Tryptase activates the mitogen-activated protein kinase/activator protein-1 pathway in human peripheral blood eosinophils, causing cytokine production and release.
We have previously shown that mast cells enhance eosinophil survival and activation. In this study we further characterized mast cell activity toward eosinophils. Sonicate of both rat peritoneal mast cells and the human mast cell line 1 (HMC-1) induced a concentration-dependent IL-6 and IL-8 release from human peripheral blood eosinophils (ELISA). HMC-1-induced IL-8 release was significantly reduced by the tryptase inhibitors GW-45 and GW-58 (90 and 87%, respectively, at an optimal concentration) but not by anti-stem cell factor, anti-TNF-alpha, or anti-IFN-gamma neutralizing Abs or by the antihistamine drugs pyrilamine and cimetidine. In a manner similar to HMC-1, human recombinant tryptase induced the expression of mRNA for IL-8 (RT-PCR) and caused IL-8 release from the eosinophils. Addition of cycloheximide, actinomycin D, dexamethasone, PD 98059, curcumin, or SB 202190 completely inhibited the tryptase-induced IL-6 and IL-8 release. In contrast, cyclosporin A had no effect on tryptase-induced IL-8 release. Tryptase caused phosphorylation of extracellular signal-regulated kinases 1 and 2, c-Jun N-terminal kinases 1 and 2, and p38 (Western blot). Tryptase also induced the translocation of c-Jun from the cytosol to the nucleus (confocal microscopy) and enhanced AP-1 binding activity to the DNA (EMSA). Eosinophils were found to express proteinase-activated receptor 2 (FACS). When eosinophils were incubated with tryptase in the presence of anti-proteinase-activated receptor 2 antagonist Abs a significant decrease in the IL-6 and IL-8 release occurred. In summary, we have demonstrated that the preformed mast cell mediator tryptase induces cytokine production and release in human peripheral blood eosinophils by the mitogen-activated protein kinase/AP-1 pathway.
Department of Pharmacology, School of Pharmacy, Faculty of Medicine, Hebrew University of Jerusalem, Israel.
Vladislav
V
Temkin
Boris
B
Kantor
Vivian
V
Weg
Mor-Li
ML
Hartman
Francesca
F
Levi-Schaffer
D000328:Adult; D000818:Animals; D002460:Cell Line; D002474:Cell-Free System / Q000201:enzymology / Q000502:physiology; D016207:Cytokines / Q000096:biosynthesis* / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D004804:Eosinophils / Q000201:enzymology* / Q000276:immunology / Q000378:metabolism*; D006801:Humans; D015850:Interleukin-6 / Q000096:biosynthesis / Q000378:metabolism; D016209:Interleukin-8 / Q000096:biosynthesis / Q000378:metabolism; D020935:MAP Kinase Signaling System / Q000502:physiology*; D008407:Mast Cells / Q000201:enzymology* / Q000502:physiology; D018613:Microscopy, Confocal; D020928:Mitogen-Activated Protein Kinases / Q000502:physiology; D051381:Rats; D044464:Receptor, PAR-2; D018179:Receptors, Thrombin / Q000502:physiology; D012697:Serine Endopeptidases / Q000097:blood / Q000502:physiology*; D013010:Sonication; D018808:Transcription Factor AP-1 / Q000097:blood / Q000502:physiology*; D053802:Tryptases
D016207:Cytokines; D015850:Interleukin-6; D016209:Interleukin-8; D044464:Receptor, PAR-2; D018179:Receptors, Thrombin; D018808:Transcription Factor AP-1; D020928:Mitogen-Activated Protein Kinases; D012697:Serine Endopeptidases; C509223:Tpsab1 protein, rat; C509221:Tpsb2 protein, rat; D053802:Tryptases