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Published on: June 19, 2013
Labelling of Candida auris Cell Walls to Examine Levels of PAMP Exposure by Flow Cytometry and Fluorescence
Chloe Pelletier1,2, Alexander Lorenz3
1Institute of Medical Sciences (IMS), University of Aberdeen, Aberdeen, UK.
Abstract:
Pathogen-associated molecular patterns (PAMPs) of the fungal cell wall are primary targets for the innate immune system of animals. Therefore, characterizing PAMP exposure of fungal pathogens helps to elucidate how they interact with their hosts at a molecular level. Fluorescent labelling can be used to monitor exposure of multiple fungal cell wall PAMPs in a single experiment. Here, we describe a protocol to simultaneously label chitin, mannan, and β-1,3-glucan in Candida auris to study these PAMPs by fluorescence microscopy and allow high-throughput examination of their exposure by flow cytometry.
Insights
This study presents a novel method for simultaneously labeling fungal cell wall components like chitin, mannan, and beta-1,3-glucan in Candida auris. This technique aids in understanding pathogen-host interactions at a molecular level.
Area of Science:
- Mycology
- Immunology
- Biochemistry
Background:
- Pathogen-associated molecular patterns (PAMPs) on fungal cell walls are crucial targets for animal innate immunity.
- Understanding PAMP exposure is key to elucidating fungal pathogen-host molecular interactions.
Purpose of the Study:
- To develop a protocol for simultaneous fluorescent labeling of multiple fungal cell wall PAMPs.
- To enable detailed study of PAMP exposure in Candida auris using advanced microscopy techniques.
Main Methods:
- Simultaneous fluorescent labeling of chitin, mannan, and β-1,3-glucan in Candida auris.
- Utilizing fluorescence microscopy for detailed PAMP visualization.
- Employing flow cytometry for high-throughput PAMP exposure analysis.
Main Results:
- Successful simultaneous labeling of key fungal cell wall PAMPs.
- Demonstrated feasibility of studying PAMP exposure in Candida auris.
- Established a method for high-throughput analysis of PAMP exposure.
Conclusions:
- The developed protocol allows for comprehensive analysis of fungal cell wall PAMP exposure.
- This method facilitates a deeper understanding of Candida auris interactions with host immune systems.
- Fluorescent labeling offers a powerful tool for studying fungal pathogen virulence factors.
