Single Cell Analysis of Drug Susceptibility of <i>Mycobacterium abscessus</i> during Macrophage Infection
<i>Mycobacterium abscessus</i> is an emerging health risk to immunocompromised individuals and to people with pre-existing pulmonary conditions. As <i>M. abscessus</i> possesses multiple mechanisms of drug resistance, treatments of <i>M. abscessus</i> are of poor...
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MDPI AG
2020-10-01
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| Series: | Antibiotics |
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| Online Access: | https://www.mdpi.com/2079-6382/9/10/711 |
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| author | Joanna Brzostek Amierah Fatin Wen Hui Chua Hui Yi Tan Thomas Dick Nicholas R. J. Gascoigne |
| author_facet | Joanna Brzostek Amierah Fatin Wen Hui Chua Hui Yi Tan Thomas Dick Nicholas R. J. Gascoigne |
| author_sort | Joanna Brzostek |
| collection | DOAJ |
| description | <i>Mycobacterium abscessus</i> is an emerging health risk to immunocompromised individuals and to people with pre-existing pulmonary conditions. As <i>M. abscessus</i> possesses multiple mechanisms of drug resistance, treatments of <i>M. abscessus</i> are of poor efficacy. Therefore, there is an urgent need for new therapeutic strategies targeting <i>M. abscessus</i>. We describe an experimental system for screening of compounds for their antimicrobial activity against intracellular <i>M. abscessus</i> using flow cytometry and imaging flow cytometry. The assay allows simultaneous analysis of multiple parameters, such as proportion of infected host cells, bacterial load per host cell from the infected population, and host cell viability. We verified the suitability of this method using two antibiotics with known activity against <i>M. abscessus</i>: clarithromycin and amikacin. Our analysis revealed a high degree of infection heterogeneity, which correlated with host cell size. A higher proportion of the larger host cells is infected with <i>M. abscessus</i> as compared to smaller host cells, and infected larger cells have higher intracellular bacterial burden than infected smaller cells. Clarithromycin treatment has a more pronounced effect on smaller host cells than on bigger host cells, suggesting that heterogeneity within the host cell population has an effect on antibiotic susceptibility of intracellular bacteria. |
| first_indexed | 2024-03-10T15:32:50Z |
| format | Article |
| id | doaj.art-c66e6409d24d435f8fcec855ae7fd42e |
| institution | Directory Open Access Journal |
| issn | 2079-6382 |
| language | English |
| last_indexed | 2024-03-10T15:32:50Z |
| publishDate | 2020-10-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Antibiotics |
| spelling | doaj.art-c66e6409d24d435f8fcec855ae7fd42e2023-11-20T17:30:37ZengMDPI AGAntibiotics2079-63822020-10-0191071110.3390/antibiotics9100711Single Cell Analysis of Drug Susceptibility of <i>Mycobacterium abscessus</i> during Macrophage InfectionJoanna Brzostek0Amierah Fatin1Wen Hui Chua2Hui Yi Tan3Thomas Dick4Nicholas R. J. Gascoigne5Department of Microbiology and Immunology, Yong Loo Lin School of Medcine, National University of Singapore, 5 Science Drive 2, Singapore 117545, SingaporeDepartment of Microbiology and Immunology, Yong Loo Lin School of Medcine, National University of Singapore, 5 Science Drive 2, Singapore 117545, SingaporeDepartment of Microbiology and Immunology, Yong Loo Lin School of Medcine, National University of Singapore, 5 Science Drive 2, Singapore 117545, SingaporeDepartment of Microbiology and Immunology, Yong Loo Lin School of Medcine, National University of Singapore, 5 Science Drive 2, Singapore 117545, SingaporeDepartment of Microbiology and Immunology, Yong Loo Lin School of Medcine, National University of Singapore, 5 Science Drive 2, Singapore 117545, SingaporeDepartment of Microbiology and Immunology, Yong Loo Lin School of Medcine, National University of Singapore, 5 Science Drive 2, Singapore 117545, Singapore<i>Mycobacterium abscessus</i> is an emerging health risk to immunocompromised individuals and to people with pre-existing pulmonary conditions. As <i>M. abscessus</i> possesses multiple mechanisms of drug resistance, treatments of <i>M. abscessus</i> are of poor efficacy. Therefore, there is an urgent need for new therapeutic strategies targeting <i>M. abscessus</i>. We describe an experimental system for screening of compounds for their antimicrobial activity against intracellular <i>M. abscessus</i> using flow cytometry and imaging flow cytometry. The assay allows simultaneous analysis of multiple parameters, such as proportion of infected host cells, bacterial load per host cell from the infected population, and host cell viability. We verified the suitability of this method using two antibiotics with known activity against <i>M. abscessus</i>: clarithromycin and amikacin. Our analysis revealed a high degree of infection heterogeneity, which correlated with host cell size. A higher proportion of the larger host cells is infected with <i>M. abscessus</i> as compared to smaller host cells, and infected larger cells have higher intracellular bacterial burden than infected smaller cells. Clarithromycin treatment has a more pronounced effect on smaller host cells than on bigger host cells, suggesting that heterogeneity within the host cell population has an effect on antibiotic susceptibility of intracellular bacteria.https://www.mdpi.com/2079-6382/9/10/711<i>Mycobacterium abscessus</i>drug screensingle cell analysis |
| spellingShingle | Joanna Brzostek Amierah Fatin Wen Hui Chua Hui Yi Tan Thomas Dick Nicholas R. J. Gascoigne Single Cell Analysis of Drug Susceptibility of <i>Mycobacterium abscessus</i> during Macrophage Infection Antibiotics <i>Mycobacterium abscessus</i> drug screen single cell analysis |
| title | Single Cell Analysis of Drug Susceptibility of <i>Mycobacterium abscessus</i> during Macrophage Infection |
| title_full | Single Cell Analysis of Drug Susceptibility of <i>Mycobacterium abscessus</i> during Macrophage Infection |
| title_fullStr | Single Cell Analysis of Drug Susceptibility of <i>Mycobacterium abscessus</i> during Macrophage Infection |
| title_full_unstemmed | Single Cell Analysis of Drug Susceptibility of <i>Mycobacterium abscessus</i> during Macrophage Infection |
| title_short | Single Cell Analysis of Drug Susceptibility of <i>Mycobacterium abscessus</i> during Macrophage Infection |
| title_sort | single cell analysis of drug susceptibility of i mycobacterium abscessus i during macrophage infection |
| topic | <i>Mycobacterium abscessus</i> drug screen single cell analysis |
| url | https://www.mdpi.com/2079-6382/9/10/711 |
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