Development of an in Vitro Potency Assay for Anti-anthrax Lethal Toxin Neutralizing Antibodies

Lethal toxin (LT) of Bacillus anthracis reduces the production of a number of inflammatory mediators, including transcription factors, chemokines and cytokines in various human cell lines, leading to down-regulation of the host inflammatory response. Previously we showed that the reduction of interl...

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Main Authors:	Whiting, Gail, Baker, Michael, Rijpkema, Sjoerd
Format:	Online
Language:	English
Published:	MDPI 2012
Online Access:	https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3277096/

id	pubmed-3277096
recordtype	oai_dc
spelling	pubmed-32770962012-02-15 Development of an in Vitro Potency Assay for Anti-anthrax Lethal Toxin Neutralizing Antibodies Whiting, Gail Baker, Michael Rijpkema, Sjoerd Article Lethal toxin (LT) of Bacillus anthracis reduces the production of a number of inflammatory mediators, including transcription factors, chemokines and cytokines in various human cell lines, leading to down-regulation of the host inflammatory response. Previously we showed that the reduction of interleukin-8 (IL-8) is a sensitive marker of LT-mediated intoxication in human neutrophil-like NB-4 cells and that IL-8 levels are restored to normality when therapeutic monoclonal antibodies (mAb) with toxin-neutralising (TN) activity are added. We used this information to develop cell-based assays that examine the effects of TN therapeutic mAbs designed to treat LT intoxication and here we extend these findings. We present an in vitro assay based on human endothelial cell line HUVEC jr2, which measures the TN activity of therapeutic anti-LT mAbs using IL-8 as a marker for intoxication. HUVEC jr2 cells have the advantage over NB-4 cells that they are adherent, do not require a differentiation step and can be used in a microtitre plate format and therefore can facilitate high throughput analysis. This human cell-based assay provides a valid alternative to the mouse macrophage assay as it is a more biologically relevant model of the effects of toxin-neutralising antibodies in human infection. MDPI 2012-01-19 /pmc/articles/PMC3277096/ /pubmed/22347621 http://dx.doi.org/10.3390/toxins4010028 Text en © 2012 by the authors; licensee MDPI, Basel, Switzerland. http://creativecommons.org/licenses/by/3.0/ This article is an open-access article distributed under the terms and conditions of the Creative Commons Attribution license (http://creativecommons.org/licenses/by/3.0/).
repository_type	Open Access Journal
institution_category	Foreign Institution
institution	US National Center for Biotechnology Information
building	NCBI PubMed
collection	Online Access
language	English
format	Online
author	Whiting, Gail Baker, Michael Rijpkema, Sjoerd
spellingShingle	Whiting, Gail Baker, Michael Rijpkema, Sjoerd Development of an in Vitro Potency Assay for Anti-anthrax Lethal Toxin Neutralizing Antibodies
author_facet	Whiting, Gail Baker, Michael Rijpkema, Sjoerd
author_sort	Whiting, Gail
title	Development of an in Vitro Potency Assay for Anti-anthrax Lethal Toxin Neutralizing Antibodies
title_short	Development of an in Vitro Potency Assay for Anti-anthrax Lethal Toxin Neutralizing Antibodies
title_full	Development of an in Vitro Potency Assay for Anti-anthrax Lethal Toxin Neutralizing Antibodies
title_fullStr	Development of an in Vitro Potency Assay for Anti-anthrax Lethal Toxin Neutralizing Antibodies
title_full_unstemmed	Development of an in Vitro Potency Assay for Anti-anthrax Lethal Toxin Neutralizing Antibodies
title_sort	development of an in vitro potency assay for anti-anthrax lethal toxin neutralizing antibodies
description	Lethal toxin (LT) of Bacillus anthracis reduces the production of a number of inflammatory mediators, including transcription factors, chemokines and cytokines in various human cell lines, leading to down-regulation of the host inflammatory response. Previously we showed that the reduction of interleukin-8 (IL-8) is a sensitive marker of LT-mediated intoxication in human neutrophil-like NB-4 cells and that IL-8 levels are restored to normality when therapeutic monoclonal antibodies (mAb) with toxin-neutralising (TN) activity are added. We used this information to develop cell-based assays that examine the effects of TN therapeutic mAbs designed to treat LT intoxication and here we extend these findings. We present an in vitro assay based on human endothelial cell line HUVEC jr2, which measures the TN activity of therapeutic anti-LT mAbs using IL-8 as a marker for intoxication. HUVEC jr2 cells have the advantage over NB-4 cells that they are adherent, do not require a differentiation step and can be used in a microtitre plate format and therefore can facilitate high throughput analysis. This human cell-based assay provides a valid alternative to the mouse macrophage assay as it is a more biologically relevant model of the effects of toxin-neutralising antibodies in human infection.
publisher	MDPI
publishDate	2012
url	https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3277096/
_version_	1611505452329205760

Development of an in Vitro Potency Assay for Anti-anthrax Lethal Toxin Neutralizing Antibodies

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