Fig. 4: Combination treatment of PAO1-infected HPAEpiC after 24 h high-tidal stretch most effective. | Nature Communications

Fig. 4: Combination treatment of PAO1-infected HPAEpiC after 24 h high-tidal stretch most effective.

From: Adjunctive phage therapy improves antibiotic treatment of ventilator-associated-pneumonia with Pseudomonas aeruginosa

Fig. 4

a Experimental plan created in BioRender. Nouailles, G. (2025) https://BioRender.com/padj3bg. HPAEpiC are seeded on 6-well BioFlex plates (5 × 105 cells/well) and stretched for 24 h (18 % elongation; 20 stretch cycles/min) mimicking high tidal volume (HVt) mechanical ventilation, following PAO1 infection (MOI 1) and treatment with either meropenem (MIC = 1 µg/mL), the Pseudomonas-specific phage cocktail, or the combination of both with indicated concentrations. At indicated time points cell supernatants were analyzed for b CFUs, c inflammatory cytokines and d released LDH for cell death. b Logarithmic display (Y = log(CFU + 1)) of bacterial load at 4 hpi and 8 hpi. Levels of c IL-6, IL-8 and d LDH in collected supernatants, measured in duplicates according to manufacturer´s recommendations. LDH results reflecting cytotoxicity are expressed as percentage of total LDH release during cell lysis by 1% Triton X-100 for 4 h or 8 h, respectively. Data are shown as means of experiments with box plots depicting median, quartiles, and range, as determined by two-way ANOVA with Tukey´s multiple comparisons test: *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001; n = 3 independent experiments, while each condition was run in duplicates per experiment. Color and symbol coding for groups: Control group: circles and shades of gray, Meropenem group: squares and shades of green, Phages group: triangles and shades of blue; Combination group: open circles and shades of orange. Dotted line reflects detection limit. Cells without any infection or treatment serves as medium control, infected cells (MOI 1) without any treatment serves as infection control. CFUs Colony-forming units, HPAEpiC human primary alveolar epithelial cells, hpi hours post infection, IL Interleukin, LDH Lactate dehydrogenase, MIC minimum inhibitory concentration. P-values were adjusted for multiple testing, b exact p-values can be found in Supplementary Tables 2–4. Source data are provided as a Source Data file.

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