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Assessing bacterial invasion of cardiac cells in culture and heart colonization in infected mice using Listeria monocytogenes.

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journal contribution
posted on 13.05.2017, 00:00 by PD McMullen, NE Freitag
Listeria monocytogenes is a Gram-positive facultative intracellular pathogen that is capable of causing serious invasive infections in immunocompromised patients, the elderly, and pregnant women. The most common manifestations of listeriosis in humans include meningitis, encephalitis, and fetal abortion. A significant but much less documented sequelae of invasive L. monocytogenes infection involves the heart. The death rate from cardiac illness can be up to 35% despite treatment, however very little is known regarding L. monocytogenes colonization of cardiac tissue and its resultant pathologies. In addition, it has recently become apparent that subpopulations of L. monocytogenes have an enhanced capacity to invade and grow within cardiac tissue. This protocol describes in detail in vitro and in vivo methods that can be used for assessing cardiotropism of L. monocytogenes isolates. Methods are presented for the infection of H9c2 rat cardiac myoblasts in tissue culture as well as for the determination of bacterial colonization of the hearts of infected mice. These methods are useful not only for identifying strains with the potential to colonize cardiac tissue in infected animals, but may also facilitate the identification of bacterial gene products that serve to enhance cardiac cell invasion and/or drive changes in heart pathology. These methods also provide for the direct comparison of cardiotropism between multiple L. monocytogenes strains.

Funding

Grant Support: AI099339/AI/NIAID NIH HHS/United States AI41816/AI/NIAID NIH HHS/United States F31AI094886-01/AI/NIAID NIH HHS/United States R21 AI099339/AI/NIAID NIH HHS/United States

History

Publisher Statement

This is the copy of an article published in the Journal of Visualized Experiments © 2015 Journal of Visualized Experiments Publications.

Publisher

Journal of Visualized Experiments

issn

1940-087X

Issue date

27/05/2015