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...
| Main Authors: | , , , , , |
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| Format: | Article |
| Language: | English |
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BMC
2007-01-01
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| Series: | Microbial Cell Factories |
| Online Access: | http://www.microbialcellfactories.com/content/6/1/1 |
| _version_ | 1828268094495129600 |
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| author | Pacífico Lucila G Góes Alfredo M Araujo Maria Cardoso Luciana S Oliveira Ricardo R Oliveira Sergio C |
| author_facet | Pacífico Lucila G Góes Alfredo M Araujo Maria Cardoso Luciana S Oliveira Ricardo R Oliveira Sergio C |
| author_sort | Pacífico Lucila G |
| collection | DOAJ |
| description | <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> |
| first_indexed | 2024-04-13T05:14:22Z |
| format | Article |
| id | doaj.art-11b5ea76c6c74e41aa19b39fd6092d20 |
| institution | Directory Open Access Journal |
| issn | 1475-2859 |
| language | English |
| last_indexed | 2024-04-13T05:14:22Z |
| publishDate | 2007-01-01 |
| publisher | BMC |
| record_format | Article |
| series | Microbial Cell Factories |
| spelling | doaj.art-11b5ea76c6c74e41aa19b39fd6092d202022-12-22T03:00:56ZengBMCMicrobial Cell Factories1475-28592007-01-0161110.1186/1475-2859-6-1Polymyxin B as inhibitor of LPS contamination of <it>Schistosoma mansoni </it>recombinant proteins in human cytokine analysisPacífico Lucila GGóes Alfredo MAraujo MariaCardoso Luciana SOliveira Ricardo ROliveira Sergio C<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>http://www.microbialcellfactories.com/content/6/1/1 |
| spellingShingle | Pacífico Lucila G Góes Alfredo M Araujo Maria Cardoso Luciana S Oliveira Ricardo R Oliveira Sergio C Polymyxin B as inhibitor of LPS contamination of <it>Schistosoma mansoni </it>recombinant proteins in human cytokine analysis Microbial Cell Factories |
| title | Polymyxin B as inhibitor of LPS contamination of <it>Schistosoma mansoni </it>recombinant proteins in human cytokine analysis |
| title_full | Polymyxin B as inhibitor of LPS contamination of <it>Schistosoma mansoni </it>recombinant proteins in human cytokine analysis |
| title_fullStr | Polymyxin B as inhibitor of LPS contamination of <it>Schistosoma mansoni </it>recombinant proteins in human cytokine analysis |
| title_full_unstemmed | Polymyxin B as inhibitor of LPS contamination of <it>Schistosoma mansoni </it>recombinant proteins in human cytokine analysis |
| title_short | Polymyxin B as inhibitor of LPS contamination of <it>Schistosoma mansoni </it>recombinant proteins in human cytokine analysis |
| title_sort | polymyxin b as inhibitor of lps contamination of it schistosoma mansoni it recombinant proteins in human cytokine analysis |
| url | http://www.microbialcellfactories.com/content/6/1/1 |
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