TSLP Reporter HEK 293 Cells

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Thymic Stromal Lymphopoietin Reporter Cells

Signaling pathway in HEK-Blue™ TSLP cells

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

TSLP is an interleukin-7 (IL-7)-like cytokine with multiple functions, including cell proliferation and survival. It is best known as a critical mediator of type 2 immune responses [1].

 More details

Cell line description

HEK-Blue™ TSLP cells were generated by stable transfection with the gene encoding for the TSLPR and IL-7Ra chains of the human TSLP receptor, STAT5, and a STAT5-inducible secreted embryonic alkaline phosphatase (SEAP) reporter. The binding of TSLP 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™ TSLP cells detect human but not murine TSLP (see figures). Of note, these cells also respond, to a weaker extent, to human IFN-γ (see figures). However, they do not respond to other STAT5-signaling cytokines: IL-2, IL-7, IL-9, and IL-15 (see figures).

Key features

• Fully functional TSLP signaling pathway
• Readily assessable STAT5-inducible SEAP reporter activity
• Strong response to human (h) TSLP
• No response to murine (m) TSLP
• Weak response to hIFN-γ
• No response to hIL-2, hIL-7, hIL-9, and hIL-15

Applications

• Detection and quantification of human and murine TSLP activity
• Screening of anti-TSLP and anti-TSLP receptor antibodies
• Screening of small molecule inhibitors of the TSLP pathway

Reference:

1. Ebina-Shibuya R. and Leonard WJ., 2023. Role of thymic stromal lymphopoietin in allergy and beyond. Nat Rev Immunol, 23(1):24-37.

Figures

Dose-response of HEK-Blue™ TSLP cells to recombinant TSLP. Cells were stimulated with increasing concentrations of recombinant human TSLP (hTSLP) and murine TSLP (mTSLP). After overnight incubation, the STAT5-induced 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™ TSLP cellular response using a neutralizing antibody against human TSLP.  An increasing concentration of a neutralizing anti-hTSLP antibody was incubated with recombinant human TSLP (3 ng/ml) for 2 hours before the addition of HEK-Blue™ TSLP cells. After overnight incubation, SEAP activity in the cell culture supernatant was assessed using QUANTI-Blue™. Data are shown as a percentage (%) of activity, mean ± SEM.

Response of HEK-Blue™ TSLP cells to a panel of cytokines. Cells were stimulated with various human and murine recombinant cytokines: 10 ng/ml of hTSLP, mTSLP, hIFN-γ, hTNF-α, hIL-1β, hIL-2, hIL-4, hIL-6, hIL-7, hIL-9, hIL-15, hIL-27 and 1000 U/ml hIFN-α. After overnight incubation, SEAP activity was assessed using QUANTI‑Blue™ The OD at 630 nm is shown as mean ± SEM.

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®

Specificity: Detects human TSLP

Detection range:

• Detection range for human TSLP: 0.3 - 100 ng/ml 

Quality Control:

• SEAP reporter activity in response to TSLP is validated using functional assays.
• The stability for 20 passages following thawing is confirmed.
• These cells are tested for mycoplasma contamination. 

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 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

Thymic stromal lymphopoietin (TSLP) is a cytokine produced by epithelial and stromal cells in barrier tissues as well as innate immune cells such as dendritic cells, basophils and mast cells. It has pleiotropic actions on a wide range of immune and non-immune cell types [1]. TSLP belongs to the common γ chain (γc) cytokine family. Its receptor comprises the IL-7Rα (CD127) and TSLPR (CRLF2), a γc-like subunit. It signals through tyrosine kinases of the Janus family (JAK1 and JAK2) and signal transducer and transcription activators (STATs), notably STAT5 [1]. TSLP is a pleiotropic cytokine best known as a critical mediator of type 2 immune responses and a promoter of Th2 cell-mediated diseases (e.g. asthma) [1,2]. Emerging evidence indicates that TSLP is also implicated in viral infections, cancer, chronic inflammation and fat metabolism [1].

1. Ebina-Shibuya R. and Leonard WJ., 2023. Role of thymic stromal lymphopoietin in allergy and beyond. Nat Rev Immunol, 23(1):24-37.
2. Schmitt P. et al., 2024. TL1A is an epithelial alarmin that cooperates with IL-33 for initiation of allergic airway inflammation. J Exp Med. 221(6) e20231236.

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