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PVRIG (CD112R) is a co-inhibitory receptor belonging to the Poliovirus Receptor (PVR) family, expressed on Natural Killer (NK) cells and T cells. It binds to its ligand (CD112, also known as PVRL2/nectin-2) to suppress the response of T cells and NK cells against cancer. Consequently, CD112R holds significant potential as a novel target for immune checkpoint inhibition in cancer immunotherapy. 

CD112 (Cluster of Differentiation 112)—also known as Nectin-2 or PVR-related protein 2 (PVRL2)—is a member of the Nectin family and is associated with tumor angiogenesis, growth, and metastasis. CD112 comprises an extracellular region containing three immunoglobulin (Ig)-like domains (two IgC domains and one distal IgV domain), a cytoplasmic tail, and a transmembrane region. CD112 is readily detectable on antigen-presenting cells or tumor cells; its high-level expression is correlated with tumor progression and poor prognosis in the majority of cancer patients. 

The binding of CD112R to its ligand recruits SHP1 and SHP2, thereby triggering the inhibition of TCR/CD28 signaling within T cells. PVRIG can also exert immunosuppressive effects by competitively binding to the ligand shared with CD226, thereby blocking the stimulatory signals transmitted by CD226. This principle of immunosuppression is analogous to that employed by TIGIT—another member of the same receptor family—and it is possible that the two receptors act synergistically to mediate immunosuppression.

The PVRIG (CD112R) Effector Reporter Cell model accurately simulates the in vivo signal transduction processes of PVRIG; the underlying principle is illustrated in the figure below.

Figure 1. Schematic Diagram of the PVRIG(CD112R) Effector Reporter Cell Model

Figure 2. Recombinant PVRIG(CD112R) Effector Reporter Cell stably expressing PVRIG(CD112R).

Figure 3. Dose Response of PVRIG Blocking Antibody in PVRIG(CD112R) Effector Reporter Cell (C7) with CD112 TCR Activator CHO (C1)

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.