We were the first to demonstrate that stimulation of IL-3 dependent murine mast cell lines with calcium ionophores, or by crosslinking high affinity IgE receptors (FcepsilonRI) with antigen or anti- IgE, induces the production of IL-3, IL-4 and other cytokines. We have also found that crosslinking of FcepsilonRI on two freshly isolated in vivo cell populations in non-immune mice, resting peritoneal mast cells and putative mast cell precursors in bone marrow, also induces cytokine secretion. This newly recognized ability of mast cells to secrete cytokines suggests a much wider role in inflammation than previously anticipated. This grant intends to extend our focus, to assess cytokine production in normal mast cells and determine how cytokine production relates to the state of mast cell differentiation both ontogenetically and phenotypically. First, we will test for qualitative differences in cytokine production induced by FcepsilonRI crosslinking of mast cell populations at distinct stages of differentiation, including mast cell precursors, mast cells differentiating during short vs. long term culture of bone marrow or spleen with IL-3, mature peritoneal mast cells, and mast cell lines differentiating into connective tissue phenotype. Second, we will test for quantitative and qualitative differences in the capacity of agents to induce cytokine production and/or regulate FcepsilonRI-mediated cytokine production by mast cells. We will explore the nature of FcepsilonRI crosslinking required and will also determine whether complement anaphylatoxins, and tachykinins, such as substance P, activate cytokine production. Since mast cells may express class II MHC molecules, we will determine whether mast cells activate T cells and whether T cells directly activate cytokine secretion by mast cells. We will extend our preliminary studies indicating that IL-3 markedly enhances the capacity of FcepsilonRI crosslinking to induce cytokine production, and test the regulatory effects of combinations of IL-3 with IL-4 and TGF-beta. Finally, we will evaluate the capacity of pharmacologic agents (which inhibit mast cell histamine secretion) to regulate cytokine secretion. We will determine whether any of these agents differ from FCepsilonRI crosslinking in their capacity to induce a specific spectrum of cytokines, as FcepsilonRI crosslinking induces the production of IL-3, IL-4, IL-5, IL-6 but not IL-2 and interferon-gamma. These experiments should provide a fuller understanding of the potential major role of mast cell precursors in immunologically mediated inflammation.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
5R01AI027906-02
Application #
3142207
Study Section
Immunological Sciences Study Section (IMS)
Project Start
1990-07-01
Project End
1993-06-30
Budget Start
1991-07-01
Budget End
1992-06-30
Support Year
2
Fiscal Year
1991
Total Cost
Indirect Cost
Name
Johns Hopkins University
Department
Type
Schools of Medicine
DUNS #
045911138
City
Baltimore
State
MD
Country
United States
Zip Code
21218
Schroeder, J T; MacGlashan Jr, D W; Lichtenstein, L M (2001) Human basophils: mediator release and cytokine production. Adv Immunol 77:93-122
Schroeder, J T; Howard, B P; Jenkens, M K et al. (1998) IL-4 secretion and histamine release by human basophils are differentially regulated by protein kinase C activation. J Leukoc Biol 63:692-8
Redrup, A C; Howard, B P; MacGlashan Jr, D W et al. (1998) Differential regulation of IL-4 and IL-13 secretion by human basophils: their relationship to histamine release in mixed leukocyte cultures. J Immunol 160:1957-64
Schroeder, J T; MacGlashan Jr, D W; MacDonald, S M et al. (1997) Regulation of IgE-dependent IL-4 generation by human basophils treated with glucocorticoids. J Immunol 158:5448-54
Schroeder, J T; MacGlashan Jr, D W (1997) New concepts: the basophil. J Allergy Clin Immunol 99:429-33
Schroeder, J T; MacGlashan Jr, D W; Kagey-Sobotka, A et al. (1994) IgE-dependent IL-4 secretion by human basophils. The relationship between cytokine production and histamine release in mixed leukocyte cultures. J Immunol 153:1808-17
MacGlashan Jr, D; White, J M; Huang, S K et al. (1994) Secretion of IL-4 from human basophils. The relationship between IL-4 mRNA and protein in resting and stimulated basophils. J Immunol 152:3006-16
Keegan, A D; Pierce, J H; Artrip, J et al. (1991) Ligand stimulation of transfected and endogenous growth factor receptors enhances cytokine production by mast cells. EMBO J 10:3675-82
Seder, R A; Paul, W E; Dvorak, A M et al. (1991) Mouse splenic and bone marrow cell populations that express high-affinity Fc epsilon receptors and produce interleukin 4 are highly enriched in basophils. Proc Natl Acad Sci U S A 88:2835-9
Seder, R A; Plaut, M; Barbieri, S et al. (1991) Purified Fc epsilon R+ bone marrow and splenic non-B, non-T cells are highly enriched in the capacity to produce IL-4 in response to immobilized IgE, IgG2a, or ionomycin. J Immunol 147:903-9

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