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Updated: Jun 3, 2026
Viability Assay of Trichoderma stromaticum Conidia Inside Human Peripheral Blood Mononuclear-Derived Macrophages
Published on: October 20, 2023
In vivo and ex vivo protocols for measuring the killing of extracellular pathogens by macrophages
1Infection Immunology Research Group, Helmholtz Centre for Infection Research, Braunschweig, Germany.
Abstract:
This unit describes a series of in vivo/ex vivo combined protocols for investigating the interactions (adhesion, phagocytosis, and killing) of extracellular bacteria with peritoneal murine macrophages. It includes steps needed for in vivo infection of murine peritoneal macrophages after intraperitoneal inoculation with the pathogen of interest, as well as the measurement of bacteria associated with or truly internalized by these phagocytic cells. Several protocols for the ex vivo measurement of the ability of peritoneal macrophages to kill the microorganisms that have been ingested during the in vivo infection assay are included.
Insights
This study details combined in vivo/ex vivo methods to analyze how extracellular bacteria interact with mouse macrophages. It covers bacterial adhesion, phagocytosis, and killing by these immune cells following infection.
Area of Science:
- Immunology
- Microbiology
- Cell Biology
Background:
- Peritoneal macrophages are key immune cells involved in clearing bacterial infections.
- Understanding bacterial-macrophage interactions is crucial for developing effective antimicrobial strategies.
- Existing methods may not fully capture the dynamic interplay between bacteria and macrophages in vivo and ex vivo.
Purpose of the Study:
- To describe integrated in vivo and ex vivo protocols for studying extracellular bacterial interactions with murine peritoneal macrophages.
- To provide methodologies for assessing bacterial adhesion, phagocytosis, and killing by macrophages.
- To enable comprehensive analysis of macrophage-mediated bacterial clearance.
Main Methods:
- In vivo intraperitoneal inoculation of mice with pathogenic bacteria.
- Isolation and quantification of peritoneal macrophages from infected mice.
- Assessment of bacterial association (adhesion) and internalization (phagocytosis) by macrophages.
- Ex vivo assays to measure the killing capacity of macrophages against ingested bacteria.
Main Results:
- Established protocols for simultaneous in vivo infection and ex vivo analysis of bacterial-macrophage interactions.
- Quantification of bacterial adhesion and phagocytosis rates.
- Measurement of macrophage-mediated bacterial killing efficiency.
- Detailed procedures for analyzing immune cell-pathogen dynamics.
Conclusions:
- The described combined in vivo/ex vivo approach offers a robust platform for dissecting bacterial-macrophage interactions.
- These protocols facilitate the evaluation of bacterial virulence factors and host immune responses.
- This methodology aids in the study of host-pathogen dynamics and the development of novel therapeutic interventions.
