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NKG2A is a member of the C-type lectin superfamily, specifically a single-pass type II transmembrane glycoprotein comprising an intracellular domain, a transmembrane domain, and an extracellular lectin-like domain. The intracellular domain contains two ITIMs (immunoreceptor tyrosine-based inhibitory motifs) that participate in inhibitory signal transduction. NKG2A serves as a critical immune checkpoint for natural killer (NK) cells and CD8+ T cells. NKG2A is expressed as a heterodimer with CD94—which also belongs to the C-type lectin superfamily—and the ligand for this NKG2A/CD94 heterodimeric receptor is a non-classical MHC Class I molecule. Upon binding, the NKG2A/CD94 complex transmits inhibitory signals to NK cells and CD8+ T cells; consequently, blocking NKG2A effectively unleashes the cytotoxic functions of these lymphocytes.

Binding of the NKG2A/CD94 receptor to the peptide-presenting molecule HLA-E triggers the phosphorylation of the ITIMs within NKG2A. These phosphorylated ITIMs are responsible for recruiting and activating the intracellular phosphatases SHP-1 and SHP-2, thereby suppressing the activating signals generated by receptors such as the T-cell receptor (TCR) and NKG2D. In contrast to classical HLA molecules—which are often lost—the expression of HLA-E is frequently upregulated in tumor cells. The interaction between NKG2A and HLA-E contributes to tumor immune evasion, and NKG2A-mediated mechanisms are currently being leveraged to develop potential anti-tumor therapeutic strategies. Furthermore, disrupting the interaction between NKG2A and its ligand has been demonstrated to effectively enhance anti-tumor immune responses. The expression level of NKG2A correlates with disease severity in patients with COVID-19. Additionally, NKG2A is implicated in the pathological processes of various other immune-mediated conditions, including autoimmune diseases, inflammatory disorders, parasitic infections, and transplant rejection. These findings suggest that NKG2A represents a novel therapeutic target for a wide spectrum of immune-mediated diseases. Furthermore, increasing evidence indicates that NKG2A also plays a significant role in other immune-related diseases, including viral infections, autoimmune diseases, inflammatory diseases, parasitic infections, and transplant rejection.

The Jurkat E6.1 Human NKG2A Effector Reporter Cell Model—effectively simulates the signal transduction process of NKG2A *in vivo*. The underlying principle is illustrated in the figure below.

Figure 1. Schematic Diagram of the Jurkat E6.1 Human NKG2A Effector Reporter Cell Model

Figure 2. Recombinant NKG2A Effector Reporter Cell expressing NKG2A and CD94.

Figure 3. Dose Response of NKG2A Blocking Ab in NKG2A Effector Reporter Cell(C56) With HLA-E 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.