Transforming growth factor-beta 1 (TGF-beta 1) promotes IL-2 mRNA expression through the up-regulation of NF-kappa B, AP-I and NF-AT in EL4 cells

Abstract

Transforming growth factor beta 1 (TGF-beta 1) has been previously shown to modulate interleukin 2 (IL-2) secretion by activated T-cells. In the present studies, we determined that TGF-beta 1 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-beta 1 (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-beta 1 on the activity of nuclear factor AT (NF-AT), nuclear factor kappa B (NF-kappa B), 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-beta 1 markedly increased NF-AT, NF-kappa B 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-kappa B)(3)-CAT, p(NF-AT)(3)-CAT and p(AP-1)(3)-CAT, TGF-beta 1 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-beta 1, 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-kappa B and AP-1, to help promote IL-2 mRNA expression by EL4 cells.