Inactivating conditions of therapeutic mycobacteriophages

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 re..., , # Inactivating conditions of therapeutic mycobacteriophages

[https://doi.org/10.5061/dryad.t1g1jwt8h](https://doi.org/10.5061/dryad.t1g1jwt8h)

To measure phage sensitivities, phages at titers of 10^9 – 10^10 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...,

目前亟需开发新型疗法以治疗耐药性非结核分枝杆菌（nontuberculous mycobacteria, NTM）疾病。噬菌体（bacteriophages，简称phages）是一类可感染并裂解细菌的病毒，目前正被积极探索作为治疗分枝杆菌疾病的替代方案。多项针对耐药性NTM感染的噬菌体治疗同情用药案例显示出良好的治疗效果。为进一步推动噬菌体疗法的发展，需识别并规避在噬菌体生产、储存、制剂配制或治疗过程中会对噬菌体活性产生负面影响的环境条件。反之，在部分临床前噬菌体疗法实验中，则需要灭活游离的噬菌体。本研究针对3株来自耐药NTM同情用药治疗案例的分枝杆菌噬菌体——BPsΔ33HTH-HRM10、Muddy与ZoeJΔ4,5，探究其对温度、酸性pH值、去污剂、黏液及噬菌体灭活缓冲液等多种条件的敏感性。多种条件可导致噬菌体活性出现显著且快速的下降……
# 治疗用分枝杆菌噬菌体的灭活条件
https://doi.org/10.5061/dryad.t1g1jwt8h

为测定噬菌体的敏感性，本研究将滴度为10^9–10^10 噬菌斑形成单位/毫升（plaque-forming unit per ml, PFU/ml）的噬菌体置于分枝杆菌噬菌体（mycobacteriophage, MP）缓冲液中，在室温（22℃）条件下暴露于不同环境（另有说明的情况除外）。在指定时间点，采用双层琼脂平板噬菌斑测定法，以耻垢分枝杆菌（Mycobacterium smegmatis）为宿主菌，通过噬菌斑计数法测定噬菌体滴度（PFU/ml）。与同期未处理的噬菌体对照组相比，导致噬菌体滴度显著下降的条件可通过对数变化值计算得出：log PFUchallenge – log PFUcontrol。重复实验得到的对数变化数据已整理于论文的表1中。

Fig1ABCv2.csv：该逗号分隔值（Comma-Separated Values, CSV）文件记录了3株噬菌体（BPsΔ33HTH-HRM10、Muddy与ZoeJΔ4,5）经不同温度、不同pH值的PIB缓冲液等条件处理后的噬菌斑形成单位浓度（pfu/ml）及其以10为底的对数值[log(pfu/ml)]原始数据……