TRANSCRIPTION, TRANSLATION AND SECRETION OF INTERLEUKIN-1 AND TUMOR-NECROSIS-FACTOR - EFFECTS OF TEBUFELONE, A DUAL CYCLOOXYGENASE/5-LIPOXYGENASE INHIBITOR

We examined the effect of tebufelone, a dual cyclooxygenase (CO)/5-lipoxygenase (LO) inhibitor, on the synthesis, secretion and gene expression of interleukin (IL) 1-beta and tumor necrosis factor (TNF)-alpha by human peripheral blood mononuclear cells (PBMC). Basal concentrations of immunoreactive IL 1-beta and TNF-alpha after 18-24 h, in the absence or presence of tebufelone (less-than-or-equal-to 12.5-mu-M), were near the limit of detection (100 pg/ml). By contrast, preincubation (1 h) of cells, in amounts of tebufelone which decrease the formation of leukotriene (LT) B4, markedly enhanced (up to 500%) the synthesis of IL 1-beta and TNF-alpha following lipopolysaccharide (LPS), heat-killed Staphylococcus epidermidis or concanavalin A stimulation. Moreover, a disproportionate amount of the overall increase in IL 1 (alpha and beta) was secreted in contrast to the amount which remained cell associated, an effect unrelated to cell damage or leakage as tubefelone had no effect on either lactate dehydrogenase release by PBMC, or mitochondrial dehydrogenases of adherent monocytes as detected by enzymatic cleavage of the substrate 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide. There was no inverse correlation between the changes in prostaglandin (PG)E2 levels and TNF-alpha or IL 1-beta synthesis, and when PG formation was maximally inhibited by preincubating the cells in indomethacin, tubufelone, added 1 h before the stimulus, continued to enhance the synthesis of IL 1-beta although not that of TNF-alpha. The addition of the CO/5-LO inhibitor 2 h after LPS stimulation, however, did not interfere with IL 1-beta synthesis, suggesting that tebufelone interacts with an early event(s)in the activation of PBMC. For IL 1-beta and TNF-alpha, basal and stimulated (4 h post LPS) mRNA levels were not increased by by tebufelone, despite a concomitant increase in the synthesis of IL 1-beta. In conclusion, we have demonstrated that tebufelone enhances IL 1(alpha and beta) and TNF-alpha synthesis at concentrations which suppress leukotriene formation. These findings argue against a role of 5-LO products as necessary intermediates of IL 1 (alpha and beta) and TNF-alpha synthesis.