Purification and identification of two serine class proteinases from dog mast cells biochemically and immunologically similar to human proteinases tryptase and chymase

Schechter, Norman M.; Slavin, Dorothy; Fetter, Richard D.; Lazarus, Gerald S.; Fräki, Jorma E.

doi:10.1016/0003-9861(88)90185-3

Cited by 45 publications

(26 citation statements)

References 33 publications

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“…Interspecies antigen cross-reactivity between the human and dog masbcell proteinases was previously found using polyclonal antibodies (Schechter et al, 1988).…”

Section: Distribution Of Proteinase-containing Mast Cellsmentioning

confidence: 90%

The immunohistochemical demonstration of chymase and tryptase in human intestinal mast cells

Aldenborg

Enerbäck

1994

Histochem J

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An immunohistochemical double-labelling technique for the simultaneous identification of mast cells containing tryptase alone (MCT) or chymase together with tryptase (MCTC) was evaluated quantitatively using two monoclonal antibodies, mAb 1222A (antitryptase) and mAb 1254B (antichymase). Saturation conditions were established for the binding of the antibodies to the mast cell enzymes by counting labelled mast cells in consecutive sections of normal human intestine incubated with serial dilutions of the antibodies. When, under such conditions, the antitryptase was applied after saturation with mAb 1254B, the reproducibility of the double-labelling procedure was excellent. MCT were located preferentially in the intestinal mucosa but, in contrast to what has previously been reported, they were not the predominant type of mast cell at this site. The percentage of MCT of the total number of immunopositive mast cells varied considerably in the colonic mucosa (7-67%, average 30%), while this was not the case in the small intestinal mucosa (5-26%, average 10%). Mast cell chymase, unlike tryptase, was not recognized by the antichymase antibody after aldehyde fixation and a higher apparent fraction of MCT therefore occurred after double labelling. These findings suggest that the proteinase composition of human mast cells, unlike that of murine mast cells, should not be taken as evidence of phenotypic heterogeneity. Taken together with previous observations, they suggest instead that the lack of chymase may be related to functional activity or stage of maturation of the mast cells.

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“…Interspecies antigen cross-reactivity between the human and dog masbcell proteinases was previously found using polyclonal antibodies (Schechter et al, 1988).…”

Section: Distribution Of Proteinase-containing Mast Cellsmentioning

confidence: 90%

The immunohistochemical demonstration of chymase and tryptase in human intestinal mast cells

Aldenborg

Enerbäck

1994

Histochem J

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“…Human chymase was purified from skin as described (32,33). Its concentration was estimated from its specific activity for hydrolysis of the substrate Suc-Ala-Ala-Pro-Phe-4-nitroanilide (32)(33)(34). rHuSCF 165 (100 ng) was digested in 0.05 M Tris⅐HCl (pH 8.0) and 0.26 M NaCl with 100 nM human chymase.…”

Section: Methodsmentioning

confidence: 99%

Chymase cleavage of stem cell factor yields a bioactive, soluble product

Longley

Tyrrell

et al. 1997

Proc. Natl. Acad. Sci. U.S.A.

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171

106

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Stem cell factor (SCF) is produced by stromal cells as a membrane-bound molecule, which may be proteolytically cleaved at a site close to the membrane to produce a soluble bioactive form. The proteases producing this cleavage are unknown. In this study, we demonstrate that human mast cell chymase, a chymotrypsin-like protease, cleaves SCF at a novel site. Cleavage is at the peptide bond between Phe-158 and Met-159, which are encoded by exon 6 of the SCF gene. This cleavage results in a soluble bioactive product that is 7 amino acids shorter at the C terminus than previously identified soluble SCF. This research shows the identification of a physiologically relevant enzyme that specifically cleaves SCF. Because mast cells express the KIT protein, the receptor for SCF, and respond to SCF by proliferation and degranulation, this observation identifies a possible feedback loop in which chymase released from mast cell secretory granules may solubilize SCF bound to the membrane of surrounding stromal cells. The liberated soluble SCF may in turn stimulate mast cell proliferation and differentiated functions; this loop could contribute to abnormal accumulations of mast cells in the skin and hyperpigmentation at sites of chronic cutaneous inf lammation.

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“…The supernatants were used as the extract containing chymase (20). The enzyme activities of chymase were determined using the substrate 0.2 mM N-succinyl-Ala-Ala-Pro-Phe-p-nitroanilide (Sigma) containing 50 μg/ml heparin, 1 mg/ml bovine serum albumin, 1 mg/ml aprotinin, 2 M KCl and 100 mM Tris-HCl, pH 7.6 (21). In all assays, 50 μl of sample was added to a total reaction volume of 200 μl in a 96-well plate.…”

Section: Disease Activity Index (Dai)mentioning

confidence: 99%

Suppressive effect of non-anaphylactogenic anti-IgE antibody on the development of dextran sulfate sodium-induced colitis

Kang

Kim²,

Choi³

et al. 2006

Int J Mol Med

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Abstract. Inflammatory bowel disease (IBD) is a spectrum of immune-mediated chronic disorders of the intestine. Patients with IBD tend to exhibit significantly elevated levels of IgE in their serum. In general, the pathogenesis of IBD exhibits inflammatory events such as immunoglobulin E (IgE)-mediated hypersensitivity. We examined the effect of the non-anaphylactogenic anti-IgE antibody, which has been known to block IgE functions, in an animal model of ulcerative colitis induced by the oral intake of dextran sulfate sodium (DSS) for seven days. The non-anaphylactogenic anti-IgE antibody was subcutaneously injected on day 0 of DSS treatment. The disease activity index (DAI) was calculated by scoring intestinal states, including body weight loss, diarrhea, and rectal bleeding, and the activities of myeloperoxidase (MPO) and chymase were measured in the colon tissue. In addition, the expression of tumor necrosis factor (TNF)-· and cyclooxygenase (COX)-2 was determined by Western blotting. Administration of the anti-IgE antibody markedly reduced the histological damage to the colon and the DAI increment exhibited by the DSS-induced colitis. The anti-IgE antibody also significantly suppressed the activities of MPO and chymase as well as the expression of TNF-· and COX-2 in the DSS-treated colon tissue. Furthermore, the elevation of IgE levels in serum was induced by DSS and reduced by anti-IgE antibody injection. Thus, these results indicate that the IgE response played an important role in the clinical signs and the expression of inflammatory mediators in a colitis model caused by DSS treatment, suggesting that the non-anaphylactogenic anti-IgE antibody may be a useful therapeutic agent for ulcerative colitis.

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Purification and identification of two serine class proteinases from dog mast cells biochemically and immunologically similar to human proteinases tryptase and chymase

Cited by 45 publications

References 33 publications

The immunohistochemical demonstration of chymase and tryptase in human intestinal mast cells

The immunohistochemical demonstration of chymase and tryptase in human intestinal mast cells

Chymase cleavage of stem cell factor yields a bioactive, soluble product

Suppressive effect of non-anaphylactogenic anti-IgE antibody on the development of dextran sulfate sodium-induced colitis

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