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

Human IL-2 and  IL-15 Reporter Cells

Signaling pathway in HEK-Blue™-CD122/CD132 cells

HEK-Blue™ CD122/CD132 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™ CD122/CD132 cells strongly respond to recombinant human IL-2 and human IL-15. Moreover, they respond poorly to mouse (m) IL-2, but not to mIL-15 (see figures). The reliable and consistent performance of HEK-Blue™ CD122/CD132 cells makes them suitable for release assays of therapeutic molecules that inhibit cytokine signaling, such as monoclonal antibodies targeting IL-2 or IL-15. 

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
• Poor response to mouse (m) IL-2
• 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™ IL-2/IL-15 cells, expressing the CD25 (IL-2Rα), CD122 (IL-2Rβ), and CD132 (IL-2Rγ) subunits, showing a different detection range for human/mouse IL-2 and IL-15 (see table and figures). 

Figures

Dose-response of HEK-Blue™ CD122/CD132 cells to recombinant IL-2 and IL-15.
Cells were stimulated with increasing concentrations of E. coli-derived recombinant human IL-2 (hIL-2) or human IL-15 (hIL-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™ CD122/CD132 cells.
Cells were stimulated with various E. coli-derived human (h) or murine (m) recombinant cytokines; 1 ng/ml hIL-2, 1 ng/ml mIL-2, 0.1 ng/ml hIL-15, 1 ng/ml mIL-15, 10 ng/ml TNF-α, and 1x103 IU/ml IFN-α or 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.

Specifications

Cell type: Epithelial

Tissue origin: Human Embryonic Kidney

Target: IL-2, IL-15

Specificity: Human

Reporter gene: SEAP

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

Detection ranges:

• Detection range for human IL-2: 0.1 - 10 ng/ml
• ​Detection range for human IL-15: 0.03 - 10 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™ CD122/CD132 Cells (hkb-il2bg)

• 1 vial containing 3-7 x 10⁶ cells
• 1 ml Puromycin (10 mg/ml)
• 2 x 1 ml HEK-Blue™ 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™ CD122/CD132 vial (hkb-il2bg-av)

• 1 vial containing 3-7 x 10⁶ cells

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

Notification:  The reference #hkb-il2bg-av can only be ordered together with reference #hkb-il2bg.

Details

Cell line description

HEK-Blue™ CD122/CD132 cells were generated by stable transfection of the human embryonic kidney HEK293 cell line with the genes encoding for the human 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™ CD122/CD132 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 (see figures). HEK-Blue™ CD122/CD132 cells can be used to validate the functionality, toxicity, and variable dosage effects of human IL-2. This cell line is specific to human IL-2 and IL-15 and thus does not respond to mouse IL-2 or 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.

FAQ Cell Lines

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