IL-2 Reporter HEK 293 Cells

HEK-Blue™ IL-2 Cells signaling pathway

Human & Murine IL-2 (and Human IL-15) Reporter Cells

HEK-Blue™ IL-2 reporter cells have been specifically designed to detect bioactive human and murine interleukin-2 (IL-2) by monitoring the activation of the JAK-STAT pathway. These cells also detect bioactive human IL-15.

IL-2 is a secreted cytokine that plays an important role in immune activation and homeostasis [1, 2]. IL-2 exerts its action by binding to the heterotrimeric IL-2 receptor, consisting of CD25 (aka IL-2Rα), CD122 (IL-2Rβ), and CD132 (IL-2Rγ). This binding leads to the activation of a signaling cascade through tyrosine kinases of the Janus family (JAK1 and JAK3) and signal transducer and transcription activator 5 (STAT5).

IL-2 is closely related to IL-15; they share the heterodimeric CD122/CD132 receptor to deliver their signals within target cells. Indeed, HEK-Blue™ IL-2 cells which overexpress CD122 and CD132 also respond to human IL-15 and thus can be used for the screening of anti-hIL15 antibodies.

Interestingly, HEK-Blue™ IL-2 cells and HEK-Blue™ CD122/CD132 cells, another HEK-derived cell line overexpressing only CD122 and CD132 (and not CD25), respond similarly to human IL-2 and IL-15. The addition of CD25 in HEK-Blue™ IL-2 cells does not increase their sensitivity to human IL-2, however, it does increase their sensitivity to murine IL-2 (see figures). 

Cell line description:

HEK-Blue™ IL-2 cells were generated by stable transfection of the human embryonic kidney HEK 293 cell line with the human CD25 (IL-2Rα), CD122 (IL-2Rβ), and CD132 (IL-2Rγ) genes, along with the human JAK3 and STAT5 genes to obtain a fully functional IL-2 signaling pathway. In addition, a STAT5-inducible SEAP reporter gene was also introduced. Upon IL-2 stimulation, HEK-Blue™ IL-2 cells trigger the activation of STAT5 and the subsequent secretion of SEAP. The levels of STAT5-induced SEAP can be readily monitored using QUANTI-Blue™ Solution.
HEK-Blue™ IL-2 cells can be used to validate the functionality, toxicity, and variable dosage effects of human or murine IL-2.

Features of HEK-Blue™ IL-2 cells:

• Fully functional IL-2 signaling pathway
• Readily assessable SEAP reporter activity
• Functionally tested and guaranteed mycoplasma-free

Applications of HEK-Blue™ IL-2 cells:

• Detection of hIL-2, mIL-2, and hIL-15
• Screening of anti-IL-2 and anti-hIL-15 antibodies

Read our review on IL-2: The Activator and Controller

References:

1. Choudhry H. et al., 2018. Prospects of IL-2 in Cancer Immunotherapy. Biomed Res Int. 2018:9056173.
2. Boyman O. & Sprent J., 2012. The role of interleukin-2 during homeostasis and activation of the immune system. Nat Rev Immunol. 12(3):180-90.

Figures

Figure 1. Dose-response of HEK-Blue™ IL-2 cells to recombinant IL-2 and IL-15.

Cells were stimulated with increasing concentrations of recombinant human and murine IL-2 or human IL-15. After overnight incubation, the STAT5 response was determined using QUANTI‑Blue™ Solution, a SEAP detection reagent, and reading the optical density (OD) at 630 nm.

Figure 2. Cytokine response profile of HEK-Blue™ IL-2 cells.

Cells were stimulated with various human recombinant cytokines; 1 ng/ml IL-2 or IL-15 and 10 ng/ml IL‑1β, IL-4, IL-6, IL-18, TGF-β, TNF-α, IFN-γ, and 1x104 IU/ml IFN-α, IFN-β.
After overnight incubation, SEAP activity was assessed using QUANTI‑Blue™ Solution, a SEAP detection reagent, and reading the optical density (OD) at 630 nm.

Figure 3. Dose-response of HEK-Blue™ IL-2 and HEK-Blue™ CD122/CD132 cells to recombinant IL-2 and IL-15.

Cells were stimulated with increasing concentrations of recombinant human and murine IL-2, and human IL-15. After overnight incubation, the STAT5 response was determined using QUANTI‑Blue™ Solution, a SEAP detection reagent, and reading the optical density (OD) at 630 nm.

Specifications

Antibiotic resistance: blasticidin, hygromycin, puromycin, Zeocin™

Growth medium: DMEM, 4.5 g/l glucose, 2-4 mM L-glutamine, 10% (v/v) fetal bovine serum, 100 U/ml penicillin, 100 μg/ml streptomycin, 100 μg/ml Normocin™

Guaranteed mycoplasma-free

Specificity: human & murine IL-2; human IL-15

Detection ranges:

• 0.03 - 100 ng/ml for human & murine IL-2
• ​0.01 - 100 ng/ml for human IL-15

Contents

• 1 vial containing 3-7 x 10⁶ cells
• 1 ml puromycin (10 mg/ml)
• 2 x 1 ml HEK-Blue™ CLR Selection (250X concentrate); a solution containing several selection antibiotics.
• 1 ml Normocin™ (50 mg/ml)
• 1 ml of QB reagent and 1 ml of QB buffer (sufficient to prepare 100 ml of QUANTI-Blue™ Solution, a SEAP detection reagent).

Shipped on dry ice (Europe, USA & Canada)

FAQ

Any questions about our cell lines ? Visit our frequently asked questions page