Development and characterisation of a novel NF-κB reporter cell line for investigation of neuroinflammation

Zeuner, M.-T., Vallance, T., Vaiyapuri, S. ORCID: https://orcid.org/0000-0002-6006-6517, Cottrell, G. S. ORCID: https://orcid.org/0000-0001-9098-7627 and Widera, D. ORCID: https://orcid.org/0000-0003-1686-130X (2017) Development and characterisation of a novel NF-κB reporter cell line for investigation of neuroinflammation. Mediators of inflammation, 2017. 6209865. ISSN 0962-9351 doi: 10.1155/2017/6209865

Abstract/Summary

Aberrant activation of the transcription factor NF-κB, as well as uncontrolled inflammation has been linked to autoimmune diseases, development and progression of cancer and neurological disorders like Alzheimer’s disease. Reporter cell lines are a valuable state-of-the art tool for comparative analysis of in vitro drug screening. However, a reporter cell line for the investigation of NF-κB-driven neuroinflammation has not yet been available. Thus, we developed a stable neural NF-κB-reporter cell line to assess the potency of pro-inflammatory molecules and peptides, as well as anti-inflammatory pharmaceuticals. We used lentivirus to transduce the glioma cell line U251-MG with a tandem NF-κB reporter construct containing GFP and firefly luciferase allowing an assessment of NF-κB activity via fluorescence microscopy, flow cytometry and luminometry. We observed a robust activation of NF-κB after exposure of the reporter cell line to Tumour 2 necrosis factor alpha (TNFα) and amyloid-β peptide [1-42] as well as to LPS derived from Salmonella minnesota and Escherichia coli. Finally, we demonstrate that the U251-NF-κB-GFP-Luc reporter cells can be used for assessing the anti-inflammatory potential of pharmaceutical compounds using Bay11-7082 and IMD0354. In summary, our newly generated cell line is a robust and cost-efficient tool to study pro- and anti-inflammatory potential of drugs and biologicals in neural cells.

Item Type	Article
URI	https://reading-clone.eprints-hosting.org/id/eprint/70869
Identification Number/DOI	10.1155/2017/6209865
Refereed	Yes
Divisions	Life Sciences > School of Chemistry, Food and Pharmacy > School of Pharmacy > Division of Pharmacology
Publisher	Hindawi Publishing Corporation
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