IL-22 Reporter HEK 293 Cells

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IL-22 Reporter Cells

HEK-Blue™ IL-22 Cells signaling pathway

HEK-Blue™ IL-22 cells were engineered from the human embryonic kidney HEK293 cell line to detect bioactive human and murine interleukin 22 (IL-22) by monitoring the activation of the STAT3 pathway. These cells can also be used for screening anti-IL-22 antibodies. IL-22 is a key regulator of immunity and inflammation at mucosal surfaces where it helps in maintaining barrier integrity [1-3]. 

 More details

Cell line description:

HEK-Blue™ IL-22 cells were generated by stable transfection with genes encoding for the human IL-22 receptor (IL-22R1 and IL-10Rβ chains),  STAT3, and a STAT3-inducible secreted embryonic alkaline phosphatase (SEAP) reporter. The binding of IL-22 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-22 cells detect human (h) and murine (m) IL-22. Of note, as HEK293 and HEK-Blue™ IL-22 cells endogenously express the interferon-α/β receptor, they display a strong response to hIFN-β (see figures).

Key features:

• Fully functional IL-22 signaling pathway
• Readily assessable STAT3-inducible SEAP reporter activity

Applications:

• Detection of both human and murine IL-22
• Screening of anti-IL-22 antibodies

References:

1. Wang J.et al., 2018. Aryl hydrocarbon receptor/IL-22/Stat3 signaling pathway is involved in the modulation of intestinal mucosa antimicrobial molecules by commensal microbiota in mice. Innate Immun. 24(5):297-306.
2. Foxall R.B. et al.., 2016. Profile of interleukin-22 in gut mucosal health and disease. IJICMR. 8:1-11. 
3. Park J.H. et al., 2017. There Is a Gap in Our Knowledge. Immunohorizons. 2(6):198-207. 

Figures

Dose-response of HEK-Blue™ IL-22 cells to recombinant IL-22.

Cells were stimulated with increasing concentrations of recombinant human or murine IL-22. After overnight incubation, the STAT3 response was determined using QUANTI‑Blue™ Solution, a SEAP detection reagent, and reading the optical density (OD) at 630 nm. EC50 values are shown as mean ± SD.

Cytokine response of HEK-Blue™ IL-22 cells.

Cells were stimulated with various human and murine recombinant cytokines: 1 ng/ml of hIL‑22, mIL-22, hIL‑2, hIL-6, hIL-10, hIL-27, hIL-28A, hIL-28B, hIL-29, hTNF-α, 1x10³ hIFN-β. After overnight incubation, SEAP activity was assessed using QUANTI‑Blue™ Solution. 

Dose-dependent inhibition of HEK‑Blue™ IL-22 cellular response using a neutralizing antibody against the IL-22 receptor (IL‑22R).

The antibody was incubated with HEK‑Blue™ IL-22 cells for 30 minutes prior to the addition of hIL-22 (10 ng/ml). After overnight incubation, SEAP activity in the cell culture supernatant was assessed using QUANTI‑Blue™ Solution. Data represent % inhibition of reporter activity without the anti‑IL-22R antibody.

Specifications

Antibiotic resistance: Blasticidin, Puromycin, 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™

Guaranteed mycoplasma-free

Specificity: human and mouse IL-22

Detection range:

• 0.03 - 10 ng/ml for human IL-22
• 0.1 - 10 ng/ml for murine IL-22

Contents

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

Interleukin 22 (IL-22) is a key regulator of immunity and inflammation at mucosal surfaces where it helps in maintaining barrier integrity [1-3]. IL-22 production can be triggered by a variety of pathogen-associated molecular patterns (PAMPs). Notably, it can be induced directly by Toll-like receptor 2 (TLR2) activation in response to bacterial-derived agonists, or indirectly via IL-23 in response to aryl-hydrocarbon receptor (AhR) ligands [1, 2]. IL-22 is implicated in a number of pathologies including autoimmune diseases and cancer [3, 4].  

IL-22 exerts its biological effect upon binding to its receptor, which comprises two subunits: IL-22R1 and IL-10Rβ. Upon binding, IL-22 triggers a signaling pathway involving tyrosine kinase 2 (TyK2) and Janus kinase 1 (JAK1) leading to the activation of signal transducer and activator of transcription 3 (STAT3). 

1. Wang J.et al., 2018. Aryl hydrocarbon receptor/IL-22/Stat3 signaling pathway is involved in the modulation of intestinal mucosa antimicrobial molecules by commensal microbiota in mice. Innate Immun. 24(5):297-306.
2. Foxall R.B. et al.., 2016. Profile of interleukin-22 in gut mucosal health and disease. IJICMR. 8:1-11. 
3. Park J.H. et al., 2017. There Is a Gap in Our Knowledge. Immunohorizons. 2(6):198-207. 
4. Hernandez P. et al., 2018. A catch-22: Interleukin-22 and cancer. Eur J Immunol. 48(1):15-31.

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