IL-23 Reporter HEK 293 Cells

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Interleukin-23 Reporter Cells

Signaling pathway in HEK-Blue™ IL-23 cells

HEK-Blue™ IL-23 cells were engineered from the human embryonic kidney HEK 293 cell line to detect bioactive human and mouse interleukin-23 (IL-23) by monitoring the activation of the JAK/STAT3 pathway. They can also be used for screening antibodies or small molecule inhibitors targeting the IL-23 pathway.

IL-23 is a key cytokine in the promotion of chronic inflammation and plays a crucial role in the development and maintenance of T helper 17 (Th17) cells [1-3].

 More details

Cell line description

HEK-Blue™ IL-23 cells were generated by stable transfection with the genes encoding for the human IL-23 receptor (IL-23R and IL-12Rβ1 chains), as well as STAT3 to obtain a fully functional IL-23 signaling pathway. In addition, they express a STAT3-inducible secreted embryonic alkaline phosphatase (SEAP) reporter. The binding of IL-23 to its receptor triggers a signaling cascade leading to the activation of STAT3 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-23 cells detect human (h) and murine (m) IL-23 (see figures). Of note, these cells also respond to h/mIL‑27, another STAT3-signaling cytokine. Furthermore, due to the endogenous expression of the interferon-α/β receptor (IFNAR), these cells respond to hIFN-β (see figures).

Key features

• Fully functional IL-23 signaling pathway
• Readily assessable STAT3-inducible SEAP reporter activity
• Strong response to human (h) and mouse (m) IL-23

Applications

• Detection and quantification of hIL-23 and mIL-23 activity
• Screening of anti-IL-23 and anti-IL-23 receptor antibodies
• Screening of small molecule inhibitors of the IL-23 pathway

References:

1. Teng MW. et al., 2015. IL-23 and IL-23 cytokines: from discovery to targeted therapies for immune-mediated inflammatory diseases. Nat Med. 21(7):719‑29.
2. Vignali DA. & Kuchroo VK., 2012. IL-12 family cytokines: immunological playmakers. Nat Immunol. 13(8):722-8.
3. Duvallet E. et al., 2011. Interleukin-23: a key cytokine in inflammatory diseases. Ann Med. 43(7):503-11.

Figures

Dose-response of HEK-Blue™ IL-23 cells to recombinant IL-23.

Cells were stimulated with increasing concentrations of recombinant human and murine IL-23. After overnight incubation, the STAT3 response was determined using QUANTI-Blue™, a SEAP detection reagent, and reading the optical density (OD) at 630 nm. EC₅₀ values are shown as mean ± SD.

Cytokine response profile of HEK-Blue™ IL-23 cells.

Cells were stimulated with various human and murine recombinant cytokines; 3 ng/ml of hIL-23, mIL-23, hIL-27, mIL-27, hIL-12, mIL-12, hIL-4, hIL-6, hTNF-α, and 1x10³ IU/ml hIFN-β. After overnight incubation, SEAP activity was assessed using QUANTI-Blue™. Of note, at high concentrations hIL-6 and hTNF-α may induce a weak response, while hIL-12 and mIL-12 do not induce a response even at high concentrations.

Specifications

Antibiotic resistance: Blasticidin, Hygromycin B, 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®

Specificity: Detects human IL-23 and mouse IL-23

Detection range:

• Detection range for human IL-23: 100 pg/ml - 10 ng/ml
• Detection range for murine IL-23: 100 pg/ml - 10 ng/ml

Quality Control:

• SEAP reporter activity in response to IL-23 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
• 2 x 1 ml of HEK-Blue Selection (250X concentrate)
• 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

Interleukin 23 (IL-23), a member of the IL-12 family, is a heterodimeric cytokine composed of the IL-12p40 and IL-23p19 subunits.

IL-23 plays a crucial role in the development and maintenance of T helper 17 cells [1]. It acts by binding to a receptor complex consisting of the IL-12 receptor β1 (IL-12Rβ1) and the IL-23 receptor (IL-23R). Upon binding, IL-23 triggers a signaling pathway involving tyrosine kinase 2 (TyK2) and Janus kinase 2 (JAK2) leading to the activation of signal transducer and activator of transcription 3 (STAT3).

IL-23 has been identified as a central cytokine in autoimmunity and a highly promising treatment target for inflammatory diseases [1-3].

1. Teng MW. et al., 2015. IL-23 and IL-23 cytokines: from discovery to targeted therapies for immune-mediated inflammatory diseases. Nat Med. 21(7):719‑29.
2. Vignali DA. & Kuchroo VK., 2012. IL-12 family cytokines: immunological playmakers. Nat Immunol. 13(8):722-8.
3. Duvallet E. et al., 2011. Interleukin-23: a key cytokine in inflammatory diseases. Ann Med. 43(7):503-11.

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