REGULATION OF ALVEOLAR MACROPHAGE PRODUCTION OF CHEMOATTRACTANTS BY LEUKOTRIENE B4 AND PROSTAGLANDIN-E2

Alveolar macrophages (AM) appear to influence the recruitment of neutrophils into the lung by the elaboration of both lipid and peptide chemotactic molecules for neutrophils. Little is known about the mechanisms that regulate production or release of chemotactic molecules by AM or the interaction between these classes of chemotactic molecules. We investigated the hypothesis that the lipid mediator leukotriene B4 (LTB4) has an in vitro regulatory action on the production of chemotactic proteins by AM. In these experiments, the chemotactic activity in AM culture supernatants was measured in a modified Boyden chamber. LTB4 treatment increased AM production of chemotactic activity in excess of what might be attributed to the amount of LTB4 measured in the culture supernatant after the incubation period. This effect was magnified by in vivo administration of endotoxin prior to AM harvesting. Pretreatment with LTB4 caused a sustained 250% increase in AM production of chemotactic activity, yet only negligible amounts of LTB4 were measured by gas chromatography/mass spectrometry in the LTB4-pretreated AM culture supernatants, indicating that LTB4 alone did not account for the chemotactic activity observed in our studies. A chemotactic peptide in LTB4-treated AM culture supernatant could be isolated and separated from LTB4 by molecular sieve chromatography. Purified column fractions contained 80% of the chemotactic activity of endotoxin-stimulated AM culture supernatant and had a molecular mass of 10,000 D. In contrast to LTB4, prostaglandin E2 (PGE2) suppressed chemotactic activity production by endotoxin-stimulated AM by 70%. Pretreatment with PGE, was not effective; PGE2 had to be present in the AM culture medium during endotoxin exposure in order to exert a suppressive effect. The effect on chemotactic activity production appears to be specific since neither LTB4 or PGE2 altered tumor necrosis factor production by AM. These data indicate that ambient levels of LTB4 and PGE2 may regulate the synthesis, release, or bioactivity of certain chemotactic cytokines by AM.