Open Access Highly Accessed Methodology article

Bioluminescence imaging to track bacterial dissemination of Yersinia pestis using different routes of infection in mice

Rodrigo J Gonzalez1, Eric H Weening2, Richard Frothingham34, Gregory D Sempowski35 and Virginia L Miller12*

Author Affiliations

1 Department of Microbiology and Immunology, University of North Carolina, Chapel Hill, NC, USA

2 Department of Genetics, University of North Carolina, Chapel Hill, NC, USA

3 Department of Medicine and Duke Human Vaccine Institute, Duke University Medical Center, Durham, NC, USA

4 Department of Molecular Genetics and Microbiology, Duke University Medical Center, Durham, NC, USA

5 Department of Pathology, Duke University Medical Center, Durham, NC, USA

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BMC Microbiology 2012, 12:147  doi:10.1186/1471-2180-12-147

Published: 24 July 2012

Abstract

Background

Plague is caused by Yersinia pestis, a bacterium that disseminates inside of the host at remarkably high rates. Plague bacilli disrupt normal immune responses in the host allowing for systematic spread that is fatal if left untreated. How Y. pestis disseminates from the site of infection to deeper tissues is unknown. Dissemination studies for plague are typically performed in mice by determining the bacterial burden in specific organs at various time points. To follow bacterial dissemination during plague infections in mice we tested the possibility of using bioluminescence imaging (BLI), an alternative non-invasive approach. Fully virulent Y. pestis was transformed with a plasmid containing the luxCDABE genes, making it able to produce light; this lux-expressing strain was used to infect mice by subcutaneous, intradermal or intranasal inoculation.

Results

We successfully obtained images from infected animals and were able to follow bacterial dissemination over time for each of the three different routes of inoculation. We also compared the radiance signal from animals infected with a wild type strain and a Ī”caf1Ī”psaA mutant that we previously showed to be attenuated in colonization of the lymph node and systemic dissemination. Radiance signals from mice infected with the wild type strain were larger than values obtained from mice infected with the mutant strain (linear regression of normalized values, Pā€‰<ā€‰0.05).

Conclusions

We demonstrate that BLI is useful for monitoring dissemination from multiple inoculation sites, and for characterization of mutants with defects in colonization or dissemination.

Keywords:
Plague; Bioluminescence; In vivo imaging; Bacterial dissemination