TSLP Reporter HEK 293 Cells

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TSLP responsive STAT5-SEAP reporter cells

Signaling pathway in HEK-Blue™ TSLP cells

HEK-Blue™ TSLP cells are designed to monitor human TSLP-induced STAT5 stimulation or inhibition. This colorimetric bioassay can be used to screen activatory or inhibitory molecules, such as engineered cytokines and neutralizing antibodies, respectively.

HEK-Blue™ TSLP cells respond specifically to recombinant human TSLP. Their reliable and consistent performance makes them suitable for release assays of activatory and inhibitory molecules such as Tezepelumab, a monoclonal antibody that targets TSLP and prevents its binding to its receptor (see figures).

Key features

• Readily assessable STAT5-SEAP reporter activity
• Convenient readout using QUANTI-Blue™ Solution
• High sensitivity to human TSLP activity
• No response to mouse TSLP
• Stability guaranteed for 20 passages

Applications

• Therapeutic development
• Drug screening
• Release assay

Thymic stromal lymphopoietin (TSLP) is a critical mediator of type 2 (T2) immune responses, acting at the early stages of inflammation. Abnormal TSLP expression is linked to immune-mediated inflammation diseases, notably asthma and allergic airway inflammation.

 More details

InvivoGen’s products are for research use only, and not for clinical or veterinary use.

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

Cell type: Epithelial

Tissue origin: Human Embryonic Kidney

Target: TSLP

Specificity: Human

Reporter gene: SEAP

Antibiotic resistance: Blasticidin, Hygromycin B, Zeocin®

Detection range: 0.3 ng/ml - 100 ng/ml (hTSLP)

Growth medium: Complete DMEM (see TDS)

Growth properties: Adherent

Mycoplasma-free: Verified using Plasmotest™

Quality control: Each lot is functionally tested and validated.

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)

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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