Specific inhibition of beta-tryptase expression in a human mast cell line by granulocyte-macrophage colony-stimulating factor produced by airways structural cells.
The growth and differentiation of mast cells are regulated by cytokines produced in tissue microenvironments. We previously reported that mast cells isolated from the epithelial compartment of nasal polyp tissue contain significantly less tryptase when compared with mast cells isolated from the stroma of the same tissue. In an attempt to explore this finding, we analyzed the ability of supernatants obtained from cultured nasal polyp epithelial cells (NP-EpCM) or nasal polyp fibroblasts (NP-FbCM) to regulate the tryptase content of the immature human mast cell line HMC-1. HMC-1 cells were cultured for 7 days in Iscove's modified Dulbecco's medium (IMDM) with 30% of either NP-FbCM or NP-EpCM or 20% MoCM (supernatant of a leukemic T cell line). As assessed by radioimmunoassay and test for enzymatic activity, all three conditioned media were shown to significantly decrease tryptase protein expression in HMC-1, when compared with cultures performed with IMDM alone (NP-EpCM P < 0.001; NP-FbCM P < 0.04; MoCM P < 0.004). In addition, Northern blot analysis demonstrated lower tryptase mRNA levels upon exposure to all three conditioned media tested, suggesting that tryptase downregulation occurs at the transcriptional level. In further studies we found that preincubation of MoCM with anti-granulocyte-macrophage colony-stimulating factor (GM-CSF) completely blocked the observed downregulation of tryptase expression mediated by this conditioned medium. The findings suggest that GM-CSF has a suppressive effect on expression of protease in mast cells, and may thus play a modulatory role in determining the extent of tissue inflammation in allergic airways disease.
Finotto
S
S
Department of Medicine, McMaster University, Hamilton, Ontario, Canada.
Marshall
J S
JS
Butterfield
J H
JH
Denburg
J A
JA
D015152:Blotting, Northern; D002455:Cell Division / Q000187:drug effects / Q000276:immunology; D053818:Chymases; D017077:Culture Media, Conditioned / Q000494:pharmacology; D010450:Endopeptidases / Q000378:metabolism; D005347:Fibroblasts / Q000737:chemistry; D015870:Gene Expression / Q000187:drug effects; D016178:Granulocyte-Macrophage Colony-Stimulating Factor / Q000494:pharmacology*; D006632:Histamine / Q000378:metabolism; D006801:Humans; D007150:Immunohistochemistry; D018836:Inflammation Mediators / Q000502:physiology*; D015458:Leukemia, T-Cell; D008168:Lung / Q000166:cytology* / Q000276:immunology; D008407:Mast Cells / Q000166:cytology / Q000187:drug effects* / Q000201:enzymology; D009298:Nasal Polyps; D012333:RNA, Messenger / Q000378:metabolism; D012697:Serine Endopeptidases / Q000096:biosynthesis / Q000235:genetics*; D053802:Tryptases; D014407:Tumor Cells, Cultured / Q000166:cytology / Q000187:drug effects / Q000201:enzymology; D014409:Tumor Necrosis Factor-alpha / Q000378:metabolism
D017077:Culture Media, Conditioned; D018836:Inflammation Mediators; D012333:RNA, Messenger; D014409:Tumor Necrosis Factor-alpha; D006632:Histamine; D016178:Granulocyte-Macrophage Colony-Stimulating Factor; D010450:Endopeptidases; D012697:Serine Endopeptidases; C083294:chymase 2; D053818:Chymases; D053802:Tryptases