The objective of this proposal is to characterize the soluble factors released by mononuclear cells that regulate the function of basophils and mast cells. These cells have a major role in immediate hypersensitivity reactions (asthma, rhinitis, anaphylaxis) caused by IgE antibodies. Basophils also accumulate locally in many other late onset reactions: contact dermatitis; rejection of tumors, microbes, transplants; and the chronic stage of IgE dependent reactions. It is postulated that mononuclear cells release cytokines that direct the migration of basophils and the secretion of basophils and mast cells. We have identified and characterized two human cytokines acting on basophils, a basophil chemotactic activity and a histamine-releasing activity. These factors will be prepared in large quantities by culture of leukocyte concentrates from normal blood donors and human T-cell lines. Factors will be purified by ion exchange, gel filtration, affinity and high performance liquid chromatographies. The cytokines will then be used for production of specific antibodies and for determining their effects on basophils and mast cells. Polyclonal antibodies will be prepared by immunizing rabbits with purified HRA in order to provide an assay for monoclonal antibodies to be prepared in mice. These antibodies will be used for affinity chromatography, characterization of cytokines, and measurements of HRA in biological fluids. The cytokines produced by unfractionated mononuclear cells will be compared with the products of purified T-cells, T-cell subsets and monocyte-macrophages. Comparisons to be made are physicochemical properties, antigenic cross-reactivity and functional response of basophils. One hypothesis to be tested is whether the same molecules can regulate both migration and secretion of basophils. We speculate that at low concentrations, cytokines induce cell movement, and at high concentrations, exocytosis. This will be tested by combining different concentrations of cytokines with solid or fluid phase antibodies, and then determining the function (chemotaxis and release of histamine) and morphologic (electron microscopy) responses of purified basophils. The biological significance of HRA will be studied by seeking this cytokine in blister fluid from patients with contact dermatitis, and in bronchoalveolar lavage fluid from asthmatics. The spontaneous and antigen-induced synthesis of HRA by cells from asthmatics cultured in vitro will be followed and levels correlated with the clinical course and response to immunotherapy.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
5R01AI022940-02
Application #
3134668
Study Section
Immunological Sciences Study Section (IMS)
Project Start
1986-09-01
Project End
1989-08-31
Budget Start
1987-09-01
Budget End
1988-08-31
Support Year
2
Fiscal Year
1987
Total Cost
Indirect Cost
Name
University of Texas Medical Br Galveston
Department
Type
Schools of Medicine
DUNS #
041367053
City
Galveston
State
TX
Country
United States
Zip Code
77555
Sim, T C; Reece, L M; Hilsmeier, K A et al. (1995) Secretion of chemokines and other cytokines in allergen-induced nasal responses: inhibition by topical steroid treatment. Am J Respir Crit Care Med 152:927-33
Chonmaitree, T; Patel, J A; Lett-Brown, M A et al. (1994) Virus and bacteria enhance histamine production in middle ear fluids of children with acute otitis media. J Infect Dis 169:1265-70
Sim, T C; Grant, J A; Hilsmeier, K A et al. (1994) Proinflammatory cytokines in nasal secretions of allergic subjects after antigen challenge. Am J Respir Crit Care Med 149:339-44
Sim, T C; Hilsmeier, K A; Reece, L M et al. (1994) Interleukin-1 receptor antagonist protein inhibits the synthesis of IgE and proinflammatory cytokines by allergen-stimulated mononuclear cells. Am J Respir Cell Mol Biol 11:473-9
Alam, R; Dejarnatt, A; Stafford, S et al. (1993) Selective inhibition of the cutaneous late but not immediate allergic response to antigens by misoprostol, a PGE analog. Results of a double-blind, placebo-controlled randomized study. Am Rev Respir Dis 148:1066-70
Sim, T C; Alam, R; Forsythe, P A et al. (1992) Measurement of histamine-releasing factor activity in individual nasal washings: relationship with atopy, basophil response, and membrane-bound IgE. J Allergy Clin Immunol 89:1157-65
Alam, R; Lett-Brown, M A; Forsythe, P A et al. (1992) Monocyte chemotactic and activating factor is a potent histamine-releasing factor for basophils. J Clin Invest 89:723-8
Alam, R; Forsythe, P A; Stafford, S et al. (1992) Macrophage inflammatory protein-1 alpha activates basophils and mast cells. J Exp Med 176:781-6
Sim, T C; Hilsmeier, K A; Alam, R et al. (1992) Effect of topical corticosteroids on the recovery of histamine releasing factors in nasal washings of patients with allergic rhinitis. A double-blind, randomized, placebo-controlled study. Am Rev Respir Dis 145:1316-20
Alam, R; Forsythe, P A; Lett-Brown, M A et al. (1992) Interleukin-8 and RANTES inhibit basophil histamine release induced with monocyte chemotactic and activating factor/monocyte chemoattractant peptide-1 and histamine releasing factor. Am J Respir Cell Mol Biol 7:427-33

Showing the most recent 10 out of 19 publications