IL-21 Reporter HEK 293 Cells

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Interleukin-21 Reporter Cells

Signaling pathway in HEK-Blue™ IL-21 cells

HEK-Blue™ IL-21 cells were engineered from the human embryonic kidney HEK 293 cell line to detect bioactive interleukin 21 (IL-21) by monitoring the activation of the JAK/STAT3 pathway. They can also be used for screening antibodies or small molecule inhibitors targeting the IL-21 pathway.

IL-21 is a secreted cytokine that plays an essential role in promoting the proliferation and development of T cell subsets and inducing B cell generation and differentiation into plasma cells [1]. 

 More details

Cell line description

HEK-Blue™ IL-21 cells were generated by stable transfection with the genes encoding the IL-21Rα and IL-2Rγ chains of human IL-21 receptor, JAK3, STAT3, and a STAT3-inducible secreted embryonic alkaline phosphatase (SEAP) reporter. The binding of IL-21 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-21 cells detect human (h) and murine (m) IL-21. Of note, they also respond to hIFN-α and hIFN-β, but not hIFN-γ (see figures).

Key Features

• Fully functional IL-21 signaling pathway
• Readily assessable STAT3-inducible SEAP reporter activity
• Strong response to human (h) and mouse (m) IL-21

Applications

• Detection and quantification of hIL-21 and mIL-21 activity
• Screening of anti-IL-21 and anti-IL-21 receptor antibodies
• Screening of small molecule inhibitors of the IL-21 pathway

Reference:

1. Spolski R. et al., 2014. Interleukin-21: a double-edged sword with therapeutic potential. Nat Rev Drug Discov. 13(5):379-95.

Figures

Dose-response of HEK-Blue™ IL-21 cells to recombinant IL-21 cytokines. Cells were stimulated with increasing concentrations of recombinant human (h)IL-21 or murine (m)IL-21. After overnight incubation, the NF-κB-induced SEAP activity was determined using QUANTI-Blue™ Solution, a SEAP detection reagent. Data are shown as optical density (OD) at 630 nm (mean ± SEM).

Cytokine response profile of HEK-Blue™ IL-21 cells. Cells were stimulated with various human and murine recombinant cytokines: hIFN-a (1000 U/ml), hIFN-b (1000 U/ml), hIFN-g (10 n/ml), hIL-21 (10 ng/ml), mIL-21 (10 ng/ml), hIL-2 (10 ng/ml), hIL-4 (10 ng/ml), hIL-6 (10 ng/ml), hIL-7 (10 ng/ ml), hIL-9 (10 ng/ml), IL-10 (10 ng/ml), hIL-15 (10 ng/ml), hIL-22 (10 ng/ml), and hIL-27 (10 ng/ml). After overnight incubation, the NF-κB-induced SEAP activity was determined using QUANTIBlue™ Solution. Data are shown as optical density (OD) at 630 nm (mean ± SEM).

Specifications

Antibiotic resistance: Blasticidin, Hygromycin B, Puromycin, Zeocin®

Growth medium: DMEM, 4.5 g/l glucose, 2 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-21 and murine IL-21

Detection range:

• Detection range for human IL-21: 0.1 - 100 ng/ml 
• Detection range for murine IL-21: 1 - 100 ng/ml

Quality Control:

• SEAP reporter activity in response to IL-21 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 HEK-Blue™ Selection (250X concentrate)
• 1 ml of Puromycin (10 mg/ml)
• 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)

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

Details

Interleukin 21 (IL-21) is a cytokine mainly produced by CD4+ T cells and natural killer T (NKT) cells. It has pleiotropic actions on a wide range of immune (i.e. lymphoid and myeloid populations) and non-immune cell types (i.e. epithelial cells) [1,2]. IL-21 belongs to the common γ chain cytokine family. Its receptor comprises the IL-21Rα (CD360) and IL-2Rγ (CD132) chains and it signals through tyrosine kinases of the Janus family (JAK1 and JAK3) and signal transducer and transcription activators (STAT). STAT3 plays a major role in the biological actions of IL-21, and some functions have been also attributed and suggested for STAT1 and STAT5, respectively [1-3]. Furthermore, IL-21 facilitates the PI3K/AKT and MAPK pathways, which both contribute to IL-21 signal transmission [1-3]. IL-21 is referred to as a double-edged sword cytokine, with key roles in anti-tumor and anti-viral responses, but also in promoting the development of autoimmune diseases (e.g. systemic lupus erythematosus and rheumatoid arthritis) [1-3]. Recombinant IL-21 as well as anti-IL-21 monoclonal antibodies are being investigated for therapeutic use in cancer and autoimmune diseases [1-3].

1. Spolski R. et al., 2014. Interleukin-21: a double-edged sword with therapeutic potential. Nat Rev Drug Discov. 13(5):379-95.
2. Leonard WJ. et al., 2016. IL-21 signaling in immunity. F1000Res. doi: 10.12688/f1000research.7634.1.
3. Long D. et al., 2019. Clinical significance and immunobiology of IL-21 in autoimmunity. J Autoimmunity. 99:1-14. 

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