Development and Characterisation of a Novel NF-κB Reporter Cell Line for Investigation of Neuroinflammation
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 an...
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| Main Authors: | , , , , |
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| Format: | Article |
| Language: | English |
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Wiley
2017-01-01
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| Series: | Mediators of Inflammation |
| Online Access: | http://dx.doi.org/10.1155/2017/6209865 |
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| _version_ | 1832556643227795456 |
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| author | Marie-Theres Zeuner Thomas Vallance Sakthivel Vaiyapuri Graeme S. Cottrell Darius Widera |
| author_facet | Marie-Theres Zeuner Thomas Vallance Sakthivel Vaiyapuri Graeme S. Cottrell Darius Widera |
| author_sort | Marie-Theres Zeuner |
| collection | DOAJ |
| description | 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. |
| format | Article |
| id | doaj-art-e3928681c3fc4edb92c6d30641aab78a |
| institution | Kabale University |
| issn | 0962-9351 1466-1861 |
| language | English |
| publishDate | 2017-01-01 |
| publisher | Wiley |
| record_format | Article |
| series | Mediators of Inflammation |
| spelling | doaj-art-e3928681c3fc4edb92c6d30641aab78a2025-02-03T05:44:55ZengWileyMediators of Inflammation0962-93511466-18612017-01-01201710.1155/2017/62098656209865Development and Characterisation of a Novel NF-κB Reporter Cell Line for Investigation of NeuroinflammationMarie-Theres Zeuner0Thomas Vallance1Sakthivel Vaiyapuri2Graeme S. Cottrell3Darius Widera4Stem Cell Biology and Regenerative Medicine, School of Pharmacy, University of Reading, Reading RG6 6AP, UKStem Cell Biology and Regenerative Medicine, School of Pharmacy, University of Reading, Reading RG6 6AP, UKSchool of Pharmacy, University of Reading, Reading RG6 6AP, UKCellular and Molecular Neuroscience, School of Pharmacy, University of Reading, Reading RG6 6AP, UKStem Cell Biology and Regenerative Medicine, School of Pharmacy, University of Reading, Reading RG6 6AP, UKAberrant 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.http://dx.doi.org/10.1155/2017/6209865 |
| spellingShingle | Marie-Theres Zeuner Thomas Vallance Sakthivel Vaiyapuri Graeme S. Cottrell Darius Widera Development and Characterisation of a Novel NF-κB Reporter Cell Line for Investigation of Neuroinflammation Mediators of Inflammation |
| title | Development and Characterisation of a Novel NF-κB Reporter Cell Line for Investigation of Neuroinflammation |
| title_full | Development and Characterisation of a Novel NF-κB Reporter Cell Line for Investigation of Neuroinflammation |
| title_fullStr | Development and Characterisation of a Novel NF-κB Reporter Cell Line for Investigation of Neuroinflammation |
| title_full_unstemmed | Development and Characterisation of a Novel NF-κB Reporter Cell Line for Investigation of Neuroinflammation |
| title_short | Development and Characterisation of a Novel NF-κB Reporter Cell Line for Investigation of Neuroinflammation |
| title_sort | development and characterisation of a novel nf κb reporter cell line for investigation of neuroinflammation |
| url | http://dx.doi.org/10.1155/2017/6209865 |
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