Abstract
Mast cells generate mediators of inflammation which are stored in granules and secreted on activation either by allergen crosslinking of membrane-bound IgE or through other stimuli. Most methods for mast cell identification rely on the histochemical detection of constituents of the secretory granules. Although staining for mast cells with histochemical stains can be rapid and relatively inexpensive, it is not always possible to distinguish reliably between mast cells and basophils in tissues. A further problem with the staining of mast cells with commonly used basic dyes is that the reagents employed to fix the tissues can influence the results, leading to confusion regarding the numbers of mast cells present in various tissues. Recognition that there is considerable heterogeneity between mast cell populations in the degree to which staining properties are lost with formalin fixation has led to mast cell subsets being defined on this basis.
The development and application of procedures for identifying mast cell proteases has led to important advances in our understanding of the role of mast cells and in the nature of heterogeneity in man. The techniques described here should allow the reliable detection of mast cells and mast cell subsets in a range of tissues and cell preparations. There will be a continuing need for validation, for consideration of potential sources of error, and for the development of new and more reliable techniques for mast cell identification.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Similar content being viewed by others
References
Ehrlich, P. (1879) Contributions to the theory and practice of histological staining. In The Collected Papers of Paul Ehrlich (Himmelweit, F. ed.), 1956, Pergamon, New York, pp. 65–68.
Irani, A.-M. A., Bradford, T. R., Kepley, C. L., Schechter, N. M., and 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, 1509–1515.
Irani, A.-M. A., Bradford, T. R., and Schwartz, L. B. (1991) Human mast cell carboxypeptidase. Selective localization to MCTC cells. J. Immunol. 147, 247–253.
Walls, A. F., Roberts, J. A., Godfrey, R. C., Church, M. K., and Holgate, S. T. (1990) Histochemical heterogeneity of human mast cells: disease-related differences in mast cell subsets recovered by bronchoalveolar lavage. Int. Arch. Allergy Appl. Immunol. 92, 233–241.
Wingren, U. and Enerback, L. (1983) Mucosal mast cells of the rat intestine: a re-evaluation of fixation and staining properties, with special reference to protein blocking and solubility of the granular glycosaminoglycan. Histochemistry 15, 571–582.
Kimura, I., Moritani, Y., and Tanizaki, Y. (1973) Basophils in bronchial asthma with reference to reagin-type allergy. Clin. Allergy 3, 195–202.
Heard, B. E. (1986) Histochemical aspects of the staining of mast cells with particular reference to heterogeneity and quantification. In Asthma, Clinical Pharmacology and Therapeutic Progress (Kay, A. B. ed.) Blackwell Scientific Publications, Oxford, pp. 286–294.
Johansson, O., Virtanen, M., Hilliges, M., and Yang, Q. (1994) Histamine immunohistochemistry is superior to the conventional heparin-based routine staining methodology for investigations of human skin mast cells. Histochem. J. 26, 424–430.
Walls, A. F., Jones, D. B., Williams, J. H., Church, M. K., and Holgate, S. T. (1990) Immunohistochemical identification of mast cells in formaldehyde-fixed tissue using monoclonal antibodies specific for tryptase. J. Pathol. 162, 119–126.
Buckley, M. G., McEuen, A. R., and Walls, A. F. (1999) The detection of mast cell subpopulations in formalin-fixed human tissues using a new monoclonal antibody specific for chymase. J. Pathol. 189, 138–143.
Walls, A. F. (2000) The roles of neutral proteases in asthma and rhinitis. In Asthma and Rhinitis (Busse, W. W. and Holgate, S. T. eds.), 2nd Ed., Blackwell, Boston, pp. 968–998.
Beil, W. J., Schulz, M., McEuen, A. R., Buckley, M. G., and Walls, A. F. (1997) Number, fixation properties, dye-binding and protease expression of duodenal mast cells: comparisons between healthy subjects and patients with gastritis or Crohn’s disease. Histochem. J. 29, 759–773.
Li, L., Reddel, S. W., Cherrian, M., et al. (1998) Identification of basophilic cells that express mast cell granule proteases in the peripheral blood of asthma, allergy and drug-reactive patients. J. Immunol. 161, 5079–5086.
McEuen, A. R., Buckley, M. G., Compton, S. J., and Walls, A. F. (1999) Development and characterization of a monoclonal antibody specific for human basophils and the identification of a unique secretory product of basophil activation. Lab. Investig. 79, 27–38.
Claman, H. N., Choi, K. L., Sujansky, W., and Vatter, A. E. (1986) Mast cell “disappearance” in chronic murine graft-vs-host disease (GVHD)-ultrastructural demonstration of “phantom mast cells”. J. Immunol. 137, 2009–2013.
Mayrhofer, G., Gadd, S. J., Spargo, L. D., and Ashman, L. K. (1987) Specificity of a mouse monoclonal antibody raised against acute myeloid leukaemia cells for mast cells in human mucosal and connective tissues. Immunol. Cell Biol. 65, 241–250.
Schwartz, L. B., Irani, A.-M. A., Roller, K., Castells, M. C., and Schechter, N. M. (1987) Quantitation of histamine, tryptase and chymase in dispersed human T and TC mast cells. J. Immunol. 138, 2611–2615.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2008 Humana Press Inc., Totowa, NJ
About this protocol
Cite this protocol
Buckley, M., Walls, A.F. (2008). Identification of Mast Cells and Mast Cell Subpopulations. In: Jones, M.G., Lympany, P. (eds) Allergy Methods and Protocols. Methods in Molecular Medicine, vol 138. Humana Press. https://doi.org/10.1007/978-1-59745-366-0_24
Download citation
DOI: https://doi.org/10.1007/978-1-59745-366-0_24
Publisher Name: Humana Press
Print ISBN: 978-0-89603-896-7
Online ISBN: 978-1-59745-366-0
eBook Packages: Springer Protocols