title: Interrupting Vibrio cholerae infection of human epithelial cells with engineered commensal bacterial signaling.
abstract: Vibrio cholerae El Tor serotypes are largely responsible for outbreaks of cholera in the developing world. The infection cycle for some strains of V. cholerae is coordinated, at least in part, through quorum sensing. That is, the expression of virulence genes depends on the concentration of V. cholerae autoinducers cholera autoinducer 1 (CAI-1) and autoinducer 2 (AI-2). High concentrations of CAI-1 and AI-2 have been shown previously to inhibit virulence gene expression. We have demonstrated here that a commensal bacterium, E. coli Nissle 1917 (Nissle), can be engineered to express CAI-1 (Nissle expresses AI-2 natively) and effectively interrupt V. cholerae virulence. We engineered Nissle to express CAI-1 under control of the lac promoter, and demonstrated inhibition of V. cholerae expression of cholera toxin (CT, as indicated by presence of the CT subunit B (CTB)) and of the toxin co-regulated pilus (TCP, as indicated by the relative transcript of TCP subunit A (TCPA)) in both monocultures of V. cholerae and co-cultures with epithelial cells, Nissle, and V. cholerae. In the model system of Caco-2 epithelia incubated with V. cholerae, we demonstrated that co-cultures with Nissle expressing CAI-1 activity reduced CTB binding to Caco-2 cells by 63% over co-cultures with wild-type Nissle. Further, cultures with Nissle expressing CAI-1 had significantly lower TCPA transcription than controls with wild-type Nissle. These results represent a significant step towards a prophylactic method for combating enteric disease through engineered quorum signaling within a commensal bacterial strain.
authors.affiliation: Department of Biological and Environmental Engineering, Cornell University, Ithaca, New York 14853, USA.
authors.forename: Faping
authors.initials: F
authors.lastname: Duan
authors.forename: John C
authors.initials: JC
authors.lastname: March
mesh_terms: D002478:Cells, Cultured; D004847:Epithelial Cells / Q000382:microbiology*; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D007659:Ketones / Q000378:metabolism*; D053038:Quorum Sensing / Q000502:physiology*; D014734:Vibrio cholerae / Q000254:growth & development*
chemical_list: C525620:3-hydroxytridecan-4-one; D007659:Ketones