Early growth response genes 2 and 3 induced by AP-1 and NF-κB modulate TGF-β1 transcription in NK1.1− CD4+ NKG2D+ T cells

NK1.1⁻ CD4⁺ NKG2D⁺ T cells are a subpopulation of regulatory T cells that downregulate the functions of CD4⁺ T, CD8⁺ T, natural killer (NK) cells, and macrophages through TGF-β1 production. Early growth response genes 2 (Egr2) and 3 (Egr3) maintain immune homeostasis by modulating T lymphocyte development, inhibiting effector T cell function, and promoting the induction of regulatory T cells. Whether Egr2 and Egr3 directly regulate TGF-β1 transcription in NK1.1⁻ CD4⁺ NKG2D⁺ T cells remains elusive. The expression levels of Egr2 and Egr3 were higher in NK1.1⁻ CD4⁺ NKG2D⁺ T cells than in NK1.1⁻ CD4⁺ NKG2D⁻ T cells. Egr2 and Egr3 expression were remarkably increased after stimulating NK1.1⁻ CD4⁺ NKG2D⁺ T cells with sRAE or α-CD3/sRAE. The ectopic expression of Egr2 or Egr3 resulted in the enhancement of TGF-β1 expression, while knockdown of Egr2 or Egr3 led to the decreased expression of TGF-β1 in NK1.1⁻ CD4⁺ NKG2D⁺ T cells. Egr2 and Egr3 directly bound with the TGF-β1 promoter as demonstrated by the electrophoretic mobility shift assay and dual-luciferase gene reporter assay. Furthermore, the Egr2 and Egr3 expression of NK1.1⁻ CD4⁺ NKG2D⁺ T cells could be induced by the AP-1 and NF-κB transcriptional factors, but had no involvement with the activation of NF-AT and STAT3. In conclusion, Egr2 and Egr3 induced by AP-1 and NF-κB directly initiate TGF-β1 transcription in NK1.1⁻ CD4⁺ NKG2D⁺ T cells. This study indicates that manipulating Egr2 and Egr3 expression would potentiate or alleviate the regulatory function of NK1.1⁻ CD4⁺ NKG2D⁺ T cells and this strategy could be used in the therapy for patients with autoimmune diseases or tumor.