IL-7 Reporter HEK 293 Cells

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Interleukin-7 Reporter Cells

Signaling pathway in HEK-Blue™ IL-7 cells

HEK-Blue™ IL-7 cells were engineered from the human embryonic kidney HEK 293 cell line to detect bioactive interleukin-7 (IL-7) by monitoring the activation of the JAK/STAT5 pathway. They can also be used for screening antibodies or small molecule inhibitors targeting the IL-7 pathway.

IL-7 is a secreted cytokine that plays an essential role in B cell and T cell development and function [1-4]. 

 More details

Cell line description

HEK-Blue™ IL-7 cells were generated by stable transfection with the genes encoding the human IL-7 receptor (IL-7Rα and IL-2Rγ chains), human JAK3, human STAT5b, and a STAT5-inducible secreted embryonic alkaline phosphatase (SEAP) reporter. The binding of IL-7 to its receptor triggers a signaling cascade leading to the activation of STAT5 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-7 cells detect human and murine IL-7 (see figures). Of note, HEK‑Blue™ IL-7 cells also exhibit a moderate response to human IFN-γ but do not respond to human IFN-α (see figures).

Key features

• Fully functional IL-7 signaling pathway
• Readily assessable STAT5-inducible SEAP reporter activity
• Strong response to human (h) and mouse (m) IL-7

Applications

• Detection and quantification of hIL-7 and mIL-7 activity
• Screening of anti-IL-7 and anti-IL-7 receptor antibodies
• Screening of small molecule inhibitors of the IL-7 pathway

References:

1. Lin J. et al., 2017. The role of IL-7 in Immunity and Cancer. Anticancer Res. 37(3):963-7.
2. Ribeir D. et al., 2018. STAT5 is essential for IL-7-mediated viability, growth, and proliferation of T-cell acute lymphoblastic leukemia cells. Blood Adv. 2(17):2199-213.
3. Mackall C.L. et al., 2011. Harnessing the biology of IL-7 for therapeutic application. Nat Rev Immunol. 11(5):330-42.
4. Barata J.T. et al., 2019. Flip the coin: IL-7 and IL-7R in health and disease. Nat Immunol. 20(12):1584-93.

Figures

Validation of the expression of human IL-7Rα by HEK-Blue™ IL-7 cells.
5 x 10⁵ cells were incubated with either an APC-conjugated isotype control (blue) or an APC-conjugated Anti-IL-7Rα mAb (red) for 30 minutes. The binding affinity was then measured using flow cytometry.

Dose-response of HEK-Blue™ IL-7 cells to recombinant IL-7.
Cells were stimulated with increasing concentrations of recombinant human IL-7 (hIL-7) and murine IL-7 (mIL-7). After overnight incubation, the STAT5 response was determined using QUANTI‑Blue™ Solution, a SEAP detection reagent. The optical density (OD) at 630 nm is shown as mean ± SEM.

Dose-dependent inhibition of HEK‑Blue™ IL-7 cellular response using a neutralizing antibody against IL-2Rγ, a subunit of IL‑7R.
The antibody was incubated with HEK-Blue™ IL-7 cells for 2 hours prior to the addition of hIL-7 (0.3 ng/ml). After overnight incubation, SEAP activity in the cell culture supernatant was assessed using QUANTI-Blue™ Solution. Data are shown as a percentage (%) of activity.

Response of HEK-Blue™ IL-7 cells to a panel of cytokines.
Cells were stimulated with various human and murine recombinant cytokines: 1 ng/ml of hIL-7 or mIL-7, and 100 ng/ml hIL-2, hIL-4, hIL‑6, hIL-9, mIL-9, hIL‑15, hIL-21, or hIL-27, and 1000 U/ml hIFN-α, and 10 ng/ml hIFN-γ. After overnight incubation, SEAP activity was assessed using QUANTI‑Blue™ Solution. The optical density (OD) at 630 nm is shown as 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-7 and mouse IL-7

Detection range:

• Detection range for human IL-7: 100 pg/ml - 100 ng/ml
• Detection range for murine IL-7: 100 pg/ml - 100 ng/ml

Quality Control:

• SEAP reporter activity in response to IL-7 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 7 (IL-7) is a secreted cytokine that plays an essential role in B cell and T cell development and function [1]. IL-7 signals through the heterodimeric cell surface IL-7 receptor (IL-7R) consisting of IL-7Rα (also called CD127) and IL-2Rγ (also called the common γ-chain or CD132).
The binding of IL-7 to its receptor triggers three main signaling pathways: JAK/STAT, PI3K, and MAPK/ERK [1‑3]. Of note, IL-7R-mediated signaling triggers proliferative and anti‑apoptotic signals mainly by activating the JAK/STAT pathway [1-3]. IL-7/IL-7R signaling, which regulates lymphocyte growth and survival, has been implicated in the development of malignancies and autoimmune diseases [1-4].

1. Lin J. et al., 2017. The role of IL-7 in Immunity and Cancer. Anticancer Res. 37(3):963-7.
2. Ribeir D. et al., 2018. STAT5 is essential for IL-7-mediated viability, growth, and proliferation of T-cell acute lymphoblastic leukemia cells. Blood Adv. 2(17):2199-213.
3. Mackall C.L. et al., 2011. Harnessing the biology of IL-7 for therapeutic application. Nat Rev Immunol. 11(5):330-42.
4. Barata J.T. et al., 2019. Flip the coin: IL-7 and IL-7R in health and disease. Nat Immunol. 20(12):1584-93.

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