Flow Cytometry of Lung and Bronchoalveolar Lavage Fluid Cells from Mice Challenged with Fluorescent Aspergillus
Anupam Jhingran1, Shinji Kasahara1, Tobias M Hohl1,2
1Infectious Disease Service, Department of Medicine, Memorial Sloan Kettering Cancer Center, New York, USA.
Abstract:
Aspergillus fumigatus is a ubiquitous fungal pathogen that forms airborne conidia. The process of restricting conidial germination into hyphae by lung leukocytes is critical in determining infectious outcomes. Tracking the outcome of conidia-host cell encounters in vivo is technically challenging and an obstacle to understanding the molecular and cellular basis of antifungal immunity in the lung. Here, we describe a method that utilizes a genetically engineered Aspergillus strain [called FLARE (Jhingran et al., 2012; Espinosa et al., 2014; Heung et al., 2015)] to monitor conidial phagocytosis and killing by leukocytes within the lung environment at single encounter resolution.
Insights
Researchers developed a novel method using genetically engineered Aspergillus fumigatus to track fungal spore interactions with immune cells in the lungs. This technique allows detailed observation of phagocytosis and killing by leukocytes, advancing antifungal immunity research.
Area of Science:
- Medical Mycology
- Immunology
- Infectious Diseases
Background:
- Aspergillus fumigatus is a common airborne fungal pathogen.
- Lung leukocyte interaction with fungal conidia is crucial for controlling infection.
- Studying these interactions in vivo is difficult, hindering understanding of antifungal immunity.
Purpose of the Study:
- To develop a new method for observing Aspergillus fumigatus conidia-host cell interactions in the lung.
- To enable single-encounter resolution tracking of phagocytosis and killing by leukocytes.
- To advance the understanding of the molecular and cellular basis of lung antifungal immunity.
Main Methods:
- Utilized a genetically engineered Aspergillus fumigatus strain (FLARE).
- Developed a method to monitor conidial interactions with leukocytes in the lung environment.
- Enabled tracking at single encounter resolution.
Main Results:
- Successfully monitored conidial phagocytosis by lung leukocytes.
- Enabled real-time observation of conidia-leukocyte encounters in vivo.
- Provided a tool for high-resolution analysis of antifungal immune responses.
Conclusions:
- The FLARE strain and developed method offer a powerful tool for studying host-pathogen interactions in the lung.
- This approach facilitates detailed investigation into the mechanisms of antifungal immunity.
- Improved understanding of how lung leukocytes combat Aspergillus fumigatus infections.
