IL-17 Reporter HEK 293 Cells

Cytokine offer: For each cytokine reporter cell line purchased, get a free vial of the matching cytokine. View all InvivoGen's Cytokine Bioassays.

IL-17 Reporter Cells

Signaling pathway in HEK-Blue™ IL-17 cells

HEK-Blue™ IL-17 cells are designed to monitor human IL-17-induced NF-κB/AP-1 stimulation or inhibition through SEAP detection. This colorimetric bioassay can be used for screening activatory molecules, such as engineered cytokines, or inhibitory molecules, such as neutralizing antibodies.

HEK-Blue™ IL-17 cells respond specifically to recombinant human IL-17A, human IL-17F, as well as human and mouse IL-17E. The reliable and consistent performance of HEK-Blue™ IL-17 cells makes them suitable for release assays of therapeutic molecules that inhibit IL-17 signaling, such as Secukinumab, a monoclonal antibody targeting the IL-17A (see figures).

Key features

• Readily assessable NF-κB/AP-1-SEAP reporter activity
• Convenient readout using  QUANTI-Blue™ Solution
• Strong response to human (h)IL-17A, hIL-17F, & mouse (m)/hIL-17E
• Low to no response to mIL-17A, h/mIL-17C, & mIL-17F
• Stability guaranteed for 20 passages

Applications

• Therapeutic development
• Drug screening
• Release assay

The interleukin-17 (IL-17) family comprises six members (IL-17A – 17F), which have various biological functions, including driving an inflammatory cascade during infections and autoimmune diseases. IL-17A is a key therapeutic target for the treatment of moderate-to-severe plaque psoriasis, psoriatic arthritis, and ankylosing spondylitis.

 More details

Figures

Dose-response of HEK-Blue™ IL-17 cells to recombinant human IL-17A. HEK-Blue™ IL-17 cells were stimulated with increasing concentrations of recombinant IL-17A. After overnight incubation, the NF-kB/AP-1-induced SEAP activity was determined using QUANTI-Blue™ Solution, a SEAP detection reagent. Data are shown as optical density (OD) at 650 nm (mean ± SEM).

Dose-dependent inhibition of HEK-Blue™ IL-17 cell response using Secukinumab biosimilar. Increasing concentrations of Anti-hIL-17-hIgG1 (0.1 ng/ml - 10 µg/ml) were incubated with recombinant human IL-17A (1 ng/ml) for 1 hour prior to the addition of HEK-Blue IL-17 cells. After overnight incubation, SEAP activity in the cell culture supernatant was assessed using QUANTI-Blue™ Solution. Data are shown as the percentage of activity (mean ± SEM).

Cytokine response profile of HEK-Blue™ IL-17 cells. Cells were stimulated with various human and mouse recombinant cytokines: 10 ng/ml of hIL‑17A, mIL‑17A, hIL‑17C, hIL‑17E (hIL-25), and hIL‑17F. After overnight incubation, SEAP activity was assessed using QUANTI‑Blue™ Solution. Data are shown as optical density (OD) at 650 nm (mean ± SEM).

Specifications

Cell type: Epithelial

Tissue origin: Human Embryonic Kidney

Target: IL-17A, IL-17F,  IL-17E (IL-25)

Specificity: Human, mouse

Reporter gene: SEAP

Antibiotic resistance: Blasticidin, Hygromycin, Zeocin®

Detection ranges: 

• human IL-17A: 1- 100 ng/ml
• human & mouse IL-17E (IL-25): 1- 100 ng/ml
• human IL-17F: 3 - 100 ng/ml

Growth medium: Complete DMEM (see TDS)

Growth properties: Adherent

Mycoplasma-free: Verified using Plasmotest™

Quality control: Each lot is functionally tested and validated.

Contents

HEK-Blue™ IL-17 Cells (hkb-il17)

• 1 vial containing 3-7 x 10⁶ cells
• 2 x 1 ml HEK-Blue Selection (250X concentrate)
• 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-17 vial (hkb-il17-av)

• 1 vial containing 3-7 x 10⁶ cells

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

Notification:  Reference #hkb-il17-av can only be ordered together with reference #hkb-il17.

Details

Cell line description

HEK-Blue™ IL-17 cells were generated by stable transfection with the genes encoding for the human IL-17 receptor (IL-17RA and IL-17RC chains), the ACT1 adaptor molecule, as well as an NF-κB- and AP-1-inducible secreted embryonic alkaline phosphatase (SEAP) reporter. The binding of IL-17 to its receptor triggers a signaling cascade leading to the activation of NF-κB/AP-1 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-17 cells detect human (h) IL-17A, hIL-17E, hIL-17F, and murine (m) IL-17E. They display little to no response to mIL‑17A and mIL-17F. They do not respond to hIL-17C, mIL-17C. 

Interleukin-17 background

Interleukin 17 (IL-17) is a family of six closely related cytokines (IL-17A to IL-17F) which have both pro-inflammatory and anti-inflammatory activities. IL-17A and IL-17F, which can form a heterodimer, play an important role in Th17 immunity and are implicated in tumorigenesis and autoimmune diseases, whereas IL-17E (also known as IL-25) appears to promote Th2 immunity [1,2].

IL-17 cytokines exert their biological activities by binding to heterodimeric receptors containing the ubiquitous IL-17RA chain and a second IL-17R(C, B, or E) chain. IL-17A and IL-17F bind to the IL-17RA/IL-17RC receptor, IL-17C binds to the IL-17RA/IL-17RE receptor, and IL-17E binds to the IL-17RA/IL-17RB receptor [1, 2]. The activated heterodimeric receptor recruits the Act1 adaptor and induces the TNF receptor-associated factor 6 (TRAF6) ubiquitylation. This triggers a signaling cascade that results in NF-κB and AP-1 activation [1].

Antibodies targeting IL-17A, namely Secukinumab and Ixekizumab, were approved in 2016 for the treatment of moderate-to-severe plaque psoriasis, ankylosing spondylitis (AS), and psoriatic arthritis (PsA). Surprisingly, trials of Secukinumab and Brodalumab, an anti-IL-17RA in Crohn’s disease, were terminated early due to worsening of the disease in the treatment group [3-4]. These findings agree with IL-17's supportive role in skin wound healing [3].

1. Monin L. & Gaffen S.L., 2018. Interleukin 17 family cytokines: signaling mechanisms, biological activities, and therapeutic implications. Cold Spring Harb Perspect Biol. 10(4).
2. Pappu R. et al., 2011. The interleukin-17 cytokine family: critical players in host defence and inflammatory diseases. Immunology. 134: 8-16.
3. Huangfu L, Li R, Huang Y, Wang S, 2023. The IL-17 family in diseases: from bench to bedside. Signal Transduct Target Ther.11;8(1):402.
4. Amatya N, Garg AV, Gaffen SL, 2017. IL-17 Signaling: The Yin and the Yang. Trends Immunol. 38(5):310-322.

FAQ Cell Lines

Any questions about our cell lines ? Visit our frequently asked questions page