A novel function for transforming growth factor-beta1: upregulation of the expression and the IgE-independent extracellular release of a mucosal mast cell granule-specific beta-chymase, mouse mast cell protease-1.
Intestinal mucosal mast cells (IMMC) express granule neutral proteases that are regulated by T-cell-derived cytokines, including interleukin-3 (IL-3) and IL-9, and by stem cell factor (SCF). The IMMC-specific chymase, mouse mast cell protease-1 (mMCP-1), is released in substantial quantities into the blood stream during gastrointestinal allergic responses. We used cultured bone marrow-derived mast cells (mBMMC) to identify cytokines that regulate the expression and extracellular release of mMCP-1. When grown in IL-3-rich WEHI (15% vol/vol) and 50 ng/mL recombinant rat SCF (rrSCF) bone marrow cells supplemented with IL-9 (5 ng/mL) differentiated into mBMMC that expressed a maximum of less than 250 ng mMCP-1/10(6) cells and 189 ng mMCP-1/mL of culture supernatant. Supplementation of the same three cytokines with transforming growth factor-beta1 (TGF-beta1; 1 ng/mL) resulted in substantially enhanced expression (6 micrograms/10(6) mBMMC) and extracellular release (2 micrograms/mL of culture supernatant) of mMCP-1. The response to TGF-beta1 was dose-dependent, with maximal effect at 1 ng/mL, and was associated with immunohistochemical and ultrastructural changes in the secretory granules. IL-9-induced expression of mMCP-1 may be due to endogenously expressed TGF-beta1, because it was blocked by anti-TGF-beta antibodies. In conclusion, the expression and extracellular release of the IMMC-specific chymase, mMCP-1, is strictly regulated by TGF-beta1.
Miller
H R
HR
Department of Veterinary Clinical Studies, Royal (Dick) School of Veterinary Studies, The University of Edinburgh, Easter Bush Veterinary Centre, Easter Bush, Roslin, Midlothian, Scotland. Hugh.Miller@ed.ac.uk
Wright
S H
SH
Knight
P A
PA
Thornton
E M
EM
D000818:Animals; D000906:Antibodies / Q000494:pharmacology; D001854:Bone Marrow Cells / Q000166:cytology; D002478:Cells, Cultured; D053818:Chymases; D017077:Culture Media, Conditioned; D016207:Cytokines / Q000494:pharmacology / Q000502:physiology*; D015971:Gene Expression Regulation, Enzymologic / Q000276:immunology*; D007073:Immunoglobulin E / Q000494:pharmacology / Q000502:physiology*; D007377:Interleukin-3 / Q000494:pharmacology; D016906:Interleukin-9 / Q000494:pharmacology; D007700:Kinetics; D008297:Male; D008407:Mast Cells / Q000166:cytology / Q000201:enzymology / Q000276:immunology*; D051379:Mice; D008807:Mice, Inbred BALB C; D051381:Rats; D011994:Recombinant Proteins / Q000494:pharmacology; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012697:Serine Endopeptidases / Q000235:genetics* / Q000378:metabolism*; D019089:Stem Cell Factor / Q000494:pharmacology; D014158:Transcription, Genetic; D016212:Transforming Growth Factor beta / Q000494:pharmacology / Q000502:physiology*; D014407:Tumor Cells, Cultured
D000906:Antibodies; D017077:Culture Media, Conditioned; D016207:Cytokines; D007377:Interleukin-3; D016906:Interleukin-9; D011994:Recombinant Proteins; D019089:Stem Cell Factor; D016212:Transforming Growth Factor beta; D007073:Immunoglobulin E; D012697:Serine Endopeptidases; D053818:Chymases; C509230:Mcpt1 protein, mouse; C509231:Mcpt1 protein, rat