NF-κB Signalling Pathway: Generation and Characterization of a Reporter Cell Line of Human Origin View Full Text


Ontology type: schema:Chapter     


Chapter Info

DATE

2012

AUTHORS

Inés Tiscornia , Pablo Espósito , Valentina Porro , Paola Hernández , Hugo Cerecetto , Mercedes González , Eliezer Barreiro , Mariela Bollati-Fogolín

ABSTRACT

The NF-κB is a transcription factor that plays a key role in regulating immunity. In response to signals, NF-κB activation occurs via phosphorylation of its inhibitor, which dissociates from the NF-κB dimer allowing the translocation to the nucleus where it induces gene expression (Karin 18:6867–6874, 1999). The anti-inflammatory effects of probiotics (Ma et al. 72:5308–5314, 2004; O’Hara et al. 118:202–215, 2006) and compounds (Majumdar et al. 168:2644–2651, 2002) can proceed through the modulation of NF-κB activation. Pathway-specific reporter cell systems appear as useful tools to screen and unravel the mode of action of compounds. Here, we report on the generation and characterization of a human reporter cell line that allows studying the modulation of NF-κB activation by different anti-inflammatory agents. Human intestinal epithelial cells were transfected with pNF-κB-hrGFP plasmid, stimulated with a pro-inflammatory cocktail and GFP positive cells were sorted. The responsiveness of the cells towards different concentrations of TNF-α was tested and the clone showing the best performance was selected. HT-29-NF-κB-hrGFP cells were incubated 18 h in the presence of TNF-α and potentially probiotic bacteria or synthetic compounds and the level of GFP and IL-8 was determined. Our results revealed that the HT-29-NF-κB-hrGFP reporter cell line represents a valuable tool for a reliable screening of probiotics and compounds with anti-inflammatory activities. More... »

PAGES

261-263

References to SciGraph publications

Book

TITLE

Proceedings of the 21st Annual Meeting of the European Society for Animal Cell Technology (ESACT), Dublin, Ireland, June 7-10, 2009

ISBN

978-94-007-0883-9
978-94-007-0884-6

Identifiers

URI

http://scigraph.springernature.com/pub.10.1007/978-94-007-0884-6_40

DOI

http://dx.doi.org/10.1007/978-94-007-0884-6_40

DIMENSIONS

https://app.dimensions.ai/details/publication/pub.1043524124


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