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Updated: Oct 18, 2025
Fluorescence-mediated Tomography for the Detection and Quantification of Macrophage-related Murine Intestinal Inflammation
Published on: December 15, 2017
Characterization of macrophage infiltration and polarization by double fluorescence immunostaining in mouse colonic
María López Chiloeches1,2, Anna Bergonzini1,2, Teresa Frisan1,2
1Department of Molecular Biology, Umeå University, 90736 Umeå, Sweden.
Abstract:
We recently characterized the association between DNA damage and immunoresponse in vivo in colonic mucosa of mice infected with a Salmonella Typhimurium strain expressing a genotoxin, known as typhoid toxin. In this protocol, we describe how to assess the extent and features of infiltrating macrophages by double immunofluorescence. Total macrophage population was determined using an F4/80 antibody, whereas the specific M2-like population was assessed using a CD206 antibody. For complete details on the use and execution of this protocol, please refer to Martin et al. (2021).
Insights
This study details a method to analyze macrophage responses to DNA damage in mice infected with Salmonella Typhimurium. Researchers used double immunofluorescence to identify macrophage populations, aiding in understanding immune responses to genotoxins.
Area of Science:
- Immunology
- Microbiology
- Molecular Biology
Background:
- DNA damage and immune responses are crucial in infectious diseases.
- Salmonella Typhimurium infection can induce genotoxicity via typhoid toxin.
- Understanding macrophage infiltration is key to studying host-pathogen interactions.
Purpose of the Study:
- To describe a protocol for assessing macrophage populations in the colonic mucosa.
- To investigate the association between DNA damage and immune response in vivo.
- To quantify total and M2-like macrophages during Salmonella infection.
Main Methods:
- Utilizing double immunofluorescence staining in mouse colonic mucosa.
- Employing F4/80 antibody to identify total macrophages.
- Using CD206 antibody to assess the M2-like macrophage subset.
Main Results:
- The protocol allows for the characterization of infiltrating macrophages.
- Quantification of F4/80+ and CD206+ cells provides insights into immune cell dynamics.
- The method facilitates the study of immune responses to genotoxic bacterial infections.
Conclusions:
- This protocol provides a reliable method for studying macrophage responses in the context of DNA damage and Salmonella infection.
- The findings contribute to understanding the interplay between genotoxicity and host immunity.
- Further research can build upon this methodology to explore therapeutic strategies.
