Effect of protein kinase C inhibitor (H-7) and calmodulin antagonist (W-7) on pertussis toxin-induced IL-1 production by human adherent monocytes. Comparison with lipopolysaccharide as a stimulator of IL-1 production.
Human adherent monocytes stimulated with 1 microgram/ml pertussis toxin (PT) produced interleukin-1 (IL-1), as measured by thymocyte co-stimulation assay and enzyme-linked immunosorbent assay (ELISA), specific for IL-1 alpha and IL-1 beta. To clarify the role of protein kinase C (PKC) and calmodulin in IL-1 production, we investigated the effects of a PKC inhibitor, H-7, and a calmodulin antagonist, W-7 on PT- and lipopolysaccharide (LPS)-induced IL-1 production by monocytes. Addition of 10 microM and 20 microM H-7 to the culture medium markedly suppressed both PT- and LPS-induced IL-1 production. PT-induced IL-1 production was significantly suppressed by 5 microM and 10 microM W-7. However, LPS-induced IL-1 production was not suppressed by W-7 at the concentrations tested. When monocytes were labelled with Quin 2/AM, IL-1 production by monocytes stimulated with PT and LPS was markedly suppressed. These results indicate that different pathways are involved in the IL-1 production by PT and LPS; both calmodulin- and PKC-dependent processes are necessary for the IL-1 production induced by PT, whereas LPS-induced IL-1 production is dependent on the PKC. Inhibition of IL-1 production by interfering with intracellular Ca2+ trafficking in Quin 2/AM-loaded monocytes may be associated with the inhibition of PKC and calmodulin activity.
Taniguchi
H
H
Department of Pediatrics, Hiroshima University School of Medicine, Japan.
Sakano
T
T
Hamasaki
T
T
Kashiwa
H
H
Ueda
K
K
D019307:1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine; D000634:Aminoquinolines; D000818:Animals; D002147:Calmodulin / Q000037:antagonists & inhibitors / Q000502:physiology*; D002478:Cells, Cultured; D002614:Chelating Agents; D005456:Fluorescent Dyes; D007375:Interleukin-1 / Q000096:biosynthesis*; D007546:Isoquinolines / Q000494:pharmacology*; D008070:Lipopolysaccharides; D008297:Male; D051379:Mice; D008809:Mice, Inbred C3H; D009000:Monocytes / Q000187:drug effects* / Q000378:metabolism*; D037342:Pertussis Toxin; D010879:Piperazines / Q000494:pharmacology*; D011493:Protein Kinase C / Q000037:antagonists & inhibitors / Q000502:physiology*; D013449:Sulfonamides / Q000494:pharmacology*; D010566:Virulence Factors, Bordetella
D000634:Aminoquinolines; D002147:Calmodulin; D002614:Chelating Agents; D005456:Fluorescent Dyes; D007375:Interleukin-1; D007546:Isoquinolines; D008070:Lipopolysaccharides; D010879:Piperazines; D013449:Sulfonamides; D010566:Virulence Factors, Bordetella; C017967:W 7; C035738:Quin2-acetoxymethyl ester; D019307:1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine; D037342:Pertussis Toxin; D011493:Protein Kinase C