Lien vers Pubmed [PMID] – 19444238
Lien DOI – 10.1038/nprot.2009.66
Nat Protoc 2009 ; 4(6): 799-810
Listeria monocytogenes causes listeriosis, a human foodborne infection leading to gastroenteritis, meningoencephalitis and maternofetal infections. InlA and InlB, two L. monocytogenes surface proteins, interact with their respective receptors E-cadherin and Met and mediate bacterial entry into human cultured cells. Here, we present protocols for studying listeriosis in three complementary animal models: (i) the human E-cadherin (hEcad) transgenic mouse line; (ii) the knock-in E16P mouse line; and (iii) the gerbil, in which both InlA-E-cadherin and InlB-Met species-specific interactions occur as in humans. Two routes of infection are described: oral inoculation, the natural route for infection; and intravenous inoculation that bypasses the intestinal barrier. We describe how to monitor L. monocytogenes infection, both qualitatively by imaging techniques and quantitatively by bacterial enumeration. The advantage of these methods over the classical intravenous inoculation of L. monocytogenes in wild-type mice (in which the InlA-E-cadherin interaction does not occur) is that it allows the pathophysiology of listeriosis to be studied in animal models relevant to humans, as they are permissive to the interactions that are thought to mediate L. monocytogenes crossing of human host barriers. The whole procedure (inoculation, in vivo imaging, bacterial enumeration, histopathology) takes one full week to complete, including 3 d of actual experiments.