Characterisation of <i>Aspergillus fumigatus</i> Endocytic Trafficking within Airway Epithelial Cells Using High-Resolution Automated Quantitative Confocal Microscopy
The precise characterization of the mechanisms modulating <i>Aspergillus fumigatus</i> survival within airway epithelial cells has been impaired by the lack of live-cell imaging technologies and user-friendly quantification approaches. Here we described the use of an automated image anal...
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| Format: | Article |
| Language: | English |
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MDPI AG
2021-06-01
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| Series: | Journal of Fungi |
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| Online Access: | https://www.mdpi.com/2309-608X/7/6/454 |
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| author | Nagwa Ben-Ghazzi Sergio Moreno-Velásquez Constanze Seidel Darren Thomson David W. Denning Nick D. Read Paul Bowyer Sara Gago |
| author_facet | Nagwa Ben-Ghazzi Sergio Moreno-Velásquez Constanze Seidel Darren Thomson David W. Denning Nick D. Read Paul Bowyer Sara Gago |
| author_sort | Nagwa Ben-Ghazzi |
| collection | DOAJ |
| description | The precise characterization of the mechanisms modulating <i>Aspergillus fumigatus</i> survival within airway epithelial cells has been impaired by the lack of live-cell imaging technologies and user-friendly quantification approaches. Here we described the use of an automated image analysis pipeline to estimate the proportion of <i>A. fumigatus</i> spores taken up by airway epithelial cells, those contained within phagolysosomes or acidified phagosomes, along with the fungal factors contributing to these processes. Coupling the use of fluorescent <i>A. fumigatus</i> strains and fluorescent epithelial probes targeting lysosomes, acidified compartments and cell membrane, we found that both the efficacy of lysosome recruitment to phagosomes and phagosome acidification determines the capacity of airway epithelial cells to contain <i>A. fumigatus</i> growth. Overall, the capability of the airway epithelium to prevent <i>A. fumigatus</i> survival was higher in bronchial epithelial than alveolar epithelial cells. Certain <i>A. fumigatus</i> cell wall mutants influenced phagosome maturation in airway epithelial cells. Taken together, this live-cell 4D imaging approach allows observation and measurement of the very early processes of <i>A. fumigatus</i> interaction within live airway epithelial monolayers. |
| first_indexed | 2024-03-10T10:38:32Z |
| format | Article |
| id | doaj.art-e6d5ce42797b4c71b4b549f3ff8fa475 |
| institution | Directory Open Access Journal |
| issn | 2309-608X |
| language | English |
| last_indexed | 2024-03-10T10:38:32Z |
| publishDate | 2021-06-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Journal of Fungi |
| spelling | doaj.art-e6d5ce42797b4c71b4b549f3ff8fa4752023-11-21T23:08:14ZengMDPI AGJournal of Fungi2309-608X2021-06-017645410.3390/jof7060454Characterisation of <i>Aspergillus fumigatus</i> Endocytic Trafficking within Airway Epithelial Cells Using High-Resolution Automated Quantitative Confocal MicroscopyNagwa Ben-Ghazzi0Sergio Moreno-Velásquez1Constanze Seidel2Darren Thomson3David W. Denning4Nick D. Read5Paul Bowyer6Sara Gago7Manchester Fungal Infection Group, Division of Infection, Immunity and Respiratory Medicine, Faculty of Biology, Medicine and Health, School of Biological Sciences, The University of Manchester, 2nd Floor, 46 Grafton Street, Manchester M13 9NT, UKManchester Fungal Infection Group, Division of Infection, Immunity and Respiratory Medicine, Faculty of Biology, Medicine and Health, School of Biological Sciences, The University of Manchester, 2nd Floor, 46 Grafton Street, Manchester M13 9NT, UKManchester Fungal Infection Group, Division of Infection, Immunity and Respiratory Medicine, Faculty of Biology, Medicine and Health, School of Biological Sciences, The University of Manchester, 2nd Floor, 46 Grafton Street, Manchester M13 9NT, UKManchester Fungal Infection Group, Division of Infection, Immunity and Respiratory Medicine, Faculty of Biology, Medicine and Health, School of Biological Sciences, The University of Manchester, 2nd Floor, 46 Grafton Street, Manchester M13 9NT, UKManchester Fungal Infection Group, Division of Infection, Immunity and Respiratory Medicine, Faculty of Biology, Medicine and Health, School of Biological Sciences, The University of Manchester, 2nd Floor, 46 Grafton Street, Manchester M13 9NT, UKManchester Fungal Infection Group, Division of Infection, Immunity and Respiratory Medicine, Faculty of Biology, Medicine and Health, School of Biological Sciences, The University of Manchester, 2nd Floor, 46 Grafton Street, Manchester M13 9NT, UKManchester Fungal Infection Group, Division of Infection, Immunity and Respiratory Medicine, Faculty of Biology, Medicine and Health, School of Biological Sciences, The University of Manchester, 2nd Floor, 46 Grafton Street, Manchester M13 9NT, UKManchester Fungal Infection Group, Division of Infection, Immunity and Respiratory Medicine, Faculty of Biology, Medicine and Health, School of Biological Sciences, The University of Manchester, 2nd Floor, 46 Grafton Street, Manchester M13 9NT, UKThe precise characterization of the mechanisms modulating <i>Aspergillus fumigatus</i> survival within airway epithelial cells has been impaired by the lack of live-cell imaging technologies and user-friendly quantification approaches. Here we described the use of an automated image analysis pipeline to estimate the proportion of <i>A. fumigatus</i> spores taken up by airway epithelial cells, those contained within phagolysosomes or acidified phagosomes, along with the fungal factors contributing to these processes. Coupling the use of fluorescent <i>A. fumigatus</i> strains and fluorescent epithelial probes targeting lysosomes, acidified compartments and cell membrane, we found that both the efficacy of lysosome recruitment to phagosomes and phagosome acidification determines the capacity of airway epithelial cells to contain <i>A. fumigatus</i> growth. Overall, the capability of the airway epithelium to prevent <i>A. fumigatus</i> survival was higher in bronchial epithelial than alveolar epithelial cells. Certain <i>A. fumigatus</i> cell wall mutants influenced phagosome maturation in airway epithelial cells. Taken together, this live-cell 4D imaging approach allows observation and measurement of the very early processes of <i>A. fumigatus</i> interaction within live airway epithelial monolayers.https://www.mdpi.com/2309-608X/7/6/454<i>Aspergillus fumigatus</i>airway epithelial cellsphagocytosis |
| spellingShingle | Nagwa Ben-Ghazzi Sergio Moreno-Velásquez Constanze Seidel Darren Thomson David W. Denning Nick D. Read Paul Bowyer Sara Gago Characterisation of <i>Aspergillus fumigatus</i> Endocytic Trafficking within Airway Epithelial Cells Using High-Resolution Automated Quantitative Confocal Microscopy Journal of Fungi <i>Aspergillus fumigatus</i> airway epithelial cells phagocytosis |
| title | Characterisation of <i>Aspergillus fumigatus</i> Endocytic Trafficking within Airway Epithelial Cells Using High-Resolution Automated Quantitative Confocal Microscopy |
| title_full | Characterisation of <i>Aspergillus fumigatus</i> Endocytic Trafficking within Airway Epithelial Cells Using High-Resolution Automated Quantitative Confocal Microscopy |
| title_fullStr | Characterisation of <i>Aspergillus fumigatus</i> Endocytic Trafficking within Airway Epithelial Cells Using High-Resolution Automated Quantitative Confocal Microscopy |
| title_full_unstemmed | Characterisation of <i>Aspergillus fumigatus</i> Endocytic Trafficking within Airway Epithelial Cells Using High-Resolution Automated Quantitative Confocal Microscopy |
| title_short | Characterisation of <i>Aspergillus fumigatus</i> Endocytic Trafficking within Airway Epithelial Cells Using High-Resolution Automated Quantitative Confocal Microscopy |
| title_sort | characterisation of i aspergillus fumigatus i endocytic trafficking within airway epithelial cells using high resolution automated quantitative confocal microscopy |
| topic | <i>Aspergillus fumigatus</i> airway epithelial cells phagocytosis |
| url | https://www.mdpi.com/2309-608X/7/6/454 |
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