IL-2 & IL-15 Reporter HEK 293 Cells

Notification:  References #hkb-il2-av and #hkb-il2bg-av can only be ordered together with references #hkb-hkb-il2 and #hkb-il2bg, respectively.

IL-2 and  IL-15 Reporter Cells

IL-2 and IL-15 sensing in HEK-Blue™-derived cells

HEK-Blue™ IL-2 and HEK-Blue™ CD122/CD132 cells were engineered from the human embryonic kidney HEK 293 cell line to detect bioactive interleukin-2 (IL-2) and interleukin-15 (IL-15) by monitoring the activation of the JAK/STAT5 pathway.

 –  HEK-Blue™ IL-2 cells for detection of human and murine IL-2 (and human IL-15) 
 –  HEK-Blue™ CD122/CD132 cells for detection of human IL-2 and human IL-15

In addition, these cells can be used for screening antibodies or small molecule inhibitors targeting the IL-2 or IL-15 pathway. 

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). The binding of IL-2 and IL-15 to their heterodimeric or heterotrimeric receptors 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) [1].

Cell line description

HEK-Blue™ IL-2 and HEK-Blue™ CD122/CD132 cells were generated by stable transfection with the genes encoding for the human CD25 (IL-2Rα), CD122 (IL-2Rβ), and CD132 (IL-2Rγ) subunits, or both CD122 (IL-2Rβ) and CD132 (IL-2Rγ) subunits, respectively. Both cell lines also feature a stable expression of the human JAK3 and STAT5 to obtain a fully functional IL-2 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 cells and HEK-Blue™ CD122/CD132 cells respond similarly to human IL-2 and IL-15 and thus can be used for the screening of anti-hIL2 and anti-hIL-15 antibodies. 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). HEK-Blue™ IL-2 cells can be used to validate the functionality, toxicity, and variable dosage effects of human or murine IL-2. HEK-Blue™ IL-2 and HEK-Blue CD122/CD132 cells do not respond to murine IL-15.

Key features

• Fully functional IL-2 and IL-15 signaling pathways
• Readily assessable STAT5-inducible SEAP reporter activity

Applications

IL-2 and IL-15 detection potency

• Detection and quantification of hIL-2, mIL-2, and hIL-15 activity using HEK-Blue™ IL-2 cells
• Detection and quantification of hIL-2 and hIL-15 activity using HEK-Blue™ CD122/CD132 cells
• Screening of anti-IL-2 or -IL-15 and anti-IL-2 or -IL-15 receptor antibodies
• Screening of small molecule inhibitors of the IL-2 or IL-15 pathway

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

References:

1. Morris R. et al., 2018. The molecular details of cytokine signaling via the JAK/STAT pathway. Protein Science. 27(12):1984-2009.

Figures

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.

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.

Dose-response of HEK-Blue™ CD122/CD132 cells to recombinant IL-2 and IL-15.
Cells were stimulated with increasing concentrations of 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 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.

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

HEK-Blue™ IL-2 cells

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

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

Specificity: Detects human IL-2, murine IL-2; and human IL-15

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

Quality Control:

• SEAP reporter activity in response to IL-2 and IL-15 is validated using functional assays.
• The stability for 20 passages following thawing is confirmed.
• These cells are tested for mycoplasma contamination. 

HEK-Blue™ CD122/CD132 cells

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

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

Specificity: Detects human IL-2 and human IL-15

Detection ranges:

• Detection range for human IL-2: 0.1 - 10 ng/ml
• ​Detection range for human IL-15: 0.03 - 10 ng/ml

Quality Control:

• SEAP reporter activity in response to IL-2 and IL-15 is validated using functional assays.
• The stability for 20 passages following thawing is confirmed.
• These cells are tested for mycoplasma contamination. 

Contents

Note: Each cell line is sold separately.

HEK-Blue™ IL-2 cells

• 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™ CD122/CD132 cells

• 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).

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

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