Abstract
Mast cells are the crucial effector cells for allergic reactions. They are activated through the aggregation of the high-affinity immunoglobulin E (IgE) receptor (FcεRI) with allergen and allergen-specific IgE. Tyrosine phosphorylation of FcεRI subunits and various signaling proteins is an initial triggering event, leading to the activation of several signaling pathways in mast cells. Much has been learned from analysis of mast cells derived from gene-targeted mice. Therefore, in this chapter we will first describe how to generate mast cells from mouse bone marrow cells and how to correct the genetic defect by retroviral transduction. Then, we will describe how to assess early activation events by measuring several protein-tyrosine kinases and serine/threonine kinases such as protein kinase B (= Akt), protein kinase C, and JNK. As signal transduction is highly dependent on protein-protein interactions, we will describe experimental details of co-immunoprecipitation methods that are used to confirm such interactions.
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References
Galli, S. J. and Lantz, C. S. (1999) Allergy, in Fundamental Immunology (Paul, W. E., ed.), Lippincott-Raven Press, Philadelphia, PA, pp. 1137–1184.
Kinet, J. P. (1999) The high-affinity IgE receptor (Fc epsilon RI): from physiology to pathology. Annu. Rev. Immunol. 17, 931–972.
Turner, H. and Kinet, J. P. (1999) Signalling through the high-affinity IgE receptor Fc epsilonRI. Nature 402, B24–B30.
Kawakami, T. and Galli, S. J. (2002) Regulation of mast-cell and basophil function and survival by IgE. Nat. Rev. Immunol. 2, 773–786.
Kawakami, T., Inagaki, N., Takei, M., et al. (1992) Tyrosine phosphorylation is required for mast cell activation by Fc epsilon RI cross-linking. J. Immunol. 148, 3513–3519.
Hata, D., Kawakami, Y., Inagaki, N., et al. (1998) Involvement of Bruton’s tyrosine kinase in FcepsilonRI-dependent mast cell degranulation and cytokine production. J. Exp. Med. 187, 1235–1247.
Yokota, T., Lee, F., Arai, N., et al. (1986) Strategies for cloning mouse and human lymphokine genes using mammalian cDNA expression vector. Lymphokines 13, 1–19.
Onishi, M., Kinoshita, S., Morikawa, Y., et al. (1996) Applications of retrovirusmediated expression cloning. Exp. Hematol. 24, 324–329.
Kawakami, Y., Miura, T., Bissonnette, R., et al. (1997) Bruton’s tyrosine kinase regulates apoptosis and JNK/SAPK kinase activity. Proc. Natl. Acad. Sci. USA 94, 3938–3942.
Pear, W. S., Nolan, G. P., Scott, M. L., and Baltimore, D. (1993) Production of high-titer helper-free retroviruses by transient transfection. Proc. Natl. Acad. Sci. USA 90, 8392–8396.
Grignani, F., Kinsella, T., Mencarelli, A., et al. (1998) High-efficiency gene transfer and selection of human hematopoietic progenitor cells with a hybrid EBV/retroviral vector expressing the green fluorescence protein. Cancer Res. 58, 14–19.
Morita, S., Kojima, T., and Kitamura, T. (2000) Plat-E: an efficient and stable system for transient packaging of retroviruses. Gene Ther. 7, 1063–1066.
Yao, L., Kawakami, Y., and Kawakami, T. (1994) The pleckstrin homology domain of Bruton tyrosine kinase interacts with protein kinase C. Proc. Natl. Acad. Sci. USA 91, 9175–9179.
Wong, G. W., Friend, D. S., and Stevens, R. L. (1998) Mouse and rat models of mast cell development, in Signal Transduction in Mast Cells and Basophils, (Razin, E., and Rivera, J., eds.), Springer-Verlag, New York, NY, pp. 39–53.
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Kawakami, Y., Kitaura, J., Kawakami, T. (2006). Techniques to Study FcεRI Signaling. In: Krishnaswamy, G., Chi, D.S. (eds) Mast Cells. Methods in Molecular Biology, vol 315. Humana Press. https://doi.org/10.1385/1-59259-967-2:175
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DOI: https://doi.org/10.1385/1-59259-967-2:175
Publisher Name: Humana Press
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