Unexpected characteristics of the IFN-γ reporters in nontransformed T cells

H. Zhu, J. Yang, T. L. Murphy, W. Ouyang, F. Wagner, A. Saparov, C. T. Weaver, K. M. Murphy

Research output: Contribution to journalArticle

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Abstract

Analysis of the IFN-γ promoter has primarily been conducted by transient expression of reporter constructs in transformed cells. However, the activity of cis elements may differ when expressed transiently compared with their activity within native chromatin. Furthermore, the transcription factors and signaling mechanisms in transformed cells may differ from those in normal T cells. To analyze IFN-γ promoter regulation in normal T cells, we developed a novel retroviral bottom-strand reporter system to allow the chromatin integration of promoter regions in primary developing T cells. As controls, both the IL-2 and IL-4 promoters were inducible in this system, with the IL-4 reporter having Th2-specific activity. Strikingly, the IFN-γ promoter exhibited constitutive activity in both Th1 and Th2 subsets, in contrast to the behavior of the endogenous IFN-γ gene, which is inducible only in Th1 cells. In mapping this activity, we found that the AP-1/GM-CSF site in the distal promoter element is the most critical element for the constitutive activity. Transgenic reporter lines for the IFN-γ promoter confirmed the constitutive behavior of the isolated IFN-γ promoter. This constitutive activity was resistant to inhibition by cyclosporin A and was independent of Stat4 and p38 mitogen-activated protein kinase. These results suggest that IFN-γ promoter regulation may require cis elements residing either downstream or >3.4 kb upstream of the transcriptional start site, involving repression of constitutive activity.

LanguageEnglish
Pages855-865
Number of pages11
JournalJournal of Immunology
Volume167
Issue number2
StatePublished - Jul 15 2001
Externally publishedYes

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T-Lymphocytes
Interleukin-4
Chromatin
Th1 Cells
Transcription Factor AP-1
p38 Mitogen-Activated Protein Kinases
Granulocyte-Macrophage Colony-Stimulating Factor
Genetic Promoter Regions
Cyclosporine
Interleukin-2
Transcription Factors
Genes

ASJC Scopus subject areas

  • Immunology

Cite this

Zhu, H., Yang, J., Murphy, T. L., Ouyang, W., Wagner, F., Saparov, A., ... Murphy, K. M. (2001). Unexpected characteristics of the IFN-γ reporters in nontransformed T cells.

Unexpected characteristics of the IFN-γ reporters in nontransformed T cells. / Zhu, H.; Yang, J.; Murphy, T. L.; Ouyang, W.; Wagner, F.; Saparov, A.; Weaver, C. T.; Murphy, K. M.

In: Journal of Immunology, Vol. 167, No. 2, 15.07.2001, p. 855-865.

Research output: Contribution to journalArticle

Zhu, H, Yang, J, Murphy, TL, Ouyang, W, Wagner, F, Saparov, A, Weaver, CT & Murphy, KM 2001, 'Unexpected characteristics of the IFN-γ reporters in nontransformed T cells' Journal of Immunology, vol. 167, no. 2, pp. 855-865.
Zhu H, Yang J, Murphy TL, Ouyang W, Wagner F, Saparov A et al. Unexpected characteristics of the IFN-γ reporters in nontransformed T cells. Journal of Immunology. 2001 Jul 15;167(2):855-865.
Zhu, H. ; Yang, J. ; Murphy, T. L. ; Ouyang, W. ; Wagner, F. ; Saparov, A. ; Weaver, C. T. ; Murphy, K. M./ Unexpected characteristics of the IFN-γ reporters in nontransformed T cells. In: Journal of Immunology. 2001 ; Vol. 167, No. 2. pp. 855-865
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