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TIGIT, PVRIG, and CD96 As Potential Immune Checkpoint Targets

The TIGIT, DNAM-1, CD96, and PVRIG Signaling Network

TIGIT, DNAM-1/CD226, CD96, and PVRIG are immunoglobulin superfamily (IgSF) receptors that share CD155/PVR and/or CD112/Nectin-2 as ligands, but mediate opposing effects on lymphocyte functions. While DNAM-1/CD226 acts as a co stimulatory receptor, TIGIT, PVRIG, and CD96 have all been suggested to function as co-inhibitory receptors. Further details about each of these receptors, their immuno-regulatory effects, and their potential as next generation immuno-oncology targets are described in the sections below.

Bio-Techne offers R&D SystemsTM bioactive recombinant proteins and antibodies for investigating the interactions between TIGIT, DNAM-1/CD226, CD96, PVRIG and their ligands, along with the immunoregulatory effects associated with these receptors. Our portfolio includes Fc, His, and Avi-tag biotinylated recombinant proteins for a variety of species and both unlabeled and fluorochrome-conjugated antibodies that are validated for multiple applications, including blocking/neutralization, flow cytometry, CyTOF, immunocytochemistry/immunofluorescence, and Western blot. 

 

Ligand-Receptor Interactions of the TIGIT, DNAM-1, CD96, PVRIG Signaling Network

TIGIT, PVRIG, and CD96 bind to CD112 and/or CD155 and inhibit T cell and natural killer cell functions.

The TIGIT, PVRIG, CD96, and DNAM-1/CD226 proteins bind to CD155/PVR and/or CD112 and have opposing effects on lymphocyte functions. TIGIT, PVRIG, CD96, and DNAM-1/CD226 are immunoglobulin superfamily receptors that mediate opposing effects on lymphocyte functions following ligand binding. All of these receptors bind to one or more of the following nectin or nectin-like molecules, CD111/Nectin-1, CD112/Nectin-2, CD113/Nectin-3, and CD155/PVR, which are frequently overexpressed on a broad range of tumors. The TIGIT protein binds to CD155/PVR, CD112 and CD113 and inhibits both T cell and NK cell activity. Conversely, DNAM-1/CD226 binds to CD155/PVR and CD112 and promotes CD8+ T cell and NK cell activity. PVRIG and CD96/TACTILE are additional T cell and NK cell inhibitory receptors that compete with DNAM-1/CD226 for binding to CD112 or CD155/PVR, respectively. Although the interaction between CD155/PVR and CD96/TACTILE has been shown to inhibit CD8+ T cell and NK cell functions in mice, whether this interaction mediates co-inhibitory or co-stimulatory effects in humans is still being investigated. In addition to its expression on T cells and NK cells, TIGIT may also be expressed on tumor cells and interact with T cell- or NK cell-expressed CD155/PVR, leading to a down-regulation of T cell and NK cell activity.

Analysis of the Binding Properties of R&D Systems Human TIGIT:CD155 Proteins by SPR

Surface plasmon resonance data showing the affinity measurements and binding kinetics between the CD155 and TIGIT proteins.

Affinity Measurements and Binding Kinetics of the CD155/PVR:TIGIT Protein Interaction by Surface Plasmon Resonance. Sensorgram data of captured Avi-tag Biotinylated Recombinant Human CD155/PVR Fc Chimera (R&D Systems, Catalog # AVI9174) binding to Recombinant Human TIGIT His-tag (R&D Systems, Catalog # 9525-TG). The corresponding overlaid kinetic fits with the residual plot shown below. The concentration of Recombinant Human TIGIT His-tag ranged from 0.2 nM to 400 nM. The corresponding steady state affinity fit is shown at the bottom. The experiment was performed on a Biacore T200, GE Healthcare.

Analysis of the Binding of R&D Systems Recombinant Human CD155 and DNAM-1 Proteins

Analysis of the binding response between R&D Systems Recombinant Human DNAM-1  and Recombinant Human CD155 proteins.

Recombinant Human CD155/PVR Protein Binds to Recombinant Human DNAM-1/CD226 Protein. Avi-tag Biotinylated Recombinant Human DNAM-1/CD226(R&D Systems, Catalog # AVI9298) was immobilized at 250 ng/mL on a Streptavidin-coated plate (R&D Systems, Catalog # CP004) and the indicated concentrations of Recombinant Human CD155/PVR Fc Chimera (R&D Systems, Catalog # 9174-CD) were added. Recombinant Human CD155/PVR Fc Chimera bound with an ED50 of 20-100 ng/mL.

Analysis of the Binding of R&D Systems Recombinant Human CD155 and CD96 Proteins

Analysis of the binding response between R&D Systems Recombinant Human CD96 and Recombinant Human CD155 proteins.

Recombinant Human CD155/PVR Protein Binds to Recombinant Human CD96 Protein. Recombinant Human CD155/PVR Fc Chimera (R&D Systems, Catalog # 9174-CD) was immobilized at 1 ug/mL and the indicated concentrations of Recombinant Human CD96 Fc Chimera (R&D Systems, Catalog # 9360-CD) were added. Recombinant Human CD96 Fc Chimera bound with an ED50 of 0.3-1.8 ug/mL.

Analysis of the Binding of R&D Systems Recombinant Human CD112 and PVRIG Proteins

Analysis of the binding response between R&D Systems Recombinant Human PVRIG and Recombinant Human CD112 proteins.

Recombinant Human Nectin-2/CD112 Protein Binds to Recombinant Human PVRIG Protein. Recombinant Human PVRIG Fc Chimera (R&D Systems, Catalog # 9365-PV) was immobilized at 1 ug/mL and the indicated concentrations of Recombinant Human Nectin-2/CD112 (R&D Systems, Catalog # 2229-N2) were added. Recombinant Human Nectin-2/CD112 bound with an ED50 of 0.4-2.4 ug/mL.

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