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

Published: September 29, 2023

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 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 different…

Representative 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 split 1:50 into TSB with 10 µg/mL chloramphenicol and cultured. Optical density at 600 nm (OD600) was measured in regular intervals a…

Divulgazioni

The authors have nothing to disclose.

Materials

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

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Citazione di questo articolo
A Murine Skin Wound Infection Model to Study the Immune Response against a Bacterial Pathogen. J. Vis. Exp. (Pending Publication), e21695, doi: (2023).

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