Antagonist screening assay for PD-1/PD-L1 axis

InvivoGen offers a cellular assay specifically designed for screening antibody-, Fc-fusion protein-, or small-molecule antagonists of the PD-1/PD-L1 immune checkpoint (IC) axis. This assay is comprised of:

Principle of PD-1/PD-L1 cellular assay
(click to enlarge)

InvivoGen also offers:

• Raji-APC-Null Cells
• Anti-hPD-1 & hPD-L1 antibodies
• Anti-β-Gal antibodies

Jurkat-Lucia™ TCR-hPD-1: Reporter T cells
Raji-APC-hPD-L1: Antigen-presenting cells (APCs)

These paired cell lines allow the mimicking of the immune synapse between T cells and APCs through the interaction of cell surface molecules delivering T-cell activation and concomitant inhibitory checkpoint signals:
- Signal 1 is delivered upon recognition of a specific [HLA::peptide] complex on Raji APC cells by the T-cell receptor (TCR) on Jurkat effector cells,
- Signal 2 is operated by the interaction of CD80/86 and CD28 at the surface of the Raji APC and Jurkat effector cells, respectively,
- PD-1/PD-L1 IC signal is a regulatory signal that inhibits T-cell activation.

More details

Assay principle:

This assay relies on the co-culture of Jurkat-Lucia™ TCR-hPD-1 and Raji-APC-hPD-L1 cells.
Jurkat-Lucia™ TCR-hPD-1 cells express a specific TCR and the PD-1 inhibitory IC receptor along with the Lucia luciferase reporter gene under the control of an ISG54 minimal promoter fused to six NFAT response elements.
Raji-APC-hPD-L1 cells express a specific [HLA::peptide] complex and the PD-L1 inhibitory IC ligand.
In the presence of a potent PD-1 or PD-L1 antagonist, the IC-mediated inhibition is blocked and the TCR activation leads to Lucia production. Activation of the reporter T cells can be readily measured using QUANTI-Luc™ 4 Lucia/Gaussia detection reagent (see Figures).

T-cell key features:

Stable specific [HLA::peptide]-restricted TCR
Stable hCD28 overexpression
Stable hPD-1 overexpression
NFAT-inducible Lucia luciferase reporter activity

APC key features:

Stable specific [HLA::peptide] expression
Endogenous hCD80/86 expression
Stable hPD-L1 overexpression

Read our review on Immune Checkpoint Blockade

Learn more about Immune Checkpoint Antibodies.

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