Data from: Bacteriophage infection and killing of intracellular Mycobacterium abscessus
Data files
Nov 27, 2023 version files 6.45 KB
Abstract
Mycobacterium abscessus is a nontuberculous mycobacterium (NTM) that contributes to the decline and death of patients with lung diseases such as cystic fibrosis and other muco-obstructive airway diseases. M. abscessus is challenging to treat due to its extensive antibiotic resistance and ability to survive inside mammalian cells. An alternative to antibiotics is the therapeutic use of bacteriophages (phages). There are recent cases of phage therapy being used to treat M. abscessus infections in people under compassionate-use conditions. However, little is known about the ability of phages to kill bacteria, such as M. abscessus, that reside in an intracellular environment. Here, we used M. abscessus strains and phages from recent phage therapy cases to determine if phages can enter mammalian cells and if they can infect and kill intracellular M. abscessus. Using fluorescence microscopy, we demonstrate phage uptake by macrophages and lung epithelial cells, and we further demonstrate phage infection of intracellular M. abscessus with fluorescent reporter phages. Transmission electron microscopy was additionally used to image phage infection of intracellular M. abscessus. Together, these findings provide the first visualizations of phage-M. abscessus interactions in an intracellular environment. Finally, we show that phage treatment can significantly reduce the intracellular burden of M. abscessus in a manner that depends on both the specific phage and mammalian cell type involved. These results demonstrate the potential to use phage therapy to treat intracellular bacteria, specifically M. abscessus, while also highlighting the importance of prescreening phage therapy candidates for activity in an intracellular environment.
README: Data from Bacteriophage Infection and Killing of Intracellular Mycobacterium abscessus
https://doi.org/10.5061/dryad.t1g1jwt8h
Figure 2ABCD
Fig2ABCD.csv: CSV file reporting raw data for bacterial colony forming unit (CFU) counts from time points between day 0 and day 5 post-infection for Figure 2ABCD. Raw values were transformed log10 for publication.
Figure 4BCD
Fig4BCD.csv: CSV file reporting raw data for quantification of phage puncta per mammalian cell in individual fields of view for Figure 4BCD. Values represent the percent of mammalian cells with intracellular phage.
Figure 5BCD
Fig5BDC.csv: CSV file reporting raw data for quantification of bacteria expressing mCherry as an indicator of phage infection for Figure 5BCD. Values represent percent reporter phage infection of intracellular M. abscessus.
Figure 7ABC
Fig7ABC.csv: CSV file reporting raw data for bacterial colony forming unit (CFU) counts from time points at day 0 and day 2 post-infection for Figure 7ABC. Raw values were transformed log10 for publication.
Figure S1
FigS1.csv: CSV file reporting raw data for plaque forming unit (PFU) counts +/- SYBR Gold staining of phage for Figure S1. Raw values were transformed log10 for publication.
Figure S6B
FigS6B.csv: CSV file raw data for bacterial colony forming unit (CFU) counts from bacterial samples treated +/- Triton +/- Phage Inactivation Buffer (PIB) for Figure S6B. Raw values were transformed log10 for publication.