IL-31 Reporter HEK 293 Cells

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IL-31 responsive STAT5-SEAP reporter assay

Signaling pathway in HEK-Blue™ IL-31 cells

HEK-Blue™ IL-31 cells are designed to monitor human IL-31-induced STAT5 stimulation or inhibition. This colorimetric bioassay can be used for screening activatory molecules, such as engineered cytokines, or inhibitory molecules, such as neutralizing antibodies.

HEK-Blue™ IL-31 cells respond specifically to recombinant human IL-31. The reliable and consistent performance of HEK-Blue™ IL-31 cells makes them suitable for release assays of therapeutic molecules that inhibit IL-31 signaling, such as Nemolizumab, a monoclonal antibody targeting the IL-31Rα subunit of the IL-31 receptor (see figures).

Key features

• Readily assessable STAT5-SEAP reporter activity
• Convenient readout using  QUANTI-Blue™ Solution
• High sensitivity to human (h) IL-31 activity
• Stability guaranteed for 20 passages

Applications

• Therapeutic development
• Drug screening
• Release assay

Interleukin-31 (IL-31) is a cytokine that plays a role in early protective inflammatory processes, likely regulating alarmins in damaged mucosal tissues. IL-31 is strongly involved in chronic inflammation in different skin diseases, including atopic dermatitis and allergic contact dermatitis.

 More details

All InvivoGen products are for internal research use only, and not for human or veterinary use.

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 650 nm is shown as mean ± SEM.

Dose-dependent inhibition of HEK‑Blue™ IL-31 cellular response using Nemolizumab biosimilar. Increasing concentrations of Anti-hIL-31R-hIgG2 (1 ng/ml - 1 µg/ml) were incubated with HEK‑Blue™ IL-31 cells for 3 hours before the addition of recombinant human IL-31 (10 pg/ml). After overnight incubation, SEAP activity in the cell culture supernatant was assessed using QUANTI-Blue™. Data are shown in percentage of activity (mean ± SEM).

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 650 nm is shown as mean ± SEM.

Specifications

Cell type: Epithelial

Tissue origin: Human Embryonic Kidney

Target: IL-31

Specificity: Human

Reporter gene: SEAP

Antibiotic resistance: Blasticidin, Hygromycin B, Zeocin®

Detection range: 2 pg/ml - 1 ng/ml (hIL-31)

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™ IL-31 Cells (hkb-il31)

• 1 vial containing 3-7 x 10⁶ cells
• 2 x 1 ml HEK-Blue™ Selection (250X concentrate)
• 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™ IL-31 vial (hkb-il31-av)

• 1 vial containing 3-7 x 10⁶ cells

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

Notification:  Reference #hkb-il31-av can only be ordered together with reference #hkb-il31.

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.
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).

IL-31 background

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 to be 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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