Mast cell tryptase and chymase in chronic leg ulcers: chymase is potentially destructive to epithelium and is controlled by proteinase inhibitors
M. Huttunen,
I.T. Harvima,
M. Huttunen
Department of Dermatology, Kuopio University Hospital, POB 1777, 70211 Kuopio, Finland
Department of Dermatology, Helsinki University Central Hospital, Helsinki, Finland
Search for more papers by this authorI.T. Harvima
Department of Dermatology, Kuopio University Hospital, POB 1777, 70211 Kuopio, Finland
Search for more papers by this authorM. Huttunen,
I.T. Harvima,
M. Huttunen
Department of Dermatology, Kuopio University Hospital, POB 1777, 70211 Kuopio, Finland
Department of Dermatology, Helsinki University Central Hospital, Helsinki, Finland
Search for more papers by this authorI.T. Harvima
Department of Dermatology, Kuopio University Hospital, POB 1777, 70211 Kuopio, Finland
Search for more papers by this authorIlkka T. Harvima.
E-mail: [email protected]
Conflicts of interest: none declared.
Summary
Background Numerous mast cells are present in chronic leg ulcers. Tryptase and chymase are the major mediators of mast cells, but their significance is mostly dependent on their activity. In addition, the proteinases may affect ulcer epithelialization.
Objectives To study levels and activity of tryptase and chymase in wash samples and biopsies from chronic leg ulcers and the possible effect of these proteinases on keratinocyte growth and adherence.
Methods Wash samples were taken from 16 patients and a superficial shave biopsy was taken in eight of these patients; a second biopsy series was obtained from the edge of chronic venous leg ulcers (n = 6).
Results Significant levels of soluble tryptase activity and histamine, but low levels of chymase activity, were measured in wash samples from chronic ulcers. No tryptase-inhibiting activity, but clear chymase-inhibiting activity, was detected in the wash samples. In superficial wound bed biopsies, relatively marked levels of chymase activity together with histamine and tryptase activity were detected. In the second biopsy series, about 80% of the mast cells belonged to the MCTC type (tryptase- and chymase-immunopositive). However, about 55–61% of the chymase-immunopositive cells displayed chymase activity and 64 ± 17% of the tryptase-positive cells revealed immunoreactivity of α1-antichymotrypsin. As the activity of chymase and tryptase was detected in the ulcer base in a ratio of 1 : 8, a preparation containing both chymase and tryptase was partially purified from human skin yielding a similar activity ratio of 1 : 11–13. Treatment of fibronectin-coated plastic surfaces with this preparation decreased the adherence of cultured human keratinocytes, this effect being attributable mainly to chymase. In 2-day cultures using growth factor/serum-deficient low- or high-calcium medium, the tryptase–chymase preparation inhibited the slow growth and at higher concentrations it even induced detachment of keratinocytes. This effect was attributed to chymase, and it was partially regulated by heparin and histamine.
Conclusions Even though chymase is partially inactivated in chronic leg ulcers, accumulated mast cells in the close proximity of the epithelium edge and their chymase may impair keratinocyte adherence and migration.
References
- 1 Briggaman RA, Schechter NM, Fräki JE, Lazarus GS. Degradation of the epidermal–dermal junction by proteolytic enzymes from human skin and human polymorphonuclear leukocytes. J Exp Med 1984; 160: 1027–42.
- 2 Kaminska R, Naukkarinen A, Glinski W et al. Mast cells in developing subepidermal bullous diseases: emphasis on tryptase, chymase and protease inhibitors. Acta Derm Venereol (Stockh) 1999; 79: 351–5.DOI: 10.1080/000155599750010247
- 3 D'Auria L, Pietravalle M, Cordiali-Fei P, Ameglio F. Increased tryptase and myeloperoxidase levels in blister fluids of patients with bullous pemphigoid: correlations with cytokines, adhesion molecules and anti-basement membrane zone antibodies. Exp Dermatol 2000; 9: 131–7.DOI: 10.1034/j.1600-0625.2000.009002131.x
- 4 Green RM, Cordero A, Winkelmann RK. Epidermal mast cells. Arch Dermatol 1977; 113: 166–9.DOI: 10.1001/archderm.113.2.166
- 5 Bolton LL, Montagna W. Mast cells in human ulcers. Am J Dermatopathol 1993; 15: 133–8.
- 6 Harvima IT, Naukkarinen A, Paukkonen K et al. Mast cell tryptase and chymase in developing and mature psoriatic lesions. Arch Dermatol Res 1993; 285: 184–92.DOI: 10.1007/BF00372007
- 7 Huttunen M, Aalto M-L, Harvima RJ et al. Alterations in mast cells showing tryptase and chymase activity in epithelializating and chronic wounds. Exp Dermatol 2000; 9: 258–65.DOI: 10.1034/j.1600-0625.2000.009004258.x
- 8 Ackermann L, Harvima IT. Mast cells of psoriatic and atopic dermatitis skin are positive for TNF-α and their degranulation is associated with expression of ICAM-1 in the epidermis. Arch Dermatol Res 1998; 290: 353–9.DOI: 10.1007/s004030050317
- 9 Symington FW. Lymphotoxin, tumor necrosis factor, and gamma interferon are cytostatic for normal human keratinocytes. J Invest Dermatol 1989; 92: 798–805.DOI: 10.1111/1523-1747.ep12696816
- 10 Pillai S, Bikle DD, Eessalu TE et al. Binding and biological effects of tumor necrosis factor alpha on cultured human neonatal foreskin keratinocytes. J Clin Invest 1989; 83: 816–21.
- 11 Pillai S, Gilliam L, Conrad HE, Holleran WM. Heparin and its non-anticoagulant analogues inhibit human keratinocyte growth without inducing differentiation. J Invest Dermatol 1994; 103: 647–50.DOI: 10.1111/1523-1747.ep12398386
- 12 Huttunen M, Hyttinen M, Nilsson G et al. Inhibition of keratinocyte growth in cell culture and whole skin culture by mast cell mediators. Exp Dermatol 2001; 10: 184–92.DOI: 10.1034/j.1600-0625.2001.010003184.x
- 13 Imaizumi T, Jean-Louis F, Dubertret M-L, Dubertret L. Heparin induces fibroblast proliferation, cell–matrix interaction and epidermal growth inhibition. Exp Dermatol 1996; 5: 89–95.
- 14 Irani A-MA, Bradford TR, Kepley CL et al. Detection of MCT and MCTC types of human mast cells by immunohistochemistry using new monoclonal anti-tryptase and anti-chymase antibodies. J Histochem Cytochem 1989; 37: 1509–15.
- 15 Weidner N, Austen KF. Heterogeneity of mast cells at multiple body sites: fluorescent determination of avidin binding and immunofluorescent determination of chymase, tryptase, and carboxypeptidase content. Pathol Res Pract 1993; 189: 156–62.
- 16 Harvima RJ, Schwartz LB. Mast cell-derived mediators. In: Immunopharmacology of Mast Cells and Basophils ( J Foreman, ed.). London: Academic Press, 1993; 115–38.
- 17 Gibbs BF, Wierecky J, Welker P et al. Human skin mast cells rapidly release preformed and newly generated TNF-α and IL-8 following stimulation with anti-IgE and other secretagogues. Exp Dermatol 2001; 10: 312–20.DOI: 10.1034/j.1600-0625.2001.100503.x
- 18 Kivinen PK, Kaminska R, Naukkarinen A et al. Release of soluble tryptase but only minor amounts of chymase activity from cutaneous mast cells. Exp Dermatol 2001; 10: 246–55.DOI: 10.1034/j.1600-0625.2001.100404.x
- 19 Kivinen PK, Nilsson G, Naukkarinen A, Harvima IT. Mast cell survival and apoptosis in organ-cultured human skin. Exp Dermatol 2003; 12: 53–60.DOI: 10.1034/j.1600-0625.2003.120107.x
- 20 Kaminska R, Helisalmi P, Harvima RJ et al. Focal dermal–epidermal separation and fibronectin cleavage in basement membrane by human mast cell tryptase. J Invest Dermatol 1999; 113: 567–73.DOI: 10.1046/j.1523-1747.1999.00738.x
- 21 Lohi J, Harvima I, Keski-Oja J. Pericellular substrates of human mast cell tryptase: 72,000 dalton gelatinase and fibronectin. J Cell Biochem 1992; 50: 337–49.DOI: 10.1002/jcb.240500402
- 22 Lazaar AL, Plotnick MI, Kucich U et al. Mast cell chymase modifies cell–matrix interactions and inhibits mitogen-induced proliferation of human airway smooth muscle cells. J Immunol 2002; 169: 1014–20.
- 23 Saarinen JV, Kalkkinen N, Welgus HG, Kovanen PT. Activation of human interstitial procollagenase through direct cleavage of the Leu83-Thr84 bond by mast cell chymase. J Biol Chem 1994; 269: 18134–40.
- 24 Gruber BL, Marchese MJ, Suzuki K et al. Synovial procollagenase activation by human mast cell tryptase: dependence upon matrix metalloproteinase 3 activation. J Clin Invest 1989; 84: 1657–62.
- 25 Stack MS, Johnson DA. Human mast cell tryptase activates single-chain urinary-type plasminogen activator (pro-urokinase). J Biol Chem 1994; 269: 9416–19.
- 26 Abe M, Kurosawa M, Ishikawa O et al. Mast cell tryptase stimulates both human dermal fibroblast proliferation and type I collagen production. Clin Exp Allergy 1998; 28: 1509–17.DOI: 10.1046/j.1365-2222.1998.00360.x
- 27 Cairns JA, Walls AF. Mast cell tryptase stimulates the synthesis of type I collagen in human lung fibroblasts. J Clin Invest 1997; 99: 1313–21.
- 28 Blair RJ, Meng H, Marchese MJ et al. Human mast cells stimulate vascular tube formation: tryptase is a novel, potent angiogenic factor. J Clin Invest 1997; 99: 2691–700.
- 29 Kofford MW, Schwartz LB, Schechter NM et al. Cleavage of type I procollagen by human mast cell chymase initiates collagen fibril formation and generates a unique carboxyl-terminal propeptide. J Biol Chem 1997; 272: 7127–31.DOI: 10.1074/jbc.272.11.7127
- 30 Taipale J, Lohi J, Saarinen JV et al. Human mast cell chymase and leukocyte elastase release latent transforming growth factor-β1 from the extracellular matrix of cultured human epithelial and endothelial cells. J Biol Chem 1995; 270: 4689–96.DOI: 10.1074/jbc.270.9.4689
- 31 Jordana M, Befus AD, Newhouse MT et al. Effect of histamine on proliferation of normal human adult lung fibroblasts. Thorax 1988; 43: 552–8.
- 32 Hatamochi A, Ueki H, Mauch C, Krieg T. Effect of histamine on collagen and collagen m-RNA production in human skin fibroblasts. J Dermatol Sci 1991; 2: 407–12.
- 33 Artuc M, Hermes B, Steckelings UM et al. Mast cells and their mediators in cutaneous wound healing—active participants or innocent bystanders. Exp Dermatol 1999; 8: 1–16.DOI: 10.1159/000026062
- 34 Rice KD, Tanaka RD, Katz BA et al. Inhibitors of tryptase for the treatment of mast cell-mediated diseases. Current Pharm Design 1998; 4: 381–96.
- 35 Schechter NM, Sprows JL, Schoenberger OL et al. Reaction of human skin chymotrypsin-like proteinase chymase with plasma proteinase inhibitors. J Biol Chem 1989; 264: 21308–15.
- 36 Walter M, Sutton RM, Schechter NM. Highly efficient inhibition of human chymase by α(2)-macroglobulin. Arch Biochem Biophys 1999; 368: 276–84.DOI: 10.1006/abbi.1999.1309
- 37 Grinnell F, Zhu M. Fibronectin degradation in chronic wounds depends on the relative levels of elastase, α1-proteinase inhibitor, and α2-macroglobulin. J Invest Dermatol 1996; 106: 335–41.DOI: 10.1111/1523-1747.ep12342990
- 38 Harvima IT, Harvima RJ, Penttilä IM et al. Effect of human mast cell tryptase on human plasma proenzymes. Int Arch Allergy Appl Immunol 1989; 90: 104–8.
- 39 Harvima RJ, Harvima IT, Fräki JE. Optimization of histamine radio enzyme assay with purified histamine-N-methyltransferase. Clin Chim Acta 1988; 171: 247–56.DOI: 10.1016/0009-8981(88)90150-7
- 40 Harvima IT, Schechter NM, Harvima RJ, Fräki JE. Human skin tryptase: purification, partial characterization and comparison with human lung tryptase. Biochim Biophys Acta 1988; 957: 71–80.
- 41 Harvima IT, Lappalainen K, Hirvonen M-R et al. Heparin modulates the growth and adherence and augments the growth-inhibitory action of TNF-α on cultured human keratinocytes. J Cell Biochem 2004; 92: 372–86.DOI: 10.1002/jcb.20068
- 42 Otto W. Determination of cell number by DNA content. In: Keratinocyte Handbook ( IM Leigh, FM Watt, eds). Cambridge: Cambridge University Press, 1994; 89–90.
- 43 Schechter NM, Irani A-MA, Sprows JL et al. Identification of a cathepsin G-like proteinase in the MCTC type of human mast cells. J Immunol 1990; 145: 2652–61.
- 44 Schechter NM, Fräki JE, Geesin JC, Lazarus GS. Human skin chymotryptic proteinase: isolation and relation to cathepsin G and rat mast cell proteinase I. J Biol Chem 1983; 258: 2973–8.
- 45 Eggli PS, Graber W. Cytochemical localization of hyaluronan in rat and human skin mast cell granules. J Invest Dermatol 1993; 100: 121–5.DOI: 10.1111/1523-1747.ep12462777
- 46 Schwartz LB, Atkins PC, Bradford TR et al. Release of tryptase together with histamine during the immediate cutaneous response to allergen. J Allergy Clin Immunol 1987; 80: 850–5.
- 47 Kaminska R, Naukkarinen A, Horsmanheimo M, Harvima IT. Suction blister formation in skin after acute and repeated mast cell degranulation. Acta Derm Venereol (Stockh) 1999; 79: 191–4.DOI: 10.1080/000155599750010940
- 48 Saarinen JV, Harvima RJ, Naukkarinen A et al. The release of histamine is associated with the inactivation of mast cell chymase during immediate allergic wheal reaction in the skin. Clin Exp Allergy 2001; 31: 593–601.DOI: 10.1046/j.1365-2222.2001.01030.x
- 49 Algermissen B, Hermes B, Feldmann-Boeddeker I et al. Mast cell chymase and tryptase during tissue turnover: analysis on in vitro mitogenesis of fibroblasts and keratinocytes and alterations in cutaneous scars. Exp Dermatol 1999; 8: 193–8.DOI: 10.1159/000014590
- 50
Schechter NM,
Brass LF,
Lavker RM,
Jensen PJ.
Reaction of mast cell proteases tryptase and chymase with protease activated receptors (PARs) on keratinocytes and fibroblasts.
J Cell Physiol
1998; 176: 365–73.DOI: 10.1002/(SICI)1097-4652(199808)176:2<365::AID-JCP15>3.0.CO;2-2
10.1002/(SICI)1097-4652(199808)176:2<365::AID-JCP15>3.0.CO;2-2 CAS PubMed Web of Science® Google Scholar
- 51 Steinhoff M, Corvera CU, Thoma MS et al. Proteinase-activated receptor-2 in human skin: tissue distribution and activation of keratinocytes by mast cell tryptase. Exp Dermatol 1999; 8: 282–94.
- 52 Palolahti M, Lauharanta J, Stephens RW et al. Proteolytic activity in leg ulcer exudate. Exp Dermatol 1993; 2: 29–37.
- 53 Saarialho-Kere UK. Patterns of matrix metalloproteinases and TIMP expression in chronic ulcers. Arch Dermatol Res 1998; 290 (Suppl.): S47–54.
- 54 Wysocki AB, Staiano-Coico L, Grinnell F. Wound fluid from chronic leg ulcers contains elevated levels of metalloproteinases MMP-2 and MMP-9. J Invest Dermatol 1993; 101: 64–8.DOI: 10.1111/1523-1747.ep12359590
- 55 Barbosa Pereira PJ, Bergner A, Macedo-Ribeiro S et al. Human β-tryptase is a ring-like tetramer with active sites facing a central pore. Nature 1998; 392: 306–10.
- 56 Xu X, Rivkind A, Pappo O et al. Role of mast cells and myofibroblasts in human peritoneal adhesion formation. Ann Surg 2002; 236: 593–601.DOI: 10.1097/00000658-200211000-00009
- 57 McEuen AR, Sharma B, Walls AF. Regulation of the activity of human chymase during storage and release from mast cells: the contributions of inorganic cations, pH, heparin and histamine. Biochim Biophys Acta 1995; 1267: 115–21.DOI: 10.1016/0167-4889(95)00066-2
- 58 Walter M, Plotnick M, Schechter NM. Inhibition of human mast cell chymase by secretory leukocyte proteinase inhibitor: enhancement of the interaction by heparin. Arch Biochem Biophys 1996; 327: 81–8.DOI: 10.1006/abbi.1996.0095
