HEK-Blue-Lucia™ TLR Cells

HEK-Blue-Lucia™ TLR cells (formerly named HEK-Dual™ TLRs (NF/IL8) cell lines) are a collection of the human embryonic kidney 293 (HEK293)-derived cell lines designed to provide a rapid, sensitive, and reliable method to screen and validate TLR agonists or antagonists.

These cells express two inducible reporter genes for SEAP (secreted embryonic alkaline phosphatase) and Lucia luciferase. Stimulation with TLR agonists triggers the activation of the artificial NF-κB-inducible promoter and the subsequent production of SEAP. It also promotes the expression of Lucia luciferase, which is knocked in (KI) downstream of the endogenous (more physiological) IL-8 promoter. IL-8 (interleukin 8) is a chemokine produced in response to TLR agonists in an NF-κB/AP-1-dependent manner [1-2].

This enables the double readout study of the NF-κB/AP-1 pathway, by monitoring the activity of SEAP and Lucia luciferase using QUANTI-Blue™ Solution (SEAP detection reagent) or QUANTI-Luc™ 4 Lucia/Gaussia (luciferase detection reagent), respectively. Thus, you may choose the readout depending on your laboratory equipment utilizing a spectrophotometer for SEAP or a luminometer for Lucia luciferase detection.

These cells also feature a triple knockout (KO) of TLR3, TLR5, and TNFR. Therefore, the parental cell line HEK-Blue-Lucia™ Null cells allows for the independent study of one single TLR when used in combination with:

References:

1. Roebuck KA. 1999. Regulation of interleukin-8 gene expression. J Interferon Cytokine Res:429-38.
2. Ohta K, et al. 2014. Toll-like receptor (TLR) expression and TLR‑mediated interleukin-8 production by human submandibular gland epithelial cells. Mol Med Rep. (5):2377-82.