Journal: Cells
Article Title: A Novel Co-Culture Model Reveals Enhanced CFTR Rescue in Primary Cystic Fibrosis Airway Epithelial Cultures with Persistent Pseudomonas aeruginosa Infection
doi: 10.3390/cells12222618
Figure Lengend Snippet: CFTR correctors caused a decrease in P. aeruginosa burden when co-cultured with HBE. ( A , B ) Primary CF HBE cultures (F508del/F508del) were treated with 58 µg/mL tobramycin 3 h after P. aeruginosa PAO1 infection (+ Pa ) or mock infection with media alone (− Pa ). Infection/mock infection was for 24 h. Correctors (Corr) VX-661 (10 µM) and VX-659 (1 µM) or VX-445 (3 µM) were added basolaterally at the time of infection. ( A ) Epithelial cultures remained intact 24 h after P. aeruginosa infection, as measured by resistance in Ussing chambers (4 donors, n = 2–4 per donor). ( B ) P. aeruginosa burden from apical washings from vehicle or corrector-rescued CF HBE cultures. P. aeruginosa burden was decreased in the presence of CFTR correctors. Data points represent the average for each HBE culture donor (4 donors, n = 5–6 per donor). ( C , D ) Direct treatment (no HBE) of P. aeruginosa with CFTR modulators and tobramycin. ( C ) P. aeruginosa growth in cell culture medium in the presence of vehicle (DMSO) or CFTR modulators individually and in combination. ( D ) Minimum inhibitory concentration (MIC) of P. aeruginosa to tobramycin in the absence or presence of CFTR modulators or vehicle control (DMSO). Statistical significance was determined by one-way ANOVA with Dunnet’s multiple comparison, * p < 0.05, ns = not significant.
Article Snippet: A CFTR corrector cocktail containing 2 correctors, either VX-659 (1 µM) or VX-445 (3 µM) plus VX-661 (10 µM) (Selleck Chemicals LLC, Houston, TX, USA), was applied to HBE cultures on the basolateral side chronically for 24 h. Potentiator VX-770 (1 µM) was added to the apical side of cultures for 30 min during Ussing chamber measurement or 30 min prior to lysing for Western blot or mRNA analysis, to mimic acute VX-770 treatment.
Techniques: Cell Culture, Infection, Concentration Assay, Comparison
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