Recombinant Jurkat T cells constitutively expressing human PD-1 (Programmed Cell Death 1, also known as PDCD1, SLEB2, CD279, GenBank Accession #NM_005018), and with conditional expression of the firefly luciferase gene under the control of NFAT response elements located upstream of the minimal TATA promoter. Stimulation of NFAT can therefore be monitored by measuring luciferase activity.

Illustration of a suggested co-culture experiment. The TCR activator present at the surface of TCRa/PD-L1 CHO cells stimulates TCR in Jurkat T cells, whereas overexpression of PD-L1 on the CHO cell line engages Jurkat PD-1, which blocks TCR activation signals and prevents activation of NFAT. Addition of a neutralizing anti-PD-1 or anti-PD-L1 antibody to the co-culture releases the PD-L1/PD-1 brake and results in TCR activation and increased NFAT activity, which translates into increased luciferase reporter signal.

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