PROTEIN-KINASE-C-INDEPENDENT ACTIVATION OF ARACHIDONATE RELEASE AND PROSTAGLANDIN-E2 FORMATION IN MACROPHAGES INTERACTING WITH CERTAIN BACTERIA

Certain bacterial species, of which we selected Fusobacterium nucleatum, Gardnerella vaginalis, Peptostreptococcus anaerobius and Propionibacterium acnes, were found to induce release of arachidonic acid in a dose- and time-dependent manner in mouse macrophages. The release of arachidonic acid showed a characteristic lag period of approximately 10 min and was accompanied by selective transformation into prostaglandin E2. Bacteria killed by various methods caused a similar response, indicating that bacterial surface structures rather than secreted products were involved. Down-regulation of protein kinase C by treatment of macrophages with 4-beta-phorbol 12-myristate 13-acetate hardly affected the response to bacteria at all, except for a partial inhibition in the case of P. acnes. Furthermore, the generation of prostaglandin E2 was synergistically enhanced when macrophages were exposed to both bacteria and phorbol ester. It is also unlikely that bacterial activation was mediated exclusively via a rise in cytosolic [Ca2+], since bacteria stimulated the release of arachidonic acid also when [Ca2+] was clamped at various levels and since the response to bacteria was enhanced in an additive to synergistic manner when combined with calcium ionophore. Changes in protein phosphorylation in macrophages exposed to F. nucleatum (Gram-negative) were virtually identical to those seen with bacterial lipopolysaccharide, while P. anaerobius (Gram-positive) induced enhanced labeling of a single detectable phosphoprotein. In both cases, the changes in protein phosphorylation showed a time lag of 4-8 min and occurred independently of protein kinase C, consistent with a possible role in signal transduction. These results demonstrate that certain bacteria cause activation of arachidonic acid release and prostaglandin E2 formation in mouse macrophages; that the response is independent of protein kinase C and that it is not wholly mediated via a rise in cytosolic [Ca2+].