IL-21 Reporter HEK 293 Cells

HEK-Blue™ IL-21 Cells signaling pathway

Interleukin-21 Reporter 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.  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]. 

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Description:

HEK-Blue™ IL-21 cells were generated by stable overexpression of the genes encoding the alpha chain of human IL-21 receptor (IL-21Rα), IL-2Rγ, JAK3, STAT3, and a STAT3-inducible secreted embryonic alkaline phosphatase (SEAP) reporter. STAT3-dependent SEAP activity is readily assessable in the supernatant using QUANTI-Blue™ Solution, a detection reagent. Of note, HEK-Blue™ IL-21 cells also respond to human type I IFNs (IFN-α/IFN-β), but not human type II interferons (IFN-γ; see figures).

Key Features:

• Fully functional IL-21 signaling pathway
• Readily assessable STAT3-inducible SEAP reporter activity

Applications:

• Detection of human and murine IL-21
• Screening of anti-IL-21 and anti-IL-21R antibodies

References:

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, 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™

Guaranteed mycoplasma-free

Specificity: human and murine IL-21

Detection ranges:

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

This product is covered by a Limited Use License (See Terms and Conditions).

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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