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

作者：Zeuner Marie Theres; Vallance Thomas; Vaiyapuri Sakthivel; Cottrell Graeme S; Widera Darius^*

来源：Mediators of Inflammation, 2017, 2017: 6209865.

DOI：10.1155/2017/6209865

Aberrant activation of the transcription factor NF-kappa B, 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-kappa B-driven neuroinflammation has not been available. Thus, we developed a stable neural NF-kappa B-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-kappa B reporter construct containing GFP and firefly luciferase allowing an assessment of NF-kappa B activity via fluorescence microscopy, flow cytometry, and luminometry. We observed a robust activation of NF-kappa B after exposure of the reporter cell line to tumour necrosis factor alpha (TNFa) and amyloid-beta peptide [1-42] as well as to LPS derived from Salmonella minnesota and Escherichia coli. Finally, we demonstrate that the U251-NF-kappa B-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.

• 出版日期2017