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Research ArticleArticle

Transforming Growth Factor-Beta 1 (TGF-β1) Promotes IL-2 mRNA Expression Through the Up-regulation of NF-κB, AP-1 and NF-AT in EL4 Cells

Seung H. Han, Sung Su Yea, Young J. Jeon, Kyu-H. Yang and Norbert E. Kaminski
Journal of Pharmacology and Experimental Therapeutics December 1998, 287 (3) 1105-1112;
Seung H. Han
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Sung Su Yea
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Young J. Jeon
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Kyu-H. Yang
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Norbert E. Kaminski
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Abstract

Transforming growth factor β1 (TGF-β1) has been previously shown to modulate interleukin 2 (IL-2) secretion by activated T-cells. In the present studies, we determined that TGF-β1 induced IL-2 mRNA expression in the murine T-cell line EL4, in the absence of other stimuli. IL-2 mRNA expression was significantly induced by TGF-β1 (0.1–1 ng/ml) over a relatively narrow concentration range, which led to the induction of IL-2 secretion. Under identical condition, we examined the effect of TGF-β1 on the activity of nuclear factor AT (NF-AT), nuclear factor κB (NF-κB), activator protein-1 (AP-1) and octamer, all of which contribute to the regulation of IL-2 gene expression. Electrophoretic mobility shift assays showed that TGF-β1 markedly increased NF-AT, NF-κB and AP-1 binding to their respective cognate DNA binding sites, whereas octamer binding remained constant, as compared with untreated cells. Employing a reporter gene expression system with p(NF-κB)3-CAT, p(NF-AT)3-CAT and p(AP-1)3-CAT, TGF-β1 treatment of transfected EL4 cells induced a dose-related increase in chloramphenicol acetyltransferase activity that correlated well with the DNA binding profile found in the electrophoretic mobility shift assay studies. These results show that TGF-β1, in the absence of any additional stimuli, up-regulates the activity of key transcription factors involved in IL-2 gene expression, including NF-AT, NF-κB and AP-1, to help promote IL-2 mRNA expression by EL4 cells.

Footnotes

  • Send reprint requests to: Dr. Norbert Kaminski, Department of Pharmacology & Toxicology, B440 Life Science Bldg., Michigan State University, East Lansing, MI 48824.

  • ↵1 This work was supported by NIEHS Superfund Grant PO1 P42ES04911-08C.

  • ↵2 Present address: Dept. of Biological Sciences, Korea Advanced Institute of Science and Technology, Taejon, Korea.

  • Abbreviations:
    NF-AT
    nuclear factor AT
    IL-2
    interleukin 2
    NF-κB
    nuclear factor κB
    AP-1
    activator protein-1
    Oct
    octamer
    FCS
    fetal calf serum
    EMSA
    electrophoretic mobility shift assay
    PMA
    phorbol-12-myristate-13-acetate
    CAT
    chloramphenicol acetyltransferase
    rcRNA
    recombinant RNA
    IS
    internal standard
    RT-PCR
    reverse transcriptase polymerase chain reaction
    FKBP12
    immunophilin-FKBP12
    TGF-β1
    transforming growth factor-beta 1
    lg
    immunoglobulin
    Th
    T helper cell
    IFN-γ
    interferon-gamma
    CD
    cluster designation
    RPMI
    Roswell Park Memorial Institute
    DEAE
    diethylaminoethyl
    • Received January 9, 1998.
    • Accepted July 1, 1998.
  • The American Society for Pharmacology and Experimental Therapeutics
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Journal of Pharmacology and Experimental Therapeutics
Vol. 287, Issue 3
1 Dec 1998
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Research ArticleArticle

Transforming Growth Factor-Beta 1 (TGF-β1) Promotes IL-2 mRNA Expression Through the Up-regulation of NF-κB, AP-1 and NF-AT in EL4 Cells

Seung H. Han, Sung Su Yea, Young J. Jeon, Kyu-H. Yang and Norbert E. Kaminski
Journal of Pharmacology and Experimental Therapeutics December 1, 1998, 287 (3) 1105-1112;

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Research ArticleArticle

Transforming Growth Factor-Beta 1 (TGF-β1) Promotes IL-2 mRNA Expression Through the Up-regulation of NF-κB, AP-1 and NF-AT in EL4 Cells

Seung H. Han, Sung Su Yea, Young J. Jeon, Kyu-H. Yang and Norbert E. Kaminski
Journal of Pharmacology and Experimental Therapeutics December 1, 1998, 287 (3) 1105-1112;
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