New Mouse Model for Chronic Infections by Gram-Negative Bacteria Enabling the Study of Anti-Infective Efficacy and Host-Microbe Interactions

• Published in: mBio 2017-02, Vol.8 (1)
• Main Authors: Pletzer, Daniel, Mansour, Sarah C, Wuerth, Kelli, Rahanjam, Negin, Hancock, Robert E W
• Format: Article
• Language: English
• Subjects: Abscess - drug therapy; Abscess - microbiology; Abscess - pathology; Animals; Anti-Bacterial Agents - therapeutic use; Chronic Disease; Disease Models, Animal; Gram-Negative Bacteria - pathogenicity; Gram-Negative Bacterial Infections - drug therapy; Gram-Negative Bacterial Infections - microbiology; Gram-Negative Bacterial Infections - pathology; Host-Pathogen Interactions; Mice; Mice, Inbred C57BL; Skin Diseases, Bacterial - drug therapy; Skin Diseases, Bacterial - microbiology; Skin Diseases, Bacterial - pathology; Treatment Outcome
• ISSN: 2161-2129; 2150-7511
• DOI: 10.1128/mBio.00140-17

Only a few, relatively cumbersome animal models enable long-term Gram-negative bacterial infections that mimic human situations, where untreated infections can last for weeks. Here, we describe a simple murine cutaneous abscess model that enables chronic or progressive infections, depending on the subcutaneously injected bacterial strain. In this model, cystic fibrosis epidemic isolate LESB58 caused localized high-density skin and soft tissue infections and necrotic skin lesions for up to 10 days but did not disseminate in either CD-1 or C57BL/6 mice. The model was adapted for use with four major Gram-negative nosocomial pathogens, , , , and This model enabled noninvasive imaging and tracking of -tagged bacteria, the influx of activated neutrophils, and production of reactive oxygen-nitrogen species at the infection site. Screening antimicrobials against high-density infections showed that local but not intravenous administration of gentamicin, ciprofloxacin, and meropenem significantly but incompletely reduced bacterial counts and superficial tissue dermonecrosis. Bacterial RNA isolated from the abscess tissue revealed that genes involved in iron uptake, toxin production, surface lipopolysaccharide regulation, adherence, and lipase production were highly upregulated whereas phenazine production and expression of global activator were downregulated. The model was validated for studying virulence using mutants of more-virulent strain PA14. Thus, mutants defective in flagella or motility, type III secretion, or siderophore biosynthesis were noninvasive and suppressed dermal necrosis in mice, while a strain with a mutation in the gene encoding a sensor kinase showed enhanced invasiveness and mortality in mice compared to controls infected with wild-type PA14. More than two-thirds of hospital infections are chronic or high-density biofilm infections and difficult to treat due to adaptive, multidrug resistance. Unfortunately, current models of chronic infection are technically challenging and difficult to track without sacrificing animals. Here we describe a model of chronic subcutaneous infection and abscess formation by medically important nosocomial Gram-negative pathogens that is simple and can be used for tracking infections by imaging, examining pathology and immune responses, testing antimicrobial treatments suitable for high-density bacterial infections, and studying virulence. We propose that this mouse model can be a game changer for modeling hard-to-