Tryptase, the dominant secretory granular protein in human mast cells, is a potent mitogen for cultured dog tracheal smooth muscle cells.
Hyperplasia of airway smooth muscle cells is present in the airways of asthmatic patients and may contribute to the development of the bronchial hyperresponsiveness that occurs in these patients. Because tryptase is an abundant component of mast cell granules and has demonstrated growth-stimulatory effects in other mesenchymal cells (J. Clin. Invest. 1991; 88:493-499), the goal of our study was to determine whether tryptase is a mitogen for airway smooth muscle cells. The mitogenic effects of tryptase were tested in passages 1 through 5 of dog tracheal smooth muscle cells, either by counting smooth muscle cells or by monitoring uptake of bromodeoxyuridine (BrdU) into cellular DNA during S-phase. With respect to its efficacy, at a near maximal concentration (4 nM), tryptase increased cell numbers 2.1 +/- 0.2- or 2.8 +/- 0.6-fold above controls after 2 or 4 days, respectively, and these increases were approximately the same as those induced by platelet-derived growth factor (50 ng/ml) or 10% calf serum. With respect to potency, tryptase caused concentration-dependent increases in BrdU uptake, as detected in an enzyme-linked immunosorbent assay or by counting BrdU-labeled nuclei, with an EC50 of 2 nM. Pretreatment of tryptase with diisopropylfluorophosphate, to reduce markedly its catalytic as a activity as a proteinase, attenuated its growth-stimulated effects by 58 +/- 16%. Tryptase-induced mitogenesis was not a nonspecific effect of all serine proteinases, because thrombin, another proteinase with mitogenicity for fibroblasts, stimulated neither increases in cell counts nor BrdU uptake in our cells. We conclude that tryptase is a potent mitogen for airway smooth muscle cells in culture.
Brown
J K
JK
Pulmonary and Critical Care Medicine Section, VA Medical Center, San Francisco, CA 94121, USA.
Tyler
C L
CL
Jones
C A
CA
Ruoss
S J
SJ
Hartmann
T
T
Caughey
G H
GH
D000818:Animals; D001973:Bromodeoxyuridine; D002452:Cell Count; D002455:Cell Division / Q000187:drug effects; D002478:Cells, Cultured / Q000502:physiology; D053818:Chymases; D003594:Cytoplasmic Granules / Q000201:enzymology*; D004247:DNA / Q000096:biosynthesis; D004285:Dogs; D004305:Dose-Response Relationship, Drug; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D007150:Immunohistochemistry; D018836:Inflammation Mediators / Q000494:pharmacology*; D007531:Isoflurophate / Q000494:pharmacology; D008407:Mast Cells / Q000201:enzymology / Q000502:physiology*; D008934:Mitogens / Q000502:physiology*; D009130:Muscle, Smooth / Q000166:cytology* / Q000378:metabolism; D012697:Serine Endopeptidases / Q000494:pharmacology*; D014132:Trachea / Q000166:cytology / Q000378:metabolism; D053802:Tryptases
D018836:Inflammation Mediators; D008934:Mitogens; D007531:Isoflurophate; D004247:DNA; D012697:Serine Endopeptidases; C083294:chymase 2; D053818:Chymases; D053802:Tryptases; D001973:Bromodeoxyuridine