Summary
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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References
Aldenborg, F. & Enerbäck, L. (1988) Histochemical heterogeneity of dermal mast cells in athymic and normal rats. Histochem. J. 20, 19–28.
Benditt, E. P. & Arase, M. (1959) An enzyme in mast cells with properties like chymotrypsin. J. Exp. Med. 110, 451–60.
Bentley, A. M., Jacobson, M. R., Cumberworth, V., Barkans, J. R., Moqbel, R., Schwartz, L. B., Irani, A.-M. A., Kay, A. B. & Durham, S. R. (1992) Immunohistology of the nasal mucosa in seasonal allergic rhinitis: increases in activated eosinophils and epithelial mast cells. J. Allergy Clin. Immunol. 89, 877–83.
Blenkinsopp, W. K. (1967) Mast cell proliferation in adult rats. J. Cell Sci. 2, 33–7.
Castells, M. C., Irani, A. M. & Schwartz, L. B. (1987) Evaluation of human peripheral blood leukocytes for mast cell tryptase. J. Immunol. 138, 2184–9.
Caughey, G. H., Viro, N. F., Calonico, L. D., MacDonald, D. M., Lazarus, S. C. & Gold, W. M. (1988) Chymase and tryptase in dog mastocytoma cells: asynchronous expression as revealed by enzyme cytochemical staining. J. Histochem. Cytochem. 36, 1053–60.
Chiu, H. & Lagunoff, D. (1972) Histochemical comparison of vertebrate mast cells. Histochem. J. 4, 135–44.
Craig, S. S., Schechter, N. M. & Schwartz, L. B. (1989) Ultrastructural analysis of maturing human T and TC mast cells in situ. Lab. Invest. 60, 147–57.
Enerbäck, L. (1986) Mast cell heterogeneity: the evolution of the concept of a specific mucosal mast cell. In Mast Cell Differentiation and Heterogeneity (edited by Befus, A. D., Bienenstock, J. & Denburg, J. A.), pp. 1–26. New York: Raven Press.
Enerbäck, L. & Löwhagen, G. B. (1979) Long term increase of mucosal mast cells in the rat induced by administration of compound 48/80. Cell Tissue Res. 198, 209–15.
Enerbäck, L., Kolset, S. O., Kusche, M., Hjerpe, A. & Lindahl, U. (1985) Glycosaminoglycans in rat mucosal mast cells. Biochem. J. 227, 661–8.
Enerbäck, L., Miller, H. R. P. & Mayrhofer, G. (1986) Methods for the identification and characterisation of mast cells by light microscopy. In Mast Cell Differentiation and Heterogeneity (edited by Befus, A. D., Bienenstock, J. & Denburg, J. A.), pp. 405–17. New York: Raven Press.
Enerbäck, L., Pipkorn, U., Aldenborg, F. & Wingren, U. (1989) Mast cell heterogeneity in man: properties and function of human mucosal mast cells. In Mast Cell and Basophil Differentiation and Function in Health and Disease (edited by Galli, S. J. & Austen, K. F.), pp. 27–37. New York: Raven Press.
Gibson, S. & Miller, H. R. (1986) Mast cell subsets in the rat distinguished immunohistochemically by their content of serine proteinases. Immunology 58, 101–4.
Glenner, G. C. & Cohen, L. A. (1960) Histochemical demonstration of species-specific trypsin-like enzyme in mast cells. Nature 185, 846–7.
Huntley, J. F., Newlands, G. F., Gibson, S., Ferguson, A. & Miller, H. R. (1985) Histochemical demonstration of chymotrypsin like esterases in mucosal mast cells in four species including man. J. Clin. Pathol. 38, 375–84.
Irani, A.-M. A. & Schwartz, L. B. (1990) Neutral proteases as indicators of human mast cell heterogeneity. Monogr. Allergy 27, 146–62.
Irani, A.-M. A., Schechter, N. M., Craig, S. S., Deblois, G. & Schwartz, L. B. (1986) Two types of human mast cells that have distinct neutral protease compositions. Proc. Natl Acad. Sci. USA 83, 4464–8.
Irani, A.-M. A., Craig, S. S., Deblois, G., Elson, C. O., Schechter, N. M. & Schwartz, L. B. (1987) Deficiency of the tryptase-positive, chymase-negative mast cell type in gastrointestinal mucosa of patients with defective T lymphocyte function. J. Immunol. 138, 4381–6.
Irani, A.-M. A., Bradford, T. R., Kepley, C. L., Schechter, N. M. & Schwartz, L. B. (1989) Detection of MCT and MCTC types of human mast cells by immunohistochemistry using new monoclonal anti-tryptase and anti-chymase antibodies. J. Histochem. Cytochem. 37, 1509–15.
Juliusson, S., Aldenborg, F. & Enerbäck, L. (1994) Proteinase content of mast cells of the nasal mucosa; effects of natural allergen exposure and of local corticosteroid treatment. Allergy (in press).
King, S. J., Miller, H. R., Woodbury, R. G. & Newlands, G. F. (1986) Gut mucosal mast cells in Nippostrongylus -primed rats are the major source of secreted rat mast cell protease II following systemic anaphylaxis. Eur. J. Immunol. 16, 151–5.
Lagunoff, D. & Benditt, E. P. (1963) Proteolytic enzymes of mast cells. Ann. N.Y. Acad. Sci. 103, 185–98.
Larsson, L.-I. (1988) Immunocytochemistry: Theory and Practice. Boca Raton, FL: CRC Press Inc.
Matin, R., Tam, E. K., Nadel, J. A. & Caughey, G. H. (1992) Distribution of chymase-containing mast cells in human bronchi. J. Histochem. Cytochem. 40, 781–86.
Miller, J. S. & Schwartz, L. B. (1989) Human mast cell proteases and mast cell heterogeneity. Curr. Opin. Immunol. 1, 637–42.
Miller, J. S., Westin, E. H. & Schwartz, L. B. (1989) Cloning and characterization of complementary DNA for human tryptase. J. Clin. Invest. 84, 1188–95.
Miller, H. R., Huntley, J. F., Newlands, G. F. & Irvine, J. (1990) Granule chymases and the characterization of mast cell phenotype and function in rat and mouse. Monogr. Allergy 27, 1–30.
Miller, J. S., Moxley, G. & Schwartz, L. B. (1990) Cloning and characterization of a second complementary DNA for human tryptase. J. Clin. Invest. 86, 864–70.
Osman, I. A. R., Garrett, J. R. & Smith, R. E. (1989) Enzyme histochemical discrimination between tryptase and chymase in mast cells of human gut. J. Histochem. Cytochem. 4, 415–21.
Schechter, N. M., Slavin, D., Fetter, R. D., Lazarus, G. S. & Fräki, J. E. (1988) Purification and identification of two serine class proteinases from dog mast cells biochemically and immunologically similar to human proteinases tryptase and chymase. Arch. Biochem. Biophys. 262, 232–44.
Schwartz, L. B., Lewis, R. A. & Austen, K. F. (1981) Tryptase from human pulmonary mast cells. Purification and characterization. J. Biol. Chem. 256, 11 939–43.
Stevens, R. L., Lee, T. D., Seldin, D. C., Austen, K. F., Befus, A. D. & Bienenstock, J. (1986) Intestinal mucosal mast cells from rats infected with Nippostrongylus brasiliensis contain protease-resistant chondroitin sulfate di-B proteoglycans. J. Immunol. 137, 291–5.
Vanderslice, P., Ballinger, S. M., Tam, E. K., Goldstein, S. M., Craik, C. S. & Caughey, G. H. (1990) Human mast cell tryptase: multiple cDNAs and genes reveal a multigene serine protease family. Proc. Natl Acad. Sci. USA 87, 3811–5.
Walls, A. F., Jones, D. B., Williams, J. H., Church, M. K. & Holgate, S. T. (1990) Immunohistochemical identification of mast cells in formaldehyde-fixed tissue using monoclonal antibodies specific for tryptase. J. Pathol. 162, 119–26.
Wintroub, B. U., Kaempfer, C. E., Schechter, N. M. & Proud, D. (1986) A human lung mast cell chymotrypsin-like enzyme. Identification and partial characterization. J. Clin. Invest. 77, 196–201.
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Aldenborg, F., Enerbäck, L. The immunohistochemical demonstration of chymase and tryptase in human intestinal mast cells. Histochem J 26, 587–596 (1994). https://doi.org/10.1007/BF00158593
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DOI: https://doi.org/10.1007/BF00158593