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Updated: May 11, 2026
Flow Cytometric Analysis for Identification of the Innate and Adaptive Immune Cells of Murine Lung
Published on: November 16, 2021
Flow cytometric analysis of macrophages and dendritic cell subsets in the mouse lung
Alexander V Misharin1, Luisa Morales-Nebreda, Gökhan M Mutlu
11 Division of Rheumatology, and.
Insights
Researchers developed a new flow cytometry panel to identify lung immune cells. This method accurately distinguishes macrophage and dendritic cell (DC) subsets, aiding research into lung injury and fibrosis.
Area of Science:
- Immunology
- Pulmonary Medicine
- Cell Biology
Background:
- Macrophages and dendritic cells (DCs) are key immune cells in lung pathology.
- Accurate identification of these subsets is crucial for understanding lung diseases.
- A standardized flow cytometry method for mouse lung immune cells was lacking.
Purpose of the Study:
- To develop a systematic flow cytometry panel for identifying lung macrophage and DC subsets.
- To characterize these immune cells in normal and injured mouse lungs.
- To assess macrophage activation states during bleomycin-induced lung injury.
Main Methods:
- Development of a novel surface marker panel for flow cytometry.
- Application of the panel to normal mouse lungs.
- Analysis of immune cells in bleomycin-induced lung injury models.
- Assessment of macrophage polarization markers.
Main Results:
- A comprehensive panel accurately identified all known lung macrophage and DC populations and their precursors.
- Alveolar macrophages showed alternatively activated markers in both acute and fibrotic injury phases.
- Classically activated macrophage markers were present only in the acute phase of injury.
Conclusions:
- The developed flow cytometry panel provides accurate and consistent identification of lung immune cell subsets.
- This tool is valuable for studying immune cell activation in normal, injured, and fibrotic lungs.
- The findings offer insights into macrophage polarization dynamics during lung injury and fibrosis.
Abstract:
The lung hosts multiple populations of macrophages and dendritic cells, which play a crucial role in lung pathology. The accurate identification and enumeration of these subsets are essential for understanding their role in lung pathology. Flow cytometry is a mainstream tool for studying the immune system. However, a systematic flow cytometric approach to identify subsets of macrophages and dendritic cells (DCs) accurately and consistently in the normal mouse lung has not been described. Here we developed a panel of surface markers and an analysis strategy that accurately identify all known populations of macrophages and DCs, and their precursors in the lung during steady-state conditions and bleomycin-induced injury. Using this panel, we assessed the polarization of lung macrophages during the course of bleomycin-induced lung injury. Alveolar macrophages expressed markers of alternatively activated macrophages during both acute and fibrotic phases of bleomycin-induced lung injury, whereas markers of classically activated macrophages were expressed only during the acute phase. Taken together, these data suggest that this flow cytometric panel is very helpful in identifying macrophage and DC populations and their state of activation in normal, injured, and fibrotic lungs.
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