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New Pseudomonas infections drive Pf phage transmission in CF airways
Julie D. Pourtois, Naomi L. Haddock, Aditi Gupta, Arya Khosravi, Hunter A. Martinez, Amelia K. Schmidt, Prema S. Prakash, Ronit Jain, Piper Fleming, Tony H. Chang, Carlos Milla, Patrick R. Secor, Giulio A. De Leo, Paul L. Bollyky, Elizabeth B. Burgener
Julie D. Pourtois, Naomi L. Haddock, Aditi Gupta, Arya Khosravi, Hunter A. Martinez, Amelia K. Schmidt, Prema S. Prakash, Ronit Jain, Piper Fleming, Tony H. Chang, Carlos Milla, Patrick R. Secor, Giulio A. De Leo, Paul L. Bollyky, Elizabeth B. Burgener
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Research Article Infectious disease Microbiology

New Pseudomonas infections drive Pf phage transmission in CF airways

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Abstract

Pf bacteriophages, lysogenic viruses that infect Pseudomonas aeruginosa (Pa), are implicated in the pathogenesis of chronic Pa infections; phage-infected (Pf+) strains are known to predominate in people with cystic fibrosis (pwCF) who are older and have more severe disease. However, the transmission patterns of Pf underlying the progressive dominance of Pf+ strains are unclear. In particular, it is unknown whether phage transmission commonly occurs horizontally between bacteria via viral particles within the airway or whether Pf+ bacteria are mostly acquired via de novo Pseudomonas infections. Here, we studied Pa genomic sequences from 3 patient cohorts totaling 662 clinical isolates from 105 pwCF. We identified Pf+ isolates and analyzed transmission patterns of Pf within patients between genetically similar groups of bacteria called “clone types.” We found that Pf was predominantly passed down vertically within Pa clone types and rarely via horizontal transfer between clone types within the airway. Conversely, we found extensive evidence of Pa de novo infection by a new, genetically distinct Pf+ Pa. Finally, we observed that clinical isolates showed reduced activity of type IV pili and reduced susceptibility to Pf in vitro. These results cast light on the transmission of virulence-associated phages in the clinical setting.

Authors

Julie D. Pourtois, Naomi L. Haddock, Aditi Gupta, Arya Khosravi, Hunter A. Martinez, Amelia K. Schmidt, Prema S. Prakash, Ronit Jain, Piper Fleming, Tony H. Chang, Carlos Milla, Patrick R. Secor, Giulio A. De Leo, Paul L. Bollyky, Elizabeth B. Burgener

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Figure 5

Clinical isolates do not twitch and are less susceptible to Pf than PAO1.

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Clinical isolates do not twitch and are less susceptible to Pf than PAO1...
Clinical isolates were selected from patients who were infected by both Pf– and Pf+ isolates and that showed sufficient growth in vitro. (A) Positive (PAO1) and negative (PAO1ΔPilA) controls for twitching assays, showing motility of bacteria over agar after 24 hours. PAO1ΔPilA mutants do not have functional type IV pili and cannot twitch, resulting in a small stained area. (B) Twitching area for PAO1, PAO1ΔPilA, and Pf– and Pf+ clinical isolates, with 4 replicates per isolate (mixed-effects model with isolate as random variable). ***P < 0.001 by t test. A lower twitching area indicates a lower twitching ability. (C) Plaque assays for Pf4 on PAO1ΔPf4ΔPf6 with negative control, on CPA0056 (Pf–), and on CPA0053ΔPf, which are representative of results for all clinical isolates. Plaque assays for other Pf+ and Pf– clinical isolates tested are shown in Supplemental Figure 5, for a total of 12 clinical isolates.

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