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 T cells play a central role in cell-mediated immunity, mediating long-lasting, antigen-specific, effector, and memory responses. They serve as targets for various immunotherapies, including checkpoint inhibitors, bispecific T-cell engagers, and immune agonists. Additionally, T cells themselves have been explored as therapies for hematologic malignancies through chimeric antigen receptor addition or autologous adoptive transfer. In recent years, diverse immunotherapeutic strategies aimed at inducing, enhancing, and/or engineering T-cell responses have emerged as promising approaches for treating diseases such as cancer and autoimmune disorders.

 The CD8 molecule, encoded by the CD8 gene, is a critical co-receptor on the surface of T cells, primarily expressed on cytotoxic T lymphocytes (CTLs). It forms a heterodimer through disulfide bonds connecting the α (CD8α) and β (CD8β) subunits or exists as an α-α homodimer. The core function of CD8 is to serve as an auxiliary molecule for the T cell receptor (TCR), enhancing the sensitivity of TCR to antigen-MHC complexes by binding to major histocompatibility complex class I (MHC I) via its extracellular segment. This activates the cytotoxic function of T cells, enabling precise elimination of virus-infected or tumor cells.

The CD8+ NFAT-Luc Jurkat reporter gene drug target model effectively mimics the in vivo CD8α and CD8β subunits as well as the downstream NFAT signaling pathway reporter gene, with the underlying mechanism illustrated in the figure below.

Figure 1. Schematic Diagram of the Jurkat E6.1 Human CD8+ NFAT-Luc Cell 

Figure 1.Recombinant CD8+ NFAT-Luc Jurkat  stably expressing CD8a&CD8b.

Figure 2. Dose Response of CD3 Antibody in CD8+ NFAT-Luc Jurkat(C40).

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.