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Open Access Technical Note

A method for quantifying pulmonary Legionella pneumophila infection in mouse lungs by flow cytometry

Desmond Koon Yong Ang1*, Sze Ying Ong2, Andrew Stephen Brown1, Elizabeth Louise Hartland2 and Ian Richard van Driel1

Author Affiliations

1 Department of Biochemistry and Molecular Biology, Bio21 Molecular Science and Biotechnology Institute, The University of Melbourne, Parkville, VIC, 3010, Australia

2 Department of Microbiology and Immunology, The University of Melbourne, Parkville, VIC, 3010, Australia

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BMC Research Notes 2012, 5:448  doi:10.1186/1756-0500-5-448

Published: 20 August 2012

Abstract

Background

Pulmonary load of Legionella pneumophila in mice is normally determined by counting serial dilutions of bacterial colony forming units (CFU) on agar plates. This process is often tedious and time consuming. We describe a novel, rapid and versatile flow cytometric method that detects bacteria phagocytosed by neutrophils.

Findings

Mice were infected with L. pneumophila via intratracheal or intranasal administration. At various times after bacteria inoculation, mouse lungs were harvested and analysed concurrently for bacterial load by colony counting and flow cytometry analysis. The number of L. pneumophila-containing neutrophils correlated strongly with CFU obtained by bacteriological culture.

Conclusions

This technique can be utilised to determine pulmonary bacterial load and may be used in conjunction with other flow cytometric based analyses of the resulting immune response.

Keywords:
Legionella pneumophila; Legionnaires’ disease; Neutrophil; Flow cytometry; Mouse infection model