IL-19, IL-20, IL-22 & IL-24 Reporter HEK 293 Cells

Notification:  References #hkb-il1920-av and #hkb-il20-av can only be ordered together with references #hkb-il1920 and #hkb-il20, respectively.

IL-19, IL-20, IL-22, and IL-24 sensing
in HEK-Blue™-derived cells

IL-19, IL-20 and  IL-24 Reporter Cells

HEK-Blue™ IL-19/IL-20 and HEK-Blue™ IL-20 reporter cells were engineered from the human embryonic kidney HEK 293 cell line to detect bioactive interleukin 19 (IL-19), IL-20, IL-22, and IL-24 by monitoring the activation of the JAK/STAT3 pathway:

— HEK-Blue™ IL-19/IL-20 cells expressing the IL-20 type I receptor for the detection of human and murine IL-19, IL-20, and IL-24 

— HEK-Blue™ IL-20 cells expressing the IL-20 type II receptor for the detection of human and murine IL-20, IL-22, and IL-24 

IL-19, IL-20, IL-22, and IL-24 are closely related cytokines that belong to the IL-20 subfamily and play an important role in the protection of barrier tissues. They share the heterodimeric IL-20 type I and type II receptors composed of the IL-20Rα and IL-20Rβ chains, and IL-22Rα1 and IL-20Rβ chains, respectively [1, 2].

 More details

Cell Line description:

HEK-Blue™ IL-19/IL-20 and HEK-Blue™ IL-20 cells were generated by stable transfection with the genes encoding for the human IL-20 type I receptor IL-20Rα and IL-20Rβ chains, or type II receptor IL-20Rβ and IL-22Rα chains, respectively. Both cell lines also feature a stable expression of human STAT3 to obtain a fully functional IL-19, IL-20, or IL-24 signaling pathway as well as a STAT3-inducible secreted embryonic alkaline phosphatase (SEAP) reporter. The binding of IL-19, IL-20, IL-22, or IL-24 to their 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-19/IL-20 cells, expressing the IL-20Rα and IL-20Rβ chains (type I receptor), respond to IL-19, IL-20, or IL-24 stimulation. They also respond to IL-27 and IFN-γ. They do not respond to IL-6, IL-10, IL-22, IL-26, and IFN-α/β (see figures).

— HEK-Blue™ IL-20 cells,  expressing the IL-22Rα and IL-20Rβ chains (type II receptor), respond to IL-20, IL-22, or IL-24 stimulation. They also respond to IL-27 and IFN-γ. They do not respond to IL-6, IL-10, IL-19, IL-26, and IFN-α/β (see figures).

Key features:

• Fully functional IL-19, IL-20, IL-22, and IL-24 signaling pathways
• Readily assessable STAT3-inducible SEAP reporter activity

Applications:

• Detection of human and murine IL-19, IL-20, and IL-24 using HEK-Blue™ IL-19/IL-20 cells
• Detection of human and murine IL-20, IL-22, and hIL-24 using HEK-Blue™ IL-20 cells
• Screening of anti-IL-19, anti-IL-20, anti-IL-22, and anti-IL-24 antibodies

References:

1. Ouyang W. et al., 2019. IL-10 family cytokines IL-10 and IL-22: from basic science to clinical translation. Immunity. 50:871-891.
2. Wei H. et al., 2019. Interleukin-10 family cytokines: Immunobiology and structure. Adv Exp Med Biol. 1172:79-96.

Figures

Cytokine response profile of HEK-Blue™ IL-19/IL-20 cells. Cells were stimulated with various human and murine recombinant cytokines: hIFN-α (1000 U/ml), hIFN-β (1000 U/ml), hIFN-γ (10 ng/ml), hIL-6 (10 ng/ml), hIL-10 (10 ng/ml), hIL-19 (10 ng/ml), mIL-19 (10 ng/ml), hIL-20 (10 ng/ml), mIL-20 (10 ng/ml), hIL-22 (10 ng/ml), mIL-22 (10 ng/ml), hIL-24 (10 ng/ml), mIL-24 (10 ng/ml),  hIL-26 (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).

Dose-response of HEK-Blue™ IL-19/IL-20 cells to recombinant cytokines. Cells were stimulated with increasing concentrations of recombinant (A) human or (B) murine IL-19, IL-20, IL-22, and IL-24. 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). 

Inhibition of hIL-24 signaling in HEK-Blue™ IL-19/IL-20 cells. Increasing concentrations of an anti-hIL-24 monoclonal antibody (mAb) were incubated for 30 minutes with the recombinant cytokine hIL-24 (3 ng/ml) prior to the addition of HEK-Blue™ IL-19/20 cells. After overnight incubation, SEAP activity in the cell culture supernatant was assessed using QUANTI-Blue™ Solution. Data are shown as optical density (OD) at 630 nm (mean ± SEM).

Cytokine response profile of HEK-Blue™ IL-20 cells. Cells were stimulated with various human and murine recombinant cytokines: hIFN-α (1000 U/ml), hIFN-β (1000 U/ml), hIFN-γ (10 ng/ml), hIL-6 (10 ng/ml), hIL-10 (10 ng/ml), hIL-19 (10 ng/ml), mIL-19 (10 ng/ml), hIL-20 (10 ng/ml), mIL-20 (10 ng/ml), hIL-22 (10 ng/ml), mIL-22 (10 ng/ml), hIL-24 (10 ng/ml), mIL-24 (10 ng/ml), hIL-26 (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).

Dose-response of HEK-Blue™ IL-20 cells to recombinant cytokines. Cells were stimulated with increasing concentrations of recombinant (A) human or (B) murine IL-19, IL-20, IL-22, and IL-24. 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). 

Inhibition of hIL-24 signaling in HEK-Blue™ IL-20 cells. Increasing concentrations of an anti-hIL-24 monoclonal antibody (mAb) were incubated for 30 minutes with the recombinant cytokine hIL-24 (3 ng/ml) prior to the addition of HEK-Blue™ IL-20 cells. After overnight incubation, SEAP activity in the cell culture supernatant was assessed using QUANTI-Blue™ Solution. Data are shown as optical density (OD) at 630 nm (mean ± SEM).

Specifications

HEK-Blue™ IL-19/IL-20 cells

Antibiotic resistance: Blasticidin, Puromycin, Zeocin®

Growth medium: DMEM, 4.5 g/l glucose, 2 mM L-glutamine, 10% (v/v) fetal bovine serum, 100 U/ml penicillin, 100 μg/ml streptomycin, 100 μg/ml Normocin™

Guaranteed mycoplasma-free

Specificity: Human & murine IL-19, IL-20, and IL-24

Detection ranges:

• 3 - 100 ng/ml for human & murine IL-19
• 1 - 100 ng/ml for human & murine IL-20
• 1 - 100 ng/ml for human & murine IL-24

HEK-Blue™ IL-20 cells

Antibiotic resistance: Blasticidin, Puromycin, Zeocin®

Growth medium: DMEM, 4.5 g/l glucose, 2 mM L-glutamine, 10% (v/v) fetal bovine serum, 100 U/ml penicillin, 100 μg/ml streptomycin, 100 μg/ml Normocin™

Guaranteed mycoplasma-free

Specificity: human & murine IL-24; human IL-20

Detection ranges:

• 3 - 100 ng/ml for human & murine IL-20
• 1 - 100 ng/ml for human & murine IL-24

Contents

Note: Each cell line is sold separately.

HEK-Blue™ IL-19/IL-20 cells

• 1 vial containing 3-7 x 10⁶ cells
• 1 ml Blasticidin (10 mg/ml)
• 1 ml Puromycin (10 mg/ml)
• 1 ml Zeocin® (100 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).

HEK-Blue™ IL-20 cells

• 1 vial containing 3-7 x 10⁶ cells
• 1 ml Blasticidin (10 mg/ml)
• 1 ml Puromycin (10 mg/ml)
• 1 ml Zeocin® (100 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 20 (IL-20) subfamily belongs to the IL-10 cytokine family and comprises IL-19, IL-20, IL-22, IL-24, and IL-26 [1].

IL-19, IL-20, IL-22, and IL-24 are closely-related cytokines mainly produced by monocytes and epithelial cells [1-2] and exerting their actions on a wide range of immune (i.e. monocytes and T cells) and non-immune cell types (e.g. epithelial cells and keratinocytes), especially in the barrier tissues  (e.g. gut, lung, skin) [2].

IL-20 signals through two receptors, which are shared with IL-19 and IL-24: they share different receptors

• IL-20 type I receptor is composed of the IL-20Rα (aka IL-20R1) and IL-20Rβ (aka IL-20R2) chains, and binds IL-19, IL-20, and IL-24.
• IL-20 type II receptor is composed of the IL-22Rα1 (aka IL-22R1) and IL-20Rβ chains and binds IL-20 and IL-24.

The signaling occurs through tyrosine kinases JAK1/2, Tyk2, and signal transducer and transcription activators (STAT). STAT3 plays a major role in the biological actions of IL-19, IL-20, IL-22, and IL-24. However, some functions have been also attributed and suggested for STAT1 and STAT5 [1]. 

IL-20 subfamily cytokines are upregulated in many human diseases, such as psoriasis, inflammatory bowel disease, and rheumatoid arthritis (RA). They promote downstream inflammatory responses, especially when synergizing with other cytokines, such as IL-17 and IFN-γ, and contribute to the pathogenesis of these diseases. On the other hand, these cytokines can enhance tissue regeneration and wound healing and thus may provide therapeutic potential in diseases associated with tissue damage [3]. 

References:

1. Caparrós E, Francés R., 2018. The Interleukin-20 Cytokine Family in Liver Disease. Front Immunol. 2018 May 28;9:1155.
2. Wei H, Li B, Sun A, Guo F., 2019 Interleukin-10 Family Cytokines Immunobiology and Structure. Adv Exp Med Biol. 2019;1172:79-96.
3. Ouyang W, O'Garra A., 2019. IL-10 Family Cytokines IL-10 and IL-22: from Basic Science to Clinical Translation. Immunity. 2019 Apr 16;50(4):871-891

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