Mast Cells pp 363-381 | Cite as

In Vivo Models for Studying Mast Cell-Dependent Responses to Bacterial Infection

  • Christopher P. Shelburne
  • James B. McLachlan
  • Soman N. Abraham
Part of the Methods in Molecular Biology book series (MIMB, volume 315)


Mast cells are a critical component of host defense against bacterial infections. Activation of these cells during infection induces both innate and adaptive aspects of protective immunity needed for the elimination of the bacteria and survival of the host. These functional roles for the mast cell have been principally characterized using two in vivo models of acute bacterial infection featuring Gram-negative pathogens such as Escherichia coli. Here, we present basic protocols for the identification of mast cell-dependent biological functions during bacterial infection. These include the use of mast cell-deficient mice, the identification of mast cells in tissue, the culture of uropathogenic E. coli, and the basic analysis of mast cell-dependent functions in the peritoneal cavity and footpad models of bacterial pathogenesis.

Key Words

Mast cell bacteria E. coli UPEC tumor necrosis factor peritoneal cavity footpad draining popliteal lymph node 



This work was supported by funds from the National Institutes of Health DK50814 and AI50021 and the Sandler Foundation for Asthma Research.


  1. 1.
    Galli, S. J. and Hammel, I. (1994) Mast cell and basophil development. Curr. Optin. Hematol. 1, 33–39.Google Scholar
  2. 2.
    Wershil, B. K., Furuta, G. T., Wang, Z. S., and Galli, S. J. (1996) Mast cell-dependent neutrophil and mononuclear cell recruitment in immunoglobulin E-induced gastric reactions in mice. Gastroenterology 110, 1482–1490.CrossRefPubMedGoogle Scholar
  3. 3.
    Wershil, B. K., Wang, Z. S., Gordon, J. R., and Galli, S. J. (1991) Recruitment of neutrophils during IgE-dependent cutaneous late phase reactions in the mouse is mast cell-dependent. Partial inhibition of the reaction with antiserum against tumor necrosis factor-alpha. J. Clin. Invest. 87, 446–453.CrossRefPubMedGoogle Scholar
  4. 4.
    Secor, V. H., Secor, W. E., Gutekunst, C. A., and Brown, M. A. (2000) Mast cells are essential for early onset and severe disease in a murine model of multiple sclerosis. J. Exp. Med. 191, 813–822.CrossRefPubMedGoogle Scholar
  5. 5.
    Lee, D. M., Friend, D. S., Gurish, M. F., Benoist, C., Mathis, D., and Brenner, M. B. (2002) Mast cells: a cellular link between autoantibodies and inflammatory arthritis. Science 297, 1689–1692.CrossRefPubMedGoogle Scholar
  6. 6.
    Chen, R., Fairley, J. A., Zhao, M. L., et al. (2002) Macrophages, but not T and B lymphocytes, are critical for subepidermal blister formation in experimental bullous pemphigoid: macrophage-mediated neutrophil infiltration depends on mast cell activation. J. Immunol. 169, 3987–3992.PubMedGoogle Scholar
  7. 7.
    Marshall, J. S. and Bienenstock, J. (1994) The role of mast cells in inflammatory reactions of the airways, skin and intestine. Curr. Opin. Immunol. 6, 853–859.CrossRefPubMedGoogle Scholar
  8. 8.
    Malaviya, R., Ikeda, T., Ross, E., and Abraham, S. N. (1996) Mast cell modulation of neutrophil influx and bacterial clearance at sites of infection through TNFalpha. Nature 381, 77–80.CrossRefPubMedGoogle Scholar
  9. 9.
    Echtenacher, B., Mannel, D. N., and Hultner, L. (1996) Critical protective role of mast cells in a model of acute septic peritonitis. Nature 381, 75–77.CrossRefPubMedGoogle Scholar
  10. 10.
    McLachlan, J. B., Hart, J. P., Pizzo, S. V., et al. (2003) Mast cell-derived tumor necrosis factor induces hypertrophy of draining lymph nodes during infection. Nat. Immunol. 4, 1199–1205.CrossRefPubMedGoogle Scholar
  11. 11.
    Galli, S. J., Tsai, M., and Wershil, B. K. (1993) The c-kit receptor, stem cell factor, and mast cells. What each is teaching us about the others. Am. J. Pathol. 142, 965–974.PubMedGoogle Scholar
  12. 12.
    Nakano, T., Sonoda, T., Hayashi, C., et al. (1985) Fate of bone marrow-derived cultured mast cells after intracutaneous, intraperitoneal, and intravenous transfer into genetically mast cell-deficient W/Wv mice. Evidence that cultured mast cells can give rise to both connective tissue type and mucosal mast cells. J. Exp. Med. 162, 1025–1043.CrossRefPubMedGoogle Scholar
  13. 13.
    Ihle, J. N., Keller, J., Oroszlan, S., et al. (1983) Biologic properties of homogeneous interleukin 3. I. Demonstration of WEHI-3 growth factor activity, mast cell growth factor activity, p cell-stimulating factor activity, colony-stimulating factor activity, and histamine-producing cell-stimulating factor activity. J. Immunol. 131, 282–287.PubMedGoogle Scholar
  14. 14.
    Ryan, J. J., DeSimone, S., Klisch, G., et al. (1998) IL-4 inhibits mouse mast cell Fc epsilonRI expression through a STAT6-dependent mechanism. J. Immunol. 161, 6915–6923.PubMedGoogle Scholar
  15. 15.
    Galli, S. J., Wedemeyer, J., and Tsai, M. (2002) Analyzing the roles of mast cells and basophils in host defense and other biological responses. Int. J. Hematol. 75, 363–369.CrossRefPubMedGoogle Scholar
  16. 16.
    Weidner, N. and Austen, K. F. (1993) Heterogeneity of mast cells at multiple body sites. Fluorescent determination of avidin binding and immunofluorescent determination of chymase, tryptase, and carboxypeptidase content. Pathol. Res. Pract. 189, 156–162.PubMedGoogle Scholar
  17. 17.
    Biedermann, T., Kneilling, M., Mailhammer, R., et al. (2000) Mast cells control neutrophil recruitment during T cell-mediated delayed-type hypersensitivity reactions through tumor necrosis factor and macrophage inflammatory protein 2. J. Exp. Med. 192, 1441–1452.CrossRefPubMedGoogle Scholar
  18. 18.
    Anderson, W. H., Davidson, T. M., and Broide, D. H. (1996) Mast cell TNF mRNA expression in nasal mucosa demonstrated by in situ hybridization: a comparison of mast cell detection methods. J. Immunol. Methods 189, 145–155.CrossRefPubMedGoogle Scholar
  19. 19.
    Prodeus, A. P., Zhou, X., Maurer, M., Galli, S. J., and Carroll, M. C. (1997) Impaired mast cell-dependent natural immunity in complement C3-deficient mice. Nature 390, 172–175.CrossRefPubMedGoogle Scholar
  20. 20.
    Gommerman, J. L., Oh, D. Y., Zhou, X., et al. (2000) A role for CD21/CD35 and CD19 in responses to acute septic peritonitis: a potential mechanism for mast cell activation. J. Immunol. 165, 6915–6921.PubMedGoogle Scholar
  21. 21.
    Gordon, J. R. and Galli, S. J. (1991) Release of both preformed and newly synthesized tumor necrosis factor alpha (TNF-alpha)/cachectin by mouse mast cells stimulated via the Fc epsilon RI. A mechanism for the sustained action of mast cell-derived TNF-alpha during IgE-dependent biological responses. J. Exp. Med. 174, 103–107.CrossRefPubMedGoogle Scholar
  22. 22.
    Itano, A. A., McSorley, S. J., Reinhardt, R. L., et al. (2003) Distinct dendritic cell populations sequentially present antigen to CD4 T cells and stimulate different aspects of cell-mediated immunity. Immunity 19, 47–57.CrossRefPubMedGoogle Scholar
  23. 23.
    Shelburne, C. P., McCoy, M. E., Piekorz, R., et al. (2003) Stat5 expression is critical for mast cell development and survival. Blood 102, 1290–1297.CrossRefPubMedGoogle Scholar
  24. 24.
    Marshall, J. S., King, C. A., and McCurdy, J. D. (2003) Mast cell cytokine and chemokine responses to bacterial and viral infection. Curr. Pharm. Des. 9, 11–24.CrossRefPubMedGoogle Scholar

Copyright information

© Humana Press Inc. 2006

Authors and Affiliations

  • Christopher P. Shelburne
    • 1
  • James B. McLachlan
    • 1
  • Soman N. Abraham
    • 2
  1. 1.Department of PathologyDuke UniversityDurham
  2. 2.Department of Pathology, Molecular Genetics and Microbiology and ImmunologyDuke University Medical CenterDurham

Personalised recommendations