel to the dose-dependent inhibition of arachidonic acid release. The main arachidonic acid metabolites released were shown to be prostaglandin E
2 and thromboxane B
2 and blocking of the production of these metabolites by indomethacin did not inhibit the O
2- generation. Inhibition of the Fc
γR-mediated O
2- generation and the arachidonic acid release by the C-kinase inhibitor, 1-(5-isoquinolinesulfonyl)-2-methylpiperazine (H-7), was less intense than by 4-pBPB. These results support the previously proposed hypothesis that arachidonic acid acts as an intracellular activator of the Fc
γR-mediated O
2- generation in macrophages. Although the C-kinase activation may also contribute to the activation of the O
2--generating system, arachidonic acid release appears to play a major role in Fc
γR-mediated O
2- generation. In contrast, activation of C-kinase seems to be contributing mainly in the induction of both the arachidonic acid release and O
2- generation by 12-
o-tetradecanoylphorbol 13-acetate (TPA). Furthermore, suboptimal concentrations of TPA and arachidonate were found to act synergistically to stimulate O
2- generation and the inhibition study suggested a positive synergism between C-kinase and arachidonic acid release to induce O
2- generation. This synergistic action may have general importance in receptor-mediated O
2- generation.
View full abstract