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Citrobacter rodentium mouse model of bacterial infection

Nature Protocols volume 11, pages 1851–1876 (2016)Cite this article

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Abstract

Infection of mice with Citrobacter rodentium is a robust model to study bacterial pathogenesis, mucosal immunology, the health benefits of probiotics and the role of the microbiota during infection. C. rodentium was first isolated by Barthold from an outbreak of mouse diarrhea in Yale University in 1972 and was 'rediscovered' by Falkow and Schauer in 1993. Since then the use of the model has proliferated, and it is now the gold standard for studying virulence of the closely related human pathogens enteropathogenic and enterohemorrhagic Escherichia coli (EPEC and EHEC, respectively). Here we provide a detailed protocol for various applications of the model, including bacterial growth, site-directed mutagenesis, mouse inoculation (from cultured cells and after cohabitation), monitoring of bacterial colonization, tissue extraction and analysis, immune responses, probiotic treatment and microbiota analysis. The main protocol, from mouse infection to clearance and analysis of tissues and host responses, takes ∼5 weeks to complete.

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Figure 1: Protocol workflow.
Figure 2: Schematic representation of the generation of C. rodentium chromosomal deletion mutant.
Figure 3: In vitro organ cultures.
Figure 7: H&E and immunofluorescence staining.
Figure 6: Recruitment of immune cells during C. rodentium infection.
Figure 4: Attaching and effacing lesions.
Figure 5: C. rodentium colonization dynamics and imaging.

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Acknowledgements

Many of the protocols described in this manuscript were developed through grants from the Wellcome Trust and the Medical Research Council (MRC).

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  1. Department of Life Sciences, MRC Centre for Molecular Bacteriology and Infection, Imperial College, London, UK

    Valerie F Crepin, James W Collins, Maryam Habibzay & Gad Frankel

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  1. Valerie F Crepin
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V.F.C., J.W.C., M.H. and G.F. wrote the paper.

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Correspondence to Gad Frankel.

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Crepin, V., Collins, J., Habibzay, M. et al. Citrobacter rodentium mouse model of bacterial infection. Nat Protoc 11, 1851–1876 (2016). https://doi.org/10.1038/nprot.2016.100

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