Polymyxin B as inhibitor of LPS contamination of <it>Schistosoma mansoni </it>recombinant proteins in human cytokine analysis

<p>Abstract</p> <p>Background</p> <p>Recombinant proteins expressed in <it>Escherichia coli </it>vectors are generally contaminated with endotoxin. In this study, we evaluated the ability of Polymyxin B to neutralize the effect of LPS present as contaminant...

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Bibliographic Details
Main Authors: Pacífico Lucila G, Góes Alfredo M, Araujo Maria, Cardoso Luciana S, Oliveira Ricardo R, Oliveira Sergio C
Format: Article
Language:English
Published: BMC 2007-01-01
Series:Microbial Cell Factories
Online Access:http://www.microbialcellfactories.com/content/6/1/1
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Summary:<p>Abstract</p> <p>Background</p> <p>Recombinant proteins expressed in <it>Escherichia coli </it>vectors are generally contaminated with endotoxin. In this study, we evaluated the ability of Polymyxin B to neutralize the effect of LPS present as contaminant on <it>Schistosoma mansoni </it>recombinant proteins produced in <it>E. coli </it>in inducing TNF-α and IL-10. Peripheral blood mononuclear cells from individuals chronically infected with <it>S. mansoni </it>were stimulated <it>in vitro </it>with recombinant Sm22.6, Sm14 and P24 antigens (10 μg/mL) in the presence of Polymyxin B (10 μg/mL).</p> <p>Results</p> <p>The levels of cytokines were measured using ELISA. There was greater than 90 % reduction (p < 0.05) in the levels of TNF-α and IL-10 when Polymyxin B was added to the cultures stimulated with LPS. In cultures stimulated with <it>S. mansoni </it>recombinant proteins in the presence of Polymyxin B, a reduction in the levels of TNF-α and IL-10 was also observed. However, the percentage of reduction was lower when compared to the cultures stimulated with LPS, probably because these proteins are able to induce the production of these cytokines by themselves.</p> <p>Conclusion</p> <p>This study showed that Polymyxin B was able to neutralize the effect of endotoxin, as contaminant in <it>S. mansoni </it>recombinant antigens produced in <it>E. coli</it>, in inducing TNF-α and IL-10 production.</p>
ISSN:1475-2859