Abstract
BackgroundRecombinant proteins expressed in Escherichia coli 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 Schistosoma mansoni recombinant proteins produced in E. coli in inducing TNF-α and IL-10. Peripheral blood mononuclear cells from individuals chronically infected with S. mansoni were stimulated in vitro with recombinant Sm22.6, Sm14 and P24 antigens (10 μg/mL) in the presence of Polymyxin B (10 μg/mL).ResultsThe 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 S. mansoni 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.ConclusionThis study showed that Polymyxin B was able to neutralize the effect of endotoxin, as contaminant in S. mansoni recombinant antigens produced in E. coli, in inducing TNF-α and IL-10 production.
Highlights
Recombinant proteins expressed in Escherichia coli vectors are generally contaminated with endotoxin
We evaluated the ability of Polymyxin B to neutralize the effect of LPS present as contaminant in Schistosoma mansoni recombinant proteins produced in E. coli in inducing TNF-α and IL-10 production in peripheral blood mononuclear cells (PBMC) from individuals infected with S. mansoni
Many studies have evaluated the immune response induced by recombinant proteins produced in bacteria, few remark on the potential influence of contamination with endotoxin
Summary
Recombinant proteins expressed in Escherichia coli vectors are generally contaminated with endotoxin. We evaluated the ability of Polymyxin B to neutralize the effect of LPS present as contaminant on Schistosoma mansoni recombinant proteins produced in E. coli in inducing TNF-α and IL-10. Recombinant proteins represent an important tool in the field of immunology research and have been used to understand pathogenesis and host susceptibility and may serve as components for vaccines. In general these proteins augment the specificity of diagnostic tests since they are specific peptides with known sequences. The production of most recombinant proteins is achieved through cloning into Escherichia coli vectors containing cDNA coding for the desired protein. The LPS is a component of the pathogen associated molecular (page number not for citation purposes)
Sign-in/Register to view highlights & summary Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
