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

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Samenvatting

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 California, Davis in groups prior to surgery and single-housed following surgery. Use mice between the ages of 10-16 weeks.

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Resultaten

SH1000 growth curve; OD600 vs CFU counts; linear correlation graph; R²=0.9214; equation shown.
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/m.......