In:
Mediators of Inflammation, Hindawi Limited, Vol. 2017 ( 2017), p. 1-10
Abstract:
Aberrant activation of the transcription factor NF- κ 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- κ B-driven neuroinflammation has not been available. Thus, we developed a stable neural NF- κ 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- κ B reporter construct containing GFP and firefly luciferase allowing an assessment of NF- κ B activity via fluorescence microscopy, flow cytometry, and luminometry. We observed a robust activation of NF- κ B after exposure of the reporter cell line to tumour necrosis factor alpha (TNF α ) 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- κ 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.
Type of Medium:
Online Resource
ISSN:
0962-9351
,
1466-1861
DOI:
10.1155/2017/6209865
Language:
English
Publisher:
Hindawi Limited
Publication Date:
2017
detail.hit.zdb_id:
2008065-7
