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TIGIT—also known as WUCAM, Vstm3, or VSIG9—is a co-inhibitory receptor belonging to the immunoglobulin superfamily. High levels of TIGIT expression are observed on effector CD4+ and CD8+ T cells, regulatory T cells, and NK cells. TIGIT's ligands include CD112 and the Poliovirus Receptor (PVR—also known as CD155, Necl-5, and Tage4), with PVR serving as TIGIT's high-affinity cognate receptor. Furthermore, TIGIT competes with CD226 (DNAM-1) and CD96 (TACTILE) for ligands. TIGIT effectively blocks the binding of CD155 to either CD96 or CD226, further substantiating that TIGIT possesses the highest affinity for CD155. TIGIT not only competes with CD226 for ligands but can also directly bind to CD226 *in cis*, thereby preventing its homodimerization and rendering CD226 unable to bind to CD155 to exert its co-stimulatory function. 

PVRIG (CD112R) is a co-inhibitory receptor within the Poliovirus Receptor (PVR) family, expressed on Natural Killer (NK) cells and T cells. It binds to its ligand (CD112 or PVRL2/nectin-2) to suppress T-cell and NK-cell responses against cancer. Additionally, PVRIG exerts immunosuppressive effects by competitively binding to the ligand shared with CD226, thereby blocking the stimulatory signals transmitted by CD226. This mechanism of immunosuppression is similar to that of its family member, TIGIT; moreover, the two receptors may act synergistically to mediate immunosuppression. Consequently, CD112R holds significant potential as a novel immune checkpoint inhibitor in cancer immunotherapy. 

The TIGIT & PVRIG Dual Effector Reporter Cell model accurately simulates the *in vivo* signal transduction processes of TIGIT and PVRIG; the underlying principle is illustrated in the figure below.

Figure 1.TIGIT&PVRIG Dual Effector Reporter Cell Cell Model Schematic

Figure 2. Recombinant TIGIT/PVRIG Dual Effector Reporter Cell constitutively expressing TIGIT and PVRIG.

Figure 3. Dose Response of Blocking Antibodies in TIGIT/PVRIG Dual Effector Reporter Cell(C45) with CD112/CD155 Dual aAPC Cell.

Cell Passage Procedures

1.This cell line grows in suspension.
2.Upon receipt, cells should be thawed immediately or stored in liquid nitrogen until use.
3.Before thawing, pre-warm the water bath and culture medium to 37 °C, and prepare a small amount of dry ice.
4.Remove the cryovial from storage and transport it to the cell culture laboratory on dry ice.
5.Rapidly thaw the cells in a 37 °C water bath. Once the cells are completely thawed, spray the cryovial with 70% ethanol for disinfection and transfer it to a biosafety cabinet.
6.Add 10 mL of pre-warmed culture medium into a 15 mL centrifuge tube. Transfer the contents of the cryovial into the tube and centrifuge at 1000 rpm for 5 minutes.
7.Carefully discard the supernatant. Resuspend the cell pellet in 5 mL of pre-warmed culture medium by gentle pipetting. Immediately perform cell counting and adjust the cell density to 3–6 × 10⁵ cells/mL based on the counting results, then transfer the cells into a culture flask.
8.Count the cells every 1–2 days. When the cell density exceeds 1 × 10⁶ cells/mL, passage the cells promptly or add fresh culture medium. Maintain the cell density between 2 × 10⁵ and 1 × 10⁶ cells/mL.

Suspension Cell Cryopreservation Procedure:

1.Collect 8 × 10⁶ cells, centrifuge, and discard the supernatant.
2.Add 1 mL of cell freezing medium (90% FBS + 10% DMSO) and gently pipette to mix thoroughly. Transfer the suspension into a cryovial.
3.Immediately place the cryovial into a controlled-rate freezing container (Nalgene 5100-0001), fill with isopropanol up to the indicated level, and store at −80 °C.
4.After 24 hours, transfer the cryovial to liquid nitrogen for long-term storage.