Monitoring Neutrophil Recruitment Dynamics and Bacterial Burden at the Infection Site in a Murine Model

Published: August 31, 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 in vivo imaging to study the neutrophil recruitment dynamics at an infection site in an immunocompromised, transgenic mouse with GFP-expressing neutrophils. It involves injecting luminescent bacteria to cause an infection. This leads to neutrophil recruitment to the infection site, whose tracking is done by 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 mice on a C57BL/6J genetic background. Derive LysM-EGFP×MyD88-/- mice by crossing LysM-EGFP mice with MyD88-/- mice on a C57BL/6J background. House mice in a vivarium. For these studies, animals were housed at the University of Cali…

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 after 2 hours using a sp…

開示

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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記事を引用
Monitoring Neutrophil Recruitment Dynamics and Bacterial Burden at the Infection Site in a Murine Model. J. Vis. Exp. (Pending Publication), e21603, doi: (2023).

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