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Immunology T cell Tools

Study the balance of Tregs and Th17 cells

Expression vectors and reporters

  • Overexpress Human/Mouse Foxp3
  • Overexpress Human/Mouse RORγt
  • Foxp3, RORγt and IL-17 reporters
  • Jurkat T cell lines

Monitor transcription network activity in T cells

The transcription signal transduction pathways in regulatory T cells and T helper 17 (Th17) cells determine the differentiation and abundance of these two critical T cell subtypes. For Tregs, the transcription activation of Foxp3 is central to proliferating Tregs and enabling immunologilcal tolerance. The inflammation pathway mediated by Th17 cells depends upon the transcription activation of the RORγt and the IL-17 genes. SBI has engineered lentivector promoter reporters for the mouse Foxp3, RORγt and IL-17 genes coupled to GFP and luciferase dual reporter formats. Human Jurkat T cells were transduced with lentivirus for the Foxp3, RORγt and IL-17 promoter reporters. Transcription activation was tested using stimulation through the addition of with PMA (5 ng/ml) and Ionomycin (500 ng/ml). Cells were harvested after 24 hours and luciferase activity measured for transcription activation responses. The schematics for the promoter reporter structures and transactivation data with PMA/Ionomycin are depicted below.

Promoter sequences and structure designs are based on the following publications:

Foxp3 promoter: Tone M, Greene MI, 2011, Zheng Y, et al., 2011 and Burgler S, et al., 2010.

RORγt promoter: Dang EV, et al., 2011, Lazarevic V, et al., 2011 and Ruan Q,et al., 2011.

IL-17 promoter: Zhang F, Meng G, Strober W, 2008.