IL-31 Reporter HEK 293 Cells

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Interleukin-31 Reporter Cells

Signaling pathway in HEK-Blue™ IL-31 cells

HEK-Blue™ IL-31 cells were engineered from the human embryonic kidney HEK 293 cell line to detect bioactive interleukin-31 (IL-31) by monitoring the activation of the JAK/STAT5 pathway. They can also be used for screening antibodies or small molecule inhibitors targeting the IL-31 pathway.

IL-31 is a secreted cytokine that seems to be strongly involved in chronic inflammation in different skin diseases, including atopic dermatitis and allergic contact dermatitis [1].

 More details

Cell line description

HEK-Blue™ IL-31 cells were generated by stable transfection with the genes encoding for the human IL-31 receptor (IL-31Rα and human OSMRβ (oncostatin M receptor β) chains), STAT5b, and a STAT5-inducible secreted embryonic alkaline phosphatase (SEAP) reporter. The binding of IL-31 to its receptor 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-31 cells detect human IL-31 but not murine IL-31 (see figures). Of note, they also respond to hIFN-γ. However, they do not respond to hIFN-α and hIFN-β (see figures).

Key features

• Fully functional IL-31 signaling pathway
• Readily assessable STAT5-inducible SEAP reporter activity
• Strong response to human (h) IL-31
• No response to mouse (m) IL-31

Applications

• Detection and quantification of hIL-31 activity
• Screening of anti-IL-31 and anti-IL-31 receptor antibodies
• Screening of small molecule inhibitors of the IL-31 pathway

Reference:

1. Datsi A, et al., 2021. Interleukin-31: The "itchy" cytokine in inflammation and therapy. Allergy. 76: 2982-2997.

Figures

Validation of the expression of human IL-31 receptor chains by HEK-Blue™ IL-31 cells. 5 x 10⁵ cells were incubated with either a (A) PE-conjugated isotype control (blue) or a PE-conjugated Anti-hIL-31Rα mAb (red), or (B) APC-conjugated isotype control (blue) or a APC-conjugated Anti-hOSMR mAb (red) for 30 minutes. The binding affinity was then measured using flow cytometry.

Dose-response of HEK-Blue™ IL-31 cells to recombinant IL-31. Cells were stimulated with increasing concentrations of recombinant human IL-31 (hIL-31) and murine IL-31 (mIL-31). After overnight incubation, the STAT5 response was determined using QUANTI‑Blue™ Solution, a SEAP detection reagent. The optical density (OD) at 630 nm is shown as mean ± SEM.

Dose-dependent inhibition of HEK‑Blue™ IL-31 cellular response using a neutralizing antibody against IL-31Rα. An increasing concentration of a neutralizing anti-IL-31RA antibody was incubated with HEK‑Blue™ IL-31 cells for 3 hours before the addition of recombinant human IL-31 (0.03 ng/ml). After overnight incubation, SEAP activity in the cell culture supernatant was assessed using QUANTI-Blue™. Data are shown as a percentage (%) of activity.

Response of HEK-Blue™ IL-31 cells to a panel of cytokines. Cells were stimulated with various human and murine recombinant cytokines: 1 ng/ml of hIL-31, 100 ng/ml mIL-31, 10 ng/ml hIL-6, 100 ng/ml hIL-11, IL-27, hIFN-γ, and 1000 U/ml hIFN-α or IFN-β. After overnight incubation, SEAP activity was assessed using QUANTI‑Blue™. The OD at 630 nm is shown as mean ± SEM.

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-31

Detection range:

• Detection range for human IL-31: 1 pg/ml - 1 ng/ml

Quality Control:

• The expression of human IL-31Rα and OSMRβ is assessed by flow cytometry.
• SEAP reporter activity in response to IL-31 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 Normocin® (50 mg/ml)
• 2 x 1 ml of HEK-Blue Selection (250X)
• 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 cytokine interleukin 31 (IL-31) belongs to the Type I/II cytokine receptor family, and more specifically to the IL-6/gp130 family. IL-31 signals through a heterodimeric receptor comprised of the gp130-like subunit IL-31Rα and the OSMRb (oncostatin M receptor) subunit. The binding of IL-31 to its receptor triggers activation of the JAK/STAT, Akt/PI3K, and MAPK signaling pathways [1,2]. IL-31 receptor complex is expressed in epithelial, neuronal as well as immune cells. Little is known about the physiological function of IL-31. It is mainly secreted by IL-4-activated T cells, thereby contributing to Th2-weighted inflammation [1,2]. Several reports indicate that IL-31 is implicated in disorders such as atopic dermatitis, asthma, and inflammatory bowel disease [1,2]. Of note, IL-31 appears a critical cytokine involved in neuro-immune communication, through the mediation of inflammatory itch [1,2]. Targeting the IL-31 pathway is a promising therapeutic option for dermatologic and non-dermatologic diseases with enhanced IL-31 expression [1,2].

​References:

1. Bagci I.S. and Ruzicka, T., 2018. IL-31: A new player in dermatology and beyond. J. Allergy Clin Immunol. 141(3):858-866.
2. Datsi A., et al., 2021. Interleukin-31: The “itchy” cytokine in inflammation and therapy. Allergy. 76:2982-97.

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