IL-4 & IL-13 Reporter HEK 293 Cells

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IL-4 & IL-13 Reporter Cells

Signaling pathway in HEK-Blue™ IL-4/IL-13 cells

HEK-Blue™ IL-4/IL-13 cells were engineered from the human embryonic kidney HEK293 cell line to detect bioactive human interleukin-4 (IL-4) and IL-13 by monitoring the activation of the STAT6 pathway. In addition, these cells can be used for screening antibodies or small molecule inhibitors targeting the IL-4 or IL-13 pathway.

IL-4 and IL-13 are cytokines that play an important role in the regulation of inflammation and immune responses [1].

 More details

Cell line description

HEK-Blue™ IL-4/IL-13 cells were generated by stable transfection with the gene encoding human STAT6 to obtain a fully active STAT6 pathway. The other genes of the IL-4/IL-13 pathway are naturally expressed in these cells. These cells were further transfected with a STAT6-inducible secreted embryonic alkaline phosphatase (SEAP) reporter. The binding of IL-4 or IL-13 to their receptor triggers a signaling cascade leading to STAT6 activation 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-4/IL-13 cells detect human (h) IL-4 and hIL-13 (see figures). Of note, they do not respond to hIFN-α or hIFN-β (see figures).

Key features

• Fully functional IL-4/IL-13 signaling pathway
• Readily assessable STAT6-SEAP reporter activity
• Strong response to human (h) IL-4 and hIL-13
• No response to hIFN-α or hIFN-β

Applications

• Detection and quantification of human IL-4 and human IL-13 activity
• Screening of anti-IL-4, anti-IL4Rα, anti-IL-13, or anti-IL-13Rα1 antibodies
• Screening of small molecule inhibitors of the IL-4 or IL-13 pathway

Reference:

1. Kasaian MT. et al., 2013. An IL-4/IL-13 dual antagonist reduces lung inflammation, airway hyperresponsiveness, and IgE production in mice. Am J Respir Cell Mol Biol.49(1):37-46.

Figures

Stimulation of HEK-Blue™ IL-4/IL-13 cells by recombinant human IL-4 or IL-13. After a 24 h incubation, SEAP activity was assessed using QUANTI-Blue™ Solution and reading the optical density (O.D.) at 655 nm.

Ligand	EC50	Response Ratio
Recombinant hIL-4	~0.7 ng/ml	25
Recombinant hIL-13	~2.0 ng/ml	28

HEK-Blue™ IL-4/IL-13 cells were stimulated with various human recombinant cytokines: IFN-α (1000 IU/ml), IFN-β (1000 IU/ml), IFN-γ (1000 IU/ml), IL-1β (100 ng/ml), IL-4 (10 ng/ml), IL-6 (100 ng/ml), IL-13 (10 ng/ml), IL-18 (100 ng/ml), TGF-β (10 ng/ml), and TNF-α (100 ng/ml). After a 24 h incubation, SEAP activity was assessed using QUANTI-Blue™ Solution and reading the O.D. at 655 nm.

Specifications

Antibiotic resistance: Blasticidin, Zeocin®

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

Specificity: Detects human IL-4 and human IL-13

Detection range:

• Detection range for human IL-4: 0.1 - 10 ng/ml
• Detection range for human IL-13: 1 - 100 ng/ml

Quality Control:

• SEAP reporter activity in response to IL-4 and IL-13 is validated using functional assays.
• The stability for 20 passages following thawing is confirmed.
• These cells are tested for mycoplasma contamination. 

Contents

• 1 vial containing 3-7 x 10⁶ cells
• 1 ml of Blasticidin (10 mg/ml)
• 1 ml of Zeocin® (100 mg/ml)
• 1 ml of 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)

Details

The transcription factor STAT6 is activated primarily by two cytokines with overlapping biologic functions, IL-4 and IL-13 [1, 2].

It can also be activated by IFN-α in a cell-specific manner. In non-hematopoietic cells, IL-4 and IL-13 bind a receptor complex composed of the IL-4Ralpha and IL-13Ralpha1.

Upon ligand binding, the receptor complex activates the receptor-associated Janus kinases (JAK1 and Tyk2) leading to the recruitment of STAT6 and its phosphorylation.

Activated STAT6 forms homodimers that translocate to the nucleus where they bind the promoter of responsive genes inducing gene transcription.

In HEK-Blue™ IL-4/IL-13 cells stimulation with IL-4 or IL-13 activates the JAK/STAT6 pathway with the subsequent production of SEAP.

1. Kasaian MT. et al., 2013. An IL-4/IL-13 dual antagonist reduces lung inflammation, airway hyperresponsiveness, and IgE production in mice. Am J Respir Cell Mol Biol.49(1):37-46.
2. David M. et al., 2003. Functional characterization of IL-13 receptor a2 gene promoter: a critical role of the transcription factor STAT6 for regulated expression. Oncogene 22, 3386–3394.

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