Interleukin-4 (IL-4) is a potent cytokine with pleiotropic effects on humoral and cellular immunity, hematopoiesis, and inflammation. Mounting evidence suggests that IL-4 is a central cytokine in allergic diseases. The secretion and function of IL-4 have been studied in a variety of in vitro and in vivo systems, but little is known about the molecular factors regulating its synthesis. The human IL-4 gene has recently been cloned, but there is limited data to date regarding control of its expression. The overall goal of this proposal is to rigorously investigate the transcriptional regulation of the human interleukin-4 gene. We have designed oligonucleotide primers for isolation of the 5'-flanking region of the IL-4 promoter from a human genomic library. We will use a variety of complementary and powerful techniques including single base-pair mutagenesis, DNAse I footprinting, and nuclear protein binding assays to systematically delineate the cis-elements and trans-acting factors involved in both constitutive and inducible expression of the IL-4 gene. We will study expression of the promoter construct and regulatory nuclear proteins in a variety of cell types including murine and human basophils and mast cells. This proposal has the potential to lead to important new insights into the control of IL-4 synthesis while serving as an excellent vehicle to expand the principal investigator's knowledge of and training in immunology and molecular biology.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Physician Scientist Award (K11)
Project #
5K11AI001152-05
Application #
2671338
Study Section
Allergy & Clinical Immunology-1 (AITC)
Project Start
1994-08-01
Project End
1999-07-31
Budget Start
1998-08-01
Budget End
1999-07-31
Support Year
5
Fiscal Year
1998
Total Cost
Indirect Cost
Name
Johns Hopkins University
Department
Internal Medicine/Medicine
Type
Schools of Medicine
DUNS #
045911138
City
Baltimore
State
MD
Country
United States
Zip Code
21218
Cianferoni, A; Schroeder, J T; Kim, J et al. (2001) Selective inhibition of interleukin-4 gene expression in human T cells by aspirin. Blood 97:1742-9
Keen, J C; Sholl, L; Wills-Karp, M et al. (2001) Preferential activation of nuclear factor of activated T cells c correlates with mouse strain susceptibility to allergic responses and interleukin-4 gene expression. Am J Respir Cell Mol Biol 24:58-65
Casolaro, V; Keane-Myers, A M; Swendeman, S L et al. (2000) Identification and characterization of a critical CP2-binding element in the human interleukin-4 promoter. J Biol Chem 275:36605-11
Chen, R; Burke, T F; Cumberland, J E et al. (2000) Glucocorticoids inhibit calcium- and calcineurin-dependent activation of the human IL-4 promoter. J Immunol 164:825-32
Burke, T F; Casolaro, V; Georas, S N (2000) Characterization of P5, a novel NFAT/AP-1 site in the human IL-4 promoter. Biochem Biophys Res Commun 270:1016-23
Georas, S N; Cumberland, J E; Burke, T F et al. (1998) Stat6 inhibits human interleukin-4 promoter activity in T cells. Blood 92:4529-38