Inactivating conditions of therapeutic mycobacteriophages
Data files
Dec 18, 2025 version files 96.87 KB
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Fig1ABCv2.csv
40.14 KB
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README.md
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Supplemental_Fig_1.csv
427 B
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Table1_Dryad_Raw_Data_v2.csv
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Abstract
There is a need for new therapies to treat drug-resistant nontuberculous mycobacteria (NTM) disease. Bacteriophages (phages), which are viruses that infect and kill bacteria, are actively being explored as an alternative approach for treating mycobacterial diseases. Several compassionate-use cases of phage therapy for drug-resistant NTM infections exhibit favorable outcomes. To further the development of phage therapy, it is important to recognize and avoid conditions that negatively impact phage activity during phage production, storage, formulation, or treatment. Conversely, there is a need to inactivate free phages in certain preclinical phage therapy experiments. In this study, we investigated three mycobacteriophages, BPsΔ33HTH-HRM10, Muddy, and ZoeJΔ4,5 from compassionate-use NTM treatment cases for their sensitivity to a variety of conditions that included temperature, acid pH, detergents, mucus, and phage inactivating buffers. Several conditions resulted in dramatic and rapid reductions in the level of active phage, while others had no effect. We also observed different sensitivities between the phages. The results provide valuable information to support further investigation and development of these phages as therapeutics.
https://doi.org/10.5061/dryad.t1g1jwt8h
To measure phage sensitivities, phages at titers of 109 – 1010 PFU/ml in mycobacteriophage (MP) buffer were exposed to different conditions at room temperature (22° C), unless otherwise indicated. At specific time points, phages were quantified as PFU/ml using an agar overlay assay with Mycobacterium smegmatis. Conditions resulting in a significant reduction in titer compared to an untreated phage control at the same time point were determined by calculating the log change (log PFUchallenge – log PFUcontrol. Log change data from replicate experiments are presented in Table 1 in the manuscript.
Fig1ABCv2.csv: CSV file reporting raw data for plaque-forming units per ml (pfu/ml) and the log10 transformation of the plaque-forming units per ml [log(pfu/ml)] for phages BPs, Muddy, and ZoeJ challenged to various temperatures, PIB at various pH, and MP Buffer at various pH.
Fig1A: Temperature sensitivity
Fig1B: PIB + pH Sensitivity
Fig1C: MP Buffer + pH Sensitivity
Table1_Dryad_Raw_Data_v2.csv: CSV file reporting raw data for plaque-forming units per ml (pfu/ml) and the log10 transformation of the plaque-forming units per ml [log(pfu/ml)] for phages BPs, Muddy, and ZoeJ challenged to various therapeutic and storage conditions.
Data for the conditions of Temperature, PIB, and MP Buffer are located in the file for Fig1ABC.
Supplemental_Fig_1.csv: CSV file reporting raw data colony-forming units per ml (cfu/ml) and the log10 transformation of colony-forming units per ml [log(cfu/ml)] for Mycobacterium smegmatis sensitivity to phage inhibition buffer (PIB) at pH 3.
Supplemental Fig 1. Mycobacterium smegmatis is not sensitive to phage inhibition buffer (PIB) at pH 3.