The recombinant expression vectors are designed to express human engineered T cell receptor (TCR) activator and human PD-L2 (GenBank Accession #NM_025239) in mammalian cells. The transfected cells can be used in conjunction with PD-1/NFAT Reporter/Jurkat T cells (BPS #60535) to study the interactions of PD-1 with PD-L2 ligａnd in a cellular context and screen for modulators of this signaling pathway.

In this assay, PD-1/NFAT Reporter/Jurkat T cells are used as effector cells; HEK293 cells overexpressing PD-L1 (or PD-L2) and an engineered T cell receptor (TCR) activator are used as target cells. When these two cells are co-c*ted, TCR complexes on effector cells are activated by TCR activator on target cells, resulting in expression of the NFAT luciferase reporter. However, PD1 and PD-L1 or PD-L2 ligation prevents TCR activation and suppresses the NFAT-responsive luciferase activity. This inhibition can be specifically reversed by anti-PD1 or anti-PD-L1 antibodies. PD1, PD-L1, or PD-L2 neutralizing antibodies block PD1:PD-L1 and/or PD1:PD-L2 interaction and promote T cell activation, resulting in reactivation of the NFAT responsive luciferase reporter.

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.