A Murine Skin Wound Infection Model to Study the Immune Response against a Bacterial Pathogen

Abstract

Source: Anderson, L. S., et al. A Mouse Model to Assess Innate Immune Response to Staphylococcus aureus Infection. J. Vis. Exp. (2019).

This video demonstrates a technique to generate a murine model of skin wound infection. Upon infecting mice — engineered to express a fluorescent protein in neutrophils — with a genetically modified bioluminescent Staphylococcus aureus — a pathogenic bacteria. The bacterial burden at the infection site and the neutrophil recruitment dynamics can be studied using whole animal imaging.

Protocol

All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board.

1. Mouse Source and Housing

1. Derive LysM-eGFP (enhanced green fluorescent protein (eGFP) inserted in the Lysozyme M (LysM) promoter region) mice on a C57BL/6J genetic background as described previously. Derive LysM-EGFP×MyD88^-/- mice by crossing LysM-EGFP mice with MyD88^-/- (Myeloid differentiation primary response 88 protein) mice on a C57BL/6J background.
2. House mice in a vivarium. For these studies, animals were housed at the Univer....

Results

Figure 1: Correlation between OD₆₀₀ and CFU counts for ALC2906. 3-4 ALC2906 SH1000 colonies were picked from an agar plate and transferred into TSB with 10 µg/mL chloramphenicol for overnight culture. The next day, the suspension was spli.......

Materials

List of materials used in this article

Name	Company	Catalog Number	Comments
14 mL Polypropylene Round-Bottom Tube	Falcon	352059
6mm Disposable Biopsy Punch	Integra Miltex	33-36
Bioluminescent S. aureus	Lloyd Miller, Johns Hopkins	ALC 2906 SH1000
Bovine Blood Agar, 5%, Hardy Diagnostics	VWR	10118-938
Buprenoprhine hydrochloride injectable	Western Medical Supply	7292	0.3 mg/mL
C57BL/6J Mice	Jackson Labratory	664
Chloramphenicol (crystalline powder)	Fisher BioReagents	BP904-100
DPBS (1X)	Gibco	14190-144
Insulin Syringes	Becton, Dickson and Company	329461	.35 mm (28 G) x 12.7 mm (1/2'')
IVIS Spectrum In Vivo Imaging System	Perkin Elmer	124262
Living Image Software – IVIS Spectrum Series	Perkin Elmer	128113
LysM-eGFP Mice	Thomas Graff Albert Einstein College of New York	NA
Microvolume Spectrophotometer	ThermoFisher Scientific	ND-2000
MyD88 KO Mice	Jackson Labratory	9088
Non-woven sponges	AMD- Ritmed Inc	A2101-CH	5 cm x 5 cm
Povidone Iodine 10% Solution	Aplicare	697731
Tryptic Soy Broth	Becton, Dickson and Company	211825