PD-1 / NFAT Reporter - Jurkat Recombinant Cell Line
PD-1/NFAT Reporter Jurkat Recombinant Cell Line is a Jurkat cell line that expresses human PD-1 (Programmed Cell Death 1, also known as PDCD1, SLEB2, CD279, GenBank Accession #NM_005018), and the firefly luciferase reporter 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.
PD-1 expression was verified by flow cytometry and this cell line was functionally validated with Anti-PD-1 and Anti-PD-L1 neutralizing antibodies in co-culture assays.
Figure 1: Illustration of the mechanism of action of PD-1/NFAT Reporter Jurkat Recombinant Cell Line in a co-culture assay.
The TCR (T cell receptor) activator present at the surface of PD-L1/TCR Activator CHO cells stimulate the TCRs in Jurkat T cells, whereas overexpression of PD-L1 on the CHO cell line engages Jurkat PD-1, blocking TCR activation signaling and preventing 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 complex and results in TCR activation and increased NFAT activity, which translates into increased luciferase reporter signal.
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Media Required for Cell Culture
Name | Ordering Information |
Thaw Medium 2 | BPS Bioscience #60184 |
Growth Medium 2A | BPS Bioscience #60190 |
Materials Required for Cellular Assay
Name | Ordering Information |
PD-L1 / TCR Activator - CHO Recombinant Cell Line | BPS Bioscience #60536 |
NFAT Luciferase Reporter Jurkat Cell Line | BPS Bioscience #60621 |
Thaw Medium 3 | BPS Bioscience #60186 |
Anti-PD-1 Neutralizing Antibody | BPS Bioscience #71120 |
Nivolumab (anti-PD-1) | SelleckChem #A2002 |
Anti-PD-L1 Neutralizing Antibody | BPS Bioscience #71213 |
ONE-Step™ Luciferase Assay System | BPS Bioscience #60690 |
96-well tissue culture-treated white clear-bottom assay plate | |
Luminometer |
The cell line has been screened to confirm the absence of Mycoplasma species.
PD-L1 and PD-L2 binding to PD-1, a receptor expressed on T-cells, negatively regulates immune responses. The PD-1 ligands PD-L1 and PD-L2 are found on the surface of most cancer cells, and their interaction with the receptor PD-1 inhibits T cell activity and allows cancer cells to escape immune surveillance. This pathway is also involved in regulating autoimmune responses. Therefore, these proteins (termed immune checkpoints) are promising therapeutic targets for many types of cancer as well as multiple sclerosis, arthritis, lupus, and type I diabetes. Checkpoint inhibitors have remarkable efficacy in a wide range of cancer types and have revolutionized cancer treatment. The PD-1 inhibitors nivolumab, pembrolizumab, cemiplimab and PD-L1 inhibitors atezolizumab, avelumab, and durvalumab are all FDA-approved drugs for immuno-therapy.