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Mediators of Inflammation
Volume 2017, Article ID 6209865, 10 pages
https://doi.org/10.1155/2017/6209865
Research Article

Development and Characterisation of a Novel NF-κB Reporter Cell Line for Investigation of Neuroinflammation

1Stem Cell Biology and Regenerative Medicine, School of Pharmacy, University of Reading, Reading RG6 6AP, UK
2School of Pharmacy, University of Reading, Reading RG6 6AP, UK
3Cellular and Molecular Neuroscience, School of Pharmacy, University of Reading, Reading RG6 6AP, UK

Correspondence should be addressed to Darius Widera; ku.ca.gnidaer@arediw.d

Received 22 March 2017; Accepted 19 June 2017; Published 16 July 2017

Academic Editor: Soh Yamazaki

Copyright © 2017 Marie-Theres Zeuner et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

Abstract

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 been available. Thus, we developed a stable neural NF-κB-reporter cell line to assess the potency of proinflammatory 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 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 biologics in neural cells.