TL1A reporter HEK 293 cells

Notification:  Reference #hkb-tl1a-av can only be ordered together with reference #hkb-tl1a.

TL1A responsive NF-κB/AP1-SEAP reporter assay

Signaling pathway in HEK-Blue™ TL1A cells

HEK-Blue™ TL1A cells are designed to monitor human TL1A-induced NF-κB/AP1 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™ TL1A cells respond specifically to recombinant human (h) and mouse (m) TL1A. Their reliable and consistent performance makes them suitable for release assays of activatory and inhibitory molecules such as Tulisokibart, a monoclonal antibody that targets TL1A and prevents its binding to its receptor (see figures).

Key features

• Readily assessable NF-κB/AP1 reporter activity
• Convenient readout using QUANTI-Blue™ Solution
• High sensitivity to human (h)TL1A and mouse (m)TL1A activity
• Stability guaranteed for 20 passages

Applications

• Therapeutic development
• Drug screening
• Release assay

Tumor necrosis factor (TNF)-like 1A (TL1A) drives the production of pro-inflammatory cytokines and differentiation of T helper subsets. Abnormal TL1A expression is linked to enteric autoimmune diseases (e.g. Crohn’s disease) and extends to other diseases including 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™ TL1A cells to recombinant human TL1A. Cells were stimulated with increasing concentrations of recombinant human TL1A. After overnight incubation, the NF-kB/AP1-induced SEAP activity was determined using QUANTI-Blue™, a SEAP detection reagent. Data are shown as optical density (OD) at 650 nm (mean ± SEM).

 

Response of HEK-Blue™ TL1A cells to a panel of cytokines. Cells were stimulated with various human and murine recombinant cytokines: 10 ng/ml of hTL1A, mTL1A, hAPRIL, hBAFF, hCD40L, hRANKL, hTNF-α, hIFN-γ, hIL-1β, hIL-6, 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.

Dose-dependent inhibition of HEK-Blue™ TL1A cell response using Tulisokibart biosimilar. Increasing concentrations of Anti-hTL1A Tulisokibart biosimilar (0.1 ng/ml - 1 µg/ml) were incubated with recombinant human TL1A (3 ng/ml) for 1 h before the addition of HEK-Blue™ TL1A cells. After overnight incubation, SEAP activity in the cell culture supernatant was assessed using QUANTI-Blue™ Solution. Data are shown in percentage of activity (mean ± SEM).

Specifications

Cell type: Epithelial

Tissue origin: Human Embryonic Kidney

Target: TL1A

Specificity: Human, Mouse

Reporter gene: SEAP

Antibiotic resistance: Blasticidin, Zeocin®

Detection range: 300 pg/ml - 30 ng/ml (hTL1A and mTL1A)

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
• 1 ml of Blasticidin (10 mg/ml)
• 1 ml of Zeocin® (100 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)

Details

Tumor necrosis factor-like cytokine 1A (TL1A), also known as vascular endothelial growth inhibitor (VEGI) or TNFSF15, is part of the tumor necrosis factor (TNF) superfamily.

TL1A is mainly produced by endothelial cells, as well as activated dendritic cells and macrophages. It is synthesized as a membrane-bound trimeric molecule. Alternative splicing or cleavage by the tumor necrosis factor-alpha converting enzyme (TACE) results in soluble TL1A [1]. Both forms bind the homotrimeric transmembrane death receptor 3 (DR3), triggering TRADD/TRAF2/RIP signaling, and ultimately leading to NF-kB and MAPK activation. Subsequent gene expression in target cells drives the production of pro-inflammatory cytokines and differentiation of T helper subsets [1]. Alternatively, as DR3 is a dual signaling death receptor, TRADD also associates with FADD/RIP3/caspase 8 to induce apoptosis or necroptosis [1].

In resting T cells, DR3 is expressed as a soluble decoy receptor, DcR3, that protects the cells from apoptosis. Activated T cells express transmembrane DR3. Their activity can thus be modulated upon TL1A sensing, from proliferation and differentiation to response termination.

Two decades after its discovery, TL1A has become a hot therapeutic target. Abnormal TL1A expression is linked to multiple enteric autoimmune diseases (e.g. Crohn’s disease, ulcerative colitis) and extends to other diseases, including rheumatoid arthritis, psoriasis, and allergic airway inflammation [1, 2]. Genome-wide association studies have also linked TL1A polymorphisms with disease susceptibility, conferring a biomarker relevance on TL1A [1].

Anti-TNFa monoclonal antibodies (mAbs) such as Adalimumab are among industry’s top-selling drugs to treat autoimmune diseases [3]. However, many patients either do not respond to their treatment or suffer from side-effects that are inherent to long-lasting TNF blockage [3]. Targeting TL1A could break the efficacy ceiling that we see with the best drugs out there. Pharmaceutical biotechs now have three neutralizing mAbs in clinical development. Inhibition of TL1A signaling using anti-TL1A Tulisokibart (MK-7240, from Merck), RVT-3101 (from Roche), or Duvakitub (TEV-48574, from Sanofi & Teva Pharmaceuticals) has shown clinical evidence of “best-in-class” potential for treating inflammation and scarring in inflammatory bowel disease (IBD), ulcerative colitis, and Crohn’s disease [4]. The TL1A /DR3 axis could also be targeted to enhance or restore immune responses, such as in exhausted T cells in a tumor environment. Further research should guarantee the clinical usage of DR3 agonists, or inhibitors of the soluble decoy receptor DcR3, either alone or in combination with immune checkpoint blockade.

​References:

1. Xu, W.-D, et al., 2021. Role of TL1A in Inflammatory Autoimmune Diseases: A Comprehensive Review. Frontiers in Immunology, 2022. 13:891328.
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). doi: 10.1084/jem.20231236.
3. Steeland, S., C. et al., 2018. A New Venue of TNF Targeting. International Journal of Molecular Sciences. 19(5):1442.
4. Neimark, J., 2024. TL1A inhibitors could usher in new era of IBD treatment. www.biospace.com. 

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