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TIGIT—also known as WUCAM, Vstm3, or VSIG9—is a co-inhibitory receptor belonging to the immunoglobulin superfamily. TIGIT consists of an extracellular immunoglobulin variable (IgV) domain, a type I transmembrane domain, and an intracellular domain containing both an ITIM motif and an Ig-tail tyrosine-like motif (ITT). TIGIT is expressed on activated conventional αβ T cells, as well as on memory T cells, regulatory T cells (Tregs), follicular helper T cells, and NKT cells. By activating inhibitory receptors within T cells, natural killer (NK) cells, and Tregs, TIGIT has emerged as a promising target for immunotherapy.

The ligands for TIGIT 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 ligand binding. TIGIT effectively blocks the binding of CD155 to either CD96 or CD226, thereby providing further evidence 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.

CD155 is primarily expressed on the surface of dendritic cells (DCs), T cells, B cells, and macrophages; it is also expressed to varying degrees in non-hematopoietic tissues, such as the kidneys, the nervous system, and the intestines. Additionally, CD155 has been reported to be highly expressed in various human malignancies, including melanoma, pancreatic cancer, colon cancer, lung adenocarcinoma, and glioblastoma. As a cell adhesion molecule, CD155 influences cellular proliferation, migration, invasion, and adhesion through tumor-associated signaling pathways.

The TIGIT NFAT-Luc Jurkat reporter gene drug target model accurately simulates the in vivo signal transduction process of TIGIT.  The principle is illustrated in the figure below.

Figure 1. Schematic Diagram of the TIGIT NFAT-Luc Jurkat Cell Model

Figure 2.Recombinant TIGIT/NFAT-Luc/Jurkat stably expressing TIGIT.

Figure 3. Dose Response of TIGIT Monoclonal Antibody in TIGIT NFAT-Luc Jurkat(C26) with CD155/TCR activator-CHO.

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.