Inhibition of interleukin-1 beta production by SKF86002: evidence of two sites of in vitro activity and of a time and system dependence.
Cytokine-suppressing anti-inflammatory drugs (CSAIDs) are reported to inhibit production of proinflammatory cytokines such as interleukin-1 beta (IL-1 beta) and tumor necrosis factor-alpha (TNF-alpha) by affecting a stress-induced kinase. To gain a better understanding of the selectivity and cellular dynamics of this type of inhibitor, we studied in vitro the prototype member of this class of agents, SKF86002. Lipopolysaccharide (LPS)-activated human monocytes treated with SKF86002 produced less proIL-1 beta but normal amounts of the noncytokine lysozyme. Two-dimensional gel analysis indicated that only eight polypeptides produced by monocytes were decreased by SKF86002. Inhibition of IL-1 beta production was achieved by affecting two separate steps in this cytokine's biogenesis. First, SKF86002 lowered proIL-1 beta synthesis. By pulse-chase analysis, this effect was localized to a posttranscriptional site of action; maximal inhibition was observed when SKF86002 was added at the time of cytokine translation. Exposure of monocytes to SKF86002 for > 2 hr led to a loss of IL-1 beta inhibitory activity, suggesting that these cells adapted to this agent. Moreover, LPS-activated monocytes that were pretreated with granulocyte-macrophage colony-stimulating factor were less sensitive to the proIL-1 beta inhibitory effect of SKF86002, and production of proIL-1 beta by cytokine-stimulated human fibroblasts was impaired only modestly by the CSAID. A second effect of SKF86002 was to inhibit release of IL-1 beta into the medium in response to high concentrations of LPS; this effect is observed only with freshly isolated human monocytes as other IL-1 beta-producing cells do not release significant cytokine in response to LPS. The ability of SKF86002 to inhibit this posttranslational mechanism was mimicked by lysosomotrophic agents such as chloroquine, quinacrine, and methylamine. In contrast, chloroquine, and quinacrine were not effective inhibitors of monocyte proIL-1 beta translation. Thus, SKF86002 inhibits IL-1 beta production by affecting at least two distinct steps in the biosynthesis of this cytokine. Manifestation of these two effects, however, is dependent on the length of time for which cells are exposed to this agent and the nature of the cytokine-producing cellular system.
Perregaux
D G
DG
Department of Cancer, Pfizer Inc., Groton, Connecticut 06340, USA.
Dean
D
D
Cronan
M
M
Connelly
P
P
Gabel
C A
CA
D000894:Anti-Inflammatory Agents, Non-Steroidal / Q000494:pharmacology*; D002478:Cells, Cultured; D005347:Fibroblasts / Q000187:drug effects / Q000378:metabolism; D006801:Humans; D007093:Imidazoles / Q000494:pharmacology*; D007375:Interleukin-1 / Q000037:antagonists & inhibitors / Q000096:biosynthesis* / Q000378:metabolism; D007700:Kinetics; D007963:Leukocytes, Mononuclear / Q000187:drug effects / Q000378:metabolism; D008070:Lipopolysaccharides / Q000494:pharmacology; D014176:Protein Biosynthesis; D013844:Thiazoles / Q000494:pharmacology*
D000894:Anti-Inflammatory Agents, Non-Steroidal; D007093:Imidazoles; D007375:Interleukin-1; D008070:Lipopolysaccharides; D013844:Thiazoles; C052026:6-(4-fluorophenyl)-2,3-dihydro-5-(4-pyridinyl)imidazo(2,1-b)thiazole