PD-L2/TCR Activator Mammalian Expression Kit
The TCR Activator/PD-L2 Mammalian Expression Kit is designed for the creation of engineered cells expressing TCR Activator and PD-L2. The kit consists of transfection-ready vectors encoding TCR activator and human PD-L2 that can be used to transfect cells and create target cells that overexpress PD-L2 and an engineered cell surface T cell receptor (TCR) activator. The kit contains enough plasmids for 500 assays if using a 96-well plate. TCR Activator expression vector alone is also provided as control for cellular experiments involving PD-L2.
Figure 1: Illustration of the principle of a PD-1:PD-L1/PD-L2 cellular assay.
Jurkat T cells expressing NFAT reporter with constitutive expression of PD-1 (PD-1/NFAT Reporter Jurkat Cell Line, BPS Bioscience #60535) act as effector cells. When co-cultivated, TCR complexes on PD-1/NFAT Reporter Jurkat cells are activated by the TCR activator expressed in the transfected cell line of interest, resulting in expression of the NFAT luciferase reporter. However, PD-1 and PD-L1/PD-L2 ligation prevents TCR activation and suppresses the NFAT-responsive luciferase activity. This inhibition can be specifically reversed by anti- PD-1 or anti- PD-L1/PD-L2 antibodies. Neutralizing antibodies block PD-1:PD-L1/PD-L2 interaction and result in reactivation of the NFAT responsive luciferase reporter.
Note: For assays in more difficult to transfect cell types, we suggest using our lentiviral products. The corresponding lentiviral product for this kit is BPS Bioscience #79894.
Name | Ordering Information |
Cell line of interest, such as HEK293 cells, and corresponding culture media | |
Transfection Reagent, such as Lipofectamine™ 2000 Transfection Reagent | Thermo Fisher #11668027 |
Opti-MEM I reduced Serum Medium | Thermo Fisher #31985-062 |
PD-1/NFAT Reporter Jurkat Cell Line | BPS Bioscience #60535 |
Anti-PD-1 Neutralizing Antibody | BPS Bioscience #71120 |
ONE-Step™ Luciferase Assay System | BPS Bioscience #60690 |
White clear-bottom 96-well cell culture plate | |
Luminometer |
Assay Medium:
RPMI1640 + 10% FBS + 1% Penicillin/Streptomycin.
Catalog # | Name | Amount | Storage |
TCR Activator + Human PD-L2 (expression vectors for expressing TCR Activator and human PD-L1) (100 ng/µl) | 500 µl | -20°C | |
TCR Activator (expression vectors for expressing TCR Activator) (100 ng/µl) | 500 µl | -20°C |
PD-L1 and PD-L2 binding to PD-1, a receptor expressed on T-cells, negatively regulates immune responses. PD-1 ligands PD-L1 and PD-L2 are found on the surface of many cancer cells, and their interaction with 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. PD-1 inhibitors nivolumab, pembrolizumab, cemiplimab and PD-L1 inhibitors atezolizumab, avelumab, and durvalumab are all FDA-approved drugs for immuno-therapy.
Sasca D, et al. (2019) Blood 133: 2305-2319.