IL-2 & IL-15 Reporter HEK 293 Cells expressing CD25, CD122, & CD132

Cytokine offer: This product includes a free vial of recombinant human IL-2 or IL-15 – select above! View all InvivoGen's Cytokine Bioassays.

Human/mouse IL-2 and human IL-15 Reporter Cells

Signaling pathway in HEK-Blue™-IL-2/IL-15 cells

HEK-Blue™ IL-2/IL-15 cells are designed to monitor human IL-2 and IL-15-induced STAT5 stimulation or inhibition through SEAP detection. This colorimetric bioassay can be used for screening activatory molecules, such as engineered cytokines, or inhibitory molecules, such as neutralizing antibodies.

HEK-Blue™ IL-2/IL-15 cells strongly respond to recombinant human IL-2 and human IL-15, as well as mouse IL-2. The reliable and consistent performance of HEK-Blue™ IL-2/IL-15 cells makes them suitable for release assays of therapeutic molecules that inhibit cytokine signaling, such as Ordesekimab, a monoclonal antibody targeting IL-15 (see figures).

Key features

• Readily assessable STAT5-SEAP reporter activity
• Convenient readout using  QUANTI-Blue™ Solution
• High sensitivity to human (h) IL-2 and hIL-15 activity
• Stability guaranteed for 20 passages

Applications

• Therapeutic development
• Drug screening
• Release assay

IL-2 and IL-15 detection potency

IL-2 and IL-15 are closely related cytokines that belong to the IL-2 family and display important functions in the immune system [1]. They share the heterodimeric CD122 (IL-2Rβ)/CD132 (IL-2Rγ) receptor to deliver their signals within target cells. Their specificity of action is conferred by their α receptor chains, CD25 (IL-2Rα) and CD215 (IL-15Rα) [1].

 More details

InvivoGen also offers HEK-Blue™ CD122/132 cells, expressing the  CD122 (IL-2Rβ) and CD132 (IL-2Rγ) subunits,  showing a different detection range for mouse IL-2 and IL-15 (see table and figures). 

Figures

Dose-response of HEK-Blue™ IL-2/Il-15 cells to recombinant IL-2 and IL-15.

Cells were stimulated with increasing concentrations of E. coli-derived 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.

Cytokine response profile of HEK-Blue™ IL-2/Il-15 cells.

Cells were stimulated with various E. coli-derived 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.

Dose-dependent inhibition of HEK-Blue™ IL-2/IL-15 cell response using Ordesekimab biosimilar. Increasing concentrations of Anti-hIL-15-hIgG1 (0.1 ng/ml - 10 µg/ml) were incubated with recombinant human (h)IL-15 (30 pg/ml) for 1 h before the addition of HEK-Blue™ IL-2/IL-15 cells. After overnight incubation, SEAP activity in the cell culture supernatant was assessed using QUANTI-Blue™ Solution. Data are shown in percentage of activity (%) (mean ± SEM).

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

Cells were stimulated with increasing concentrations of E. coli-derived 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

Cell type: Epithelial

Tissue origin: Human Embryonic Kidney

Target: IL-2, IL-15

Specificity: Human, Mouse

Reporter gene: SEAP

Antibiotic resistance: Blasticidin, Hygromycin B, Puromycin, Zeocin®

Detection ranges:

• Detection range for human IL-2 & mouse IL-2: 0.03 - 100 ng/ml
• ​Detection range for human IL-15: 0.01 - 100 ng/ml

Growth medium: Complete DMEM (see TDS)

Growth properties: Adherent

Mycoplasma-free: Verified using Plasmotest™

Quality control: Each lot is functionally tested and validated.

Contents

HEK-Blue™ IL-2/Il-15 cells (hkb-il2-2/-15)

• 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).

HEK-Blue™ IL-2/Il-15 vial (hkb-il2-av)

• 1 vial containing 3-7 x 10⁶ cells

Shipped on dry ice (Europe, USA, Canada and some areas in Asia)

Notification:  Reference #hkb-il2-av can only be ordered together with references #hkb-hkb-il2-2 or #hkb-il2-15.

Details

Cell line description

HEK-Blue™ IL-2/IL-15 cells were generated by stable transfection of the human embryonic kidney HEK293 cell line with the genes encoding for the human CD25 (IL-2Rα), CD122 (IL-2Rβ), and CD132 (IL-2Rγ) subunits. They also feature a stable expression of the human JAK3 and STAT5 to obtain a fully functional IL-2/IL-15 signaling pathway, as well as a STAT5-inducible secreted embryonic alkaline phosphatase (SEAP) reporter. The binding of IL-2 or IL-15 to their receptors triggers a signaling cascade leading to the activation of STAT5 and the subsequent production of SEAP. This can be readily assessed in the supernatant using QUANTI-Blue™ Solution, a SEAP detection reagent.

HEK-Blue™ IL-2/IL-15 cells respond to human IL-2 and IL-15 and thus can be used for the screening of anti-hIL-2 and anti-hIL-15 antibodies. The addition of CD25 in HEK-Blue™ IL-2/IL-15 cells does not increase their sensitivity to human IL-2; however, it does increase their sensitivity to mouse IL-2 (see figures). HEK-Blue™ IL-2/IL-15 cells can be used to validate the functionality, toxicity, and variable dosage effects of human or mouse IL-2. However, this cell line does not respond to mouse IL-15.

IL-2 and IL-15 background

IL-2 and IL-15 are two closely related cytokines that belong to the IL-2 family and display important functions in the immune system [1]. They share the heterodimeric CD122 (IL-2Rβ)/CD132 (IL-2Rγ) receptor to deliver their signals within target cells. Their specificity of action is conferred by their α receptor chains, CD25 (IL-2Rα) and CD215 (IL-15Rα). The binding of IL-2 and IL-15 to their heterodimeric or heterotrimeric receptors activates a signaling cascade through tyrosine kinases of the Janus family (JAK1 and JAK3) and signal transducer and transcription activator 5 (STAT5) [1]. This results in the subsequent induction of proliferation and anti-apoptotic transcriptional program and can, if uncontrolled, potentially lead to malignant transformation [2]. IL-15 can sustain chronic inflammation, autoimmunity, and uncontrolled T cell proliferation, leading to T cell lymphoma development [1].

1. Sindaco P, et al., 2023. The role of interleukin-15 in the development and treatment of hematological malignancies. Front Immunol.14:1141208.

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