Potential role for toll-like receptor 4 in mediating Escherichia coli maltose-binding protein activation of dendritic cells

Stefan Fernandez*, Dupeh R. Palmer, Monika Simmons, Peifang Sun, John Bisbing, Sasha McClain, Sachin Mani, Timothy Burgess, Vicky Gunther, Wellington Sun

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

34 Scopus citations

Abstract

The Escherichia coli maltose-binding protein (MBP) is used to increase the stability and solubility of proteins in bacterial protein expression systems and is increasingly being used to facilitate the production and delivery of subunit vaccines against various pathogenic bacteria and viruses. The MBP tag is presumed inert, with minimum effects on the bioactivity of the tagged protein or its biodistribution. However, few studies have characterized the immunological attributes of MBP. Here, we analyze the phenotypic and functional outcomes of MBP-treated dendritic cells (DCs) and show that MBP induces DC activation and production of proinflammatory cytokines (interleukin-1β [IL-1β], IL-6, IL-8, tumor necrosis factor alpha, and IL-12p70) within 24 h and strongly increases IκB phosphorylation in treated cells. Interestingly, phosphorylation of IκB was largely abrogated by the addition of anti-human Toll-like receptor 4 (TLR4) antibodies, indicating that MBP activates signaling for DC maturation via TLR4. Consistent with this hypothesis, MBP activated the TLR4-expressing cell line 293-hTLR4A but not control cultures to secrete IL-8. The observed data were independent of lipopolysaccharide contamination and support a role for TLR4 in mediating the effects of MBP. These results provide insight into a mechanism by which MBP might enhance immune responses to vaccine fusion proteins.

Original languageEnglish
Pages (from-to)1359-1363
Number of pages5
JournalInfection and Immunity
Volume75
Issue number3
DOIs
StatePublished - Mar 2007
Externally publishedYes

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