Notification: Reference #hkd-htlr2ni-av can only be ordered together with reference #hkd-htlr2ni.

NF-κB–SEAP and IL-8–Lucia reporter HEK293 cells expressing human TLR2

Signaling pathways in HEK-Blue-Lucia™ hTLR2 cells

InvivoGen offers a human embryonic kidney 293 (HEK293)-derived cell line, specifically designed to assess the distinct role of the human Toll-like receptor 2 (hTLR2):

— HEK-Blue-Lucia™ hTLR2 cells*

These cells were generated from the HEK-Blue-Lucia™ Null cell line harboring two inducible reporter genes. This feature allows the double readout of the NF-κB/AP-1 pathway, by monitoring the SEAP (secreted embryonic alkaline phosphatase) or Lucia luciferase activities. HEK-Blue-Lucia™ hTLR2 cells also stably express the hTLR2 and CD14 genes. Due to a triple knockout (KO) of TLR3, TLR5, and TNFR, this cell line allows for the independent study of TLR2.

Stimulation of HEK-Blue-Lucia™ hTLR2 cells with TLR2 agonists (e.g. Pam3CSK4) 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 (see figures).

IL-8 (interleukin 8) is a chemokine produced in response to TLR agonists in an NF-κB/AP-1-dependent manner [1-2]. This feature 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.

TLR2 is an important pattern recognition receptor (PRR) detecting a large spectrum of microbial pathogen-associated molecular patterns (PAMPs) from Gram-positive and Gram-negative bacteria as well as fungi, parasites, and viruses [3].

Read our review on TLR2

Key features:

Stable overexpression of hTLR2 and CD14
Verified KO for the TLR3, TLR5, and TNFR genes
Functionally validated using a selection of PRR ligands and cytokines
Readily assessable NF-κB activation by assessing the SEAP and/or Lucia luciferase activities

Applications:

Defining the role of TLR2-dependent NF-κB signaling pathway
Screening for novel TLR2 agonists and inhibitors
Choice of readout depending on the laboratory equipment (spectrophotometer for SEAP or luminometer for Lucia luciferase detection).

*Note: This cell line has been renamed. It was formerly known as "HEK-Dual™ hTLR2 (NF/IL8)". The cat. code (hkd-htlr2ni) remains unchanged.

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.
3. Oliveira-Nascimento, L. et al. 2012. The Role of TLR2 in Infection and Immunity. Front Immunol 3, 79.

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