IL-33 Reporter HEK 293 Cells

IL-33 Reporter Cells

IL-33 signaling in HEK-Blue™ IL-33 Cells

HEK-Blue™ IL-33 cells are designed to detect bioactive interleukin-33 (IL-33) by monitoring the activation of the NF-κB and AP-1 pathways. IL-33 is a pro-inflammatory cytokine that shares structural and functional characteristics with the IL-1 cytokine family. It binds and signals through the IL-1RL1/ IL-1R accessory protein (IL-1RAcP) receptor activating NF-κB and MAP kinases.

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Cell line description:

HEK-Blue™ IL-33 cells were generated by stable transfection of human embryonic kidney HEK293-derived cells with the human IL1RL1 gene. In addition, the TNF-α and the IL-1β responses have been blocked.

Therefore, HEK-Blue™ IL-33 cells respond specifically to IL-33. These cells express an NF-κB/AP-1- inducible SEAP reporter gene.

The binding of human IL-33 to the heterodimeric  IL-1RL1/ IL-1RAcP  on the surface of these cells triggers a signaling cascade leading to the activation of NF-κB and the subsequent production of SEAP.

Levels of SEAP in the supernatant can be easily determined with QUANTI-Blue™ Solution.

Features of HEK-Blue™ IL-33 cells:

• Fully functional IL-33 signaling pathway
• Do not respond to human TNF-α
• Do not respond to human IL-1β
• Readily assessable SEAP reporter activity
• Functionally tested and guaranteed mycoplasma-free

Applications of HEK-Blue™ IL-33 cells:

• Detection of human IL-33
• Screening of anti-IL-33 antibodies

Figures

Dose-response of HEK-Blue™ IL‑33 cells to recombinant hIL‑33.
Cells were stimulated with increasing concentrations of recombinant IL-33. After overnight incubation, the NF-κB/AP-1 response was determined using QUANTI-Blue™ Solution, a SEAP detection reagent, and reading the optical density (OD) at 630 nm. Data are shown as mean ± SEM.

Response of HEK‑Blue™ IL‑33 cells to a panel of cytokines.
Cells were stimulated with various human and murine recombinant cytokines: 1 ng/ml of hIL‑33, 10 ng/ml of hIL‑1β, mIL-1β, hIL-2, hIL-4, hIL-17, hIL-18, hIFN-γ, hTNF-α, mTNF-α, or 102 U/ml hIFN-α2a or hIFN-β. After overnight incubation, SEAP activity was assessed using QUANTI-Blue™ Solution. The OD readings at 630 nm are shown as mean ± SEM.

Dose-dependent inhibition of HEK-Blue™ IL‑33 cellular response using a neutralizing antibody against hIL‑33.
The anti‑hIL33 antibody was incubated with the cells for 30 minutes prior to the addition of hIL-33 (1 ng/ml). After overnight incubation, SEAP activity in the cell culture supernatant was assessed using QUANTI-Blue™ Solution. Data (shown as mean ± SEM) represent the percentage (%) of maximal reporter activity without the anti-hIL33 antibody.

Specifications

Antibiotic resistance: blasticidin, hygromycin B, 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: Detects human IL-33

Detection range: 0.5 - 100 ng/ml

Contents

• 1 vial containing 3-7 x 10⁶ cells
• 2 x 1 ml of HEK-Blue™ Selection (250x concentrate)
• 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)

Details

Interleukin-33 (IL-33; also known as IL-1F11) is a member of the IL-1 family, a group of cytokines that play important roles in host defense, immune regulation, and inflammation [1]. IL-33 mediates its biological effects through  IL1RL1  (also known as ST2), a receptor expressed on Th2 and mast cells. IL-33 and  IL1RL1 form a complex with IL-1R accessory protein (IL-1RAcP), a signaling receptor subunit that is also a member of the IL-1R complex. IL-33 signaling leads to the activation of NF-κB and MAP kinases.

IL-33 can function both as a traditional cytokine and as a nuclear factor regulating gene transcription. Following pro-inflammatory stimulation, Il-33 can induce Th2-biased immune responses, such as the production of IL-4, IL-5 and IL-13 [2]. In addition, as IL-33 is constitutively expressed in endothelial and epithelial cells, it can act as an endogenous danger signal, or damage-associated molecular pattern (DAMP; also called alarmins), in response to tissue damage [3, 4].

1. Arend W. et al., 2008. IL-1, IL-18, and IL-33 families of cytokines. Immunol Rev. 223:20-38.
2. Schiering C. et al., 2014. The alarmin IL-33 promotes regulatory T-cell function in the intestine. Nature. 513(7519):564-8.
3. Cayrol C. & Girard JP., 2014. IL-33: an alarmin cytokine with crucial roles in innate immunity, inflammation and allergy. Curr Opin Immunol. 31-7.
4. Oboki K. et al., 2011. IL-33 and airway inflammation. Allergy Asthma Immunol Res. 3(2): 81–88 

FAQ

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