Abstract
Mast cells are granulated immune cells typically located at barrier sites of the body, such as the skin and the mucosa of the respiratory, urogenital, and gastrointestinal tract. They are well known for their capacity to participate in the orchestration of inflammatory and immune responses by releasing a broad array of mediators as a consequence of IgE-dependent and IgE-independent activation. Mast cells derive from myeloid progenitors, but in contrast to other myeloid cells, they leave the bone marrow in an immature state; therefore, mast cells are not visible in the blood under normal conditions. For full maturation, the tissue environment is necessary. Thus, mature mast cells can be only isolated from tissue such as skin or mucosal sites, which makes mast cell isolation complicated. This chapter describes methods to isolate, purify, and culture mast cells from the human intestinal mucosa. Human mucosal mast cells can be used to characterize their mediators and to study the mechanisms of human mast cell activation, signal transduction, and exocytosis in response to specific stimuli.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Similar content being viewed by others
References
Bischoff SC (2007) Role of mast cells in allergic and non-allergic immune responses: comparison of human and murine data. Nat Rev Immunol 7:93–104
Kalesnikoff J, Galli SJ (2008) New developments in mast cell biology. Nat Immunol 9:1215–1223
Galli SJ, Grimbaldeston M, Tsai M (2008) Immunomodulatory mast cells: negative, as well as positive, regulators of innate and acquired immunity. Nat Rev Immunol 8:478–486
Bischoff SC (2009) Physiological and pathophysiological functions of intestinal mast cells. Semin Immunopathol 31:185–205
Marshall JS (2004) Mast cell responses to pathogens. Nat Rev Immunol 4:787–799
Rivera J, Gilfillan AM (2006) Molecular regulation of mast cell activation. J Allergy Clin Immunol 117:1214–1225
Bischoff SC, Dahinden CA (1992) c-Kit ligand: a unique potentiator of mediator release by human lung mast cells. J Exp Med 175:237–244
Bischoff SC, Sellge G, Lorentz A, Sebald W, Raab R, Manns MP (1999) IL-4 enhances proliferation and mediator release in mature human mast cells. Proc Natl Acad Sci U S A 96:8080–8085
Lorentz A, Schwengberg S, Sellge G, Manns MP, Bischoff SC (2000) Human intestinal mast cells are capable of producing different cytokine profiles: role of IgE receptor cross-linking and IL-4. J Immunol 164:43–48
Babina M, Guhl S, Starke A, Kirchhof L, Zuberbier T, Henz BM (2004) Comparative cytokine profile of human skin mast cells from two compartments strong resemblance with monocytes at baseline but induction of IL-5 by IL-4 priming. J Leukoc Biol 75:244–252
Hundley TR, Gilfillan AM, Tkaczyk C, Andrade MV, Metcalfe DD, Beaven MA (2004) Kit and FcεRI mediate unique and convergent signals for release of inflammatory mediators from human mast cells. Blood 104:2410–2417
Lorentz A, Wilke M, Sellge G, Worthmann H, Klempnauer J, Manns MP, Bischoff SC (2005) IL-4 induced priming of human intestinal mast cells for enhanced survival and Th2 cytokine generation is reversible and associated with an increased activity of ERK1/2 and c-Fos. J Immunol 174:6751–6756
Feuser K, Feilhauer K, Staib L, Bischoff SC, Lorentz A (2011) Akt crosslinks IL-4 priming, stem cell factor signaling, and IgE dependent activation in mature human mast cells. Mol Immunol 48:546–552
Lorentz A, Schwengberg S, Mierke C, Manns MP, Bischoff SC (1999) Human intestinal mast cells produce IL-5 in vitro upon IgE receptor cross-linking and in vivo in the course of intestinal inflammatory disease. Eur J Immunol 29:1496–1503
He SH (2004) Key role of mast cells and their major secretory products in inflammatory bowel disease. World J Gastroenterol 10:309–318
Guhl S, Babina M, Neou A, Zuberbier T, Artuc M (2010) Mast cell lines HMC-1 and LAD2 in comparison with mature human skin mast cells—drastically reduced levels of tryptase and chymase in mast cell lines. Exp Dermatol 19:845–847
Kovarova M, Latour AM, Chason KD, Tilley SL, Koller BH (2010) Human embryonic stem cells: a source of mast cells for the study of allergic and inflammatory diseases. Blood 115:3695–3703
Rådinger M, Jensen BM, Kuehn HS, Kirshenbaum A, Gilfillan AM (2010) Generation, isolation, and maintenance of human mast cells and mast cell lines derived from peripheral blood or cord blood. Curr Protoc Immunol Chapter 7:Unit 7.37
Schulman ES, Macglashan DW, Peters SP, Schleimer RP, Newball HH, Lichtenstein LM (1982) Human lung mast cells: purification and characterization. J Immunol 129:2662–2667
Gibbs BF, Wierecky J, Welker P, Henz BM, Wolff HH, Grabbe J (2001) Human skin mast cells rapidly release preformed and newly generated TNF-alpha and IL-8 following stimulation with anti-IgE and other secretagogues. Exp Dermatol 10:312–320
Kulka M, Metcalfe DD (2010) Isolation of tissue mast cells. Curr Protoc Immunol Chapter 7:Unit 7.25
Befus AD, Dyck N, Goodacre R, Bienenstock J (1987) Mast-cells from the human intestinal lamina propria—isolation, histochemical subtypes, and functional-characterization. J Immunol 138:2604–2610
Lowman MA, Rees PH, Benyon RC, Church MK (1988) Human mast cell heterogeneity: histamine release from mast cells dispersed from skin, lung, adenoids, tonsils, and colon in response to IgE-dependent and nonimmunologic stimuli. J Allergy Clin Immunol 81:590–597
Bischoff SC, Schwengberg S, Raab R, Manns MP (1997) Functional properties of human intestinal mast cells cultured in a new culture system: enhancement of IgE receptor-dependent mediator release and response to stem cell factor. J Immunol 159:5560–5567
Bischoff SC, Sellge G, Schwengberg S, Lorentz A, Manns MP (1999) Stem cell factor-dependent survival, proliferation and enhanced releasability of purified mature mast cells isolated from human intestinal tissue. Int Arch Allergy Immunol 118:104–107
Gebhardt T, Sellge G, Lorentz A, Raab R, Manns MP, Bischoff SC (2002) Cultured human intestinal mast cells express functional IL-3 receptors and respond to IL-3 by enhancing growth and IgE receptor-dependent mediator release. Eur J Immunol 32:2308–2316
Sander LE, Frank SP, Bolat S, Blank U, Galli T, Bigalke H, Bischoff SC, Lorentz A (2008) Vesicle associated membrane protein (VAMP)-7 and VAMP-8, but not VAMP-2 or VAMP-3, are required for activation-induced degranulation of mature human mast cells. Eur J Immunol 38:855–863
Frank SP, Thon KP, Bischoff SC, Lorentz A (2011) SNAP-23 and syntaxin-3 are required for chemokine release by mature human mast cells. Mol Immunol 49(1–2):353–358
Walev I, Bhakdi SC, Hofmann F, Djonder N, Valeva A, Aktories K, Bhakdi S (2001) Delivery of proteins into living cells by reversible membrane permeabilization with streptolysin-O. Proc Natl Acad Sci U S A 98:3185–3190
Acknowledgments
The authors thank all former and current colleagues and in particular C. A. Dahinden, K. Wordelmann, S. Schwengberg, C. T. Mierke, G. Weier, T. Gebhardt, L. E. Sander, S. P. Frank, A. Mrasori, K. Feuser, and Y. Soltow who were involved in establishing the methods described here.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2015 Springer Science+Business Media New York
About this protocol
Cite this protocol
Lorentz, A., Sellge, G., Bischoff, S.C. (2015). Isolation and Characterization of Human Intestinal Mast Cells. In: Hughes, M., McNagny, K. (eds) Mast Cells. Methods in Molecular Biology, vol 1220. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-1568-2_11
Download citation
DOI: https://doi.org/10.1007/978-1-4939-1568-2_11
Published:
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-1567-5
Online ISBN: 978-1-4939-1568-2
eBook Packages: Springer Protocols