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Analyzing Macrophage Infection at the Organ Level

Part of the Methods in Molecular Biology book series (MIMB,volume 2414)

Abstract

Classical in vivo infection models are oftentimes associated with speculation due to the many physiological factors that are unseen or not accounted for when analyzing experimental outputs, especially when solely utilizing the classic approach of tissue-derived colony-forming unit (CFU) enumeration. To better understand the steps and natural progression of bacterial infection, the pathophysiology of individual organs with which the bacteria interact in their natural course of infection must be considered. In this case, it is not only important to isolate organs as much as possible from additional physiological processes, but to also consider the dynamics of the bacteria at the cellular level within these organs of interest. Here, we describe in detail two models, ex vivo porcine liver and spleen coperfusion and a murine infection model, and the numerous associated experimental outputs produced by these models that can be taken and used together to investigate the pathogen–host interactions within tissues in depth.

Key words

  • Ex vivo perfusion
  • Murine infection model
  • Immunohistochemistry
  • Confocal microscopy
  • Fiji
  • InForm
  • Image analysis
  • Correlates of protection

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Acknowledgements

We thank John Isherwood and Rohan Kumar for help with the perfusion of the porcine organs at explant, the staff of Joseph Morris Butchers, and Sarah Glenn and the staff of the Leicester Preclinical Research Facility for support with the mouse experiments. The grant was in part supported by a collaboration agreement with the University of Oxford and grants from the MRC MR/M003078/1 and BBSRC BB/S507052/1 to MRO. ZJ is funded by BBSRC BB/S507052/1.

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Correspondence to Marco R. Oggioni .

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Hames, R.G. et al. (2022). Analyzing Macrophage Infection at the Organ Level. In: Bidmos, F., Bossé, J., Langford, P. (eds) Bacterial Vaccines. Methods in Molecular Biology, vol 2414. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1900-1_22

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  • DOI: https://doi.org/10.1007/978-1-0716-1900-1_22

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-1899-8

  • Online ISBN: 978-1-0716-1900-1

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