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Glucocorticoid-induced tumor necrosis factor receptor-related protein (GITR, also known as TNFRSF18 or CD357) belongs to the tumor necrosis factor receptor superfamily (TNFRSF) and is expressed on various immune cells, including effector T cells and regulatory T cells. GITR plays a crucial role in regulating the immune system, particularly in the activation of T cells and the function of regulatory T cells (Tregs).

The ligand for GITR is GITRL, which is typically expressed on antigen-presenting cells. Upon binding of GITR to GITRL, downstream signaling pathways are triggered, including the NF-κB, MAPK, and PI3K/AKT pathways, thereby promoting T cell survival, proliferation, and effector functions. Concurrently, GITR activation may also interfere with the inhibitory capacity of regulatory T cells, further lifting immune suppression.

 The GITR NFκB-Luc Jurkat reporter model effectively mimics the in vivo GITR signaling pathway; the mechanism is illustrated in the figure below.

Figure 1. Schematic diagram of the GITR NFκB-Luc Jurkat cell model

Figure 2. Recombinant GITR/NFκB-Luc/Jurkat stably expressing GITR.

Figure 3.  Detect Luciferase assay by Ultra Luciferase Detection Kit CBPH0001（we strongly suggest to purchase from Cobioer). Dose Response of Recombinant Human TNFSF18 in GITR-NFκB-Luc Jurkat(C1). Dose Response of GITR NFκB-Luc Jurkat(C1) Stimulated by GITRL 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.