title: Cheating by type 3 secretion system-negative Pseudomonas aeruginosa during pulmonary infection.
abstract: The opportunistic pathogen Pseudomonas aeruginosa expresses a type 3 secretion system (T3SS) strongly associated with bacterial virulence in murine models and human patients. T3SS effectors target host innate immune mechanisms, and T3SS-defective mutants are cleared more efficiently than T3SS-positive bacteria by an immunocompetent host. Nonetheless, T3SS-negative isolates are recovered from many patients with documented P. aeruginosa infections, leading us to test whether T3SS-negative strains could have a selective advantage during in vivo infection. Mice were infected with mixtures of T3SS-positive WT P. aeruginosa plus isogenic T3SS-OFF or constitutively T3SS-ON mutants. Relative fitness of bacteria in this acute pneumonia model was reflected by the competitive index of mutants relative to WT. T3SS-OFF strains outcompeted WT PA103 in vivo, whereas a T3SS-ON mutant showed decreased fitness compared with WT. In vitro growth rates of WT and T3SS-OFF bacteria were determined under T3SS-inducing conditions and did not differ significantly. Increased fitness of T3SS-OFF bacteria was no longer observed at high ratios of T3SS-OFF to WT, a feature characteristic of bacterial cheaters. Cheating by T3SS-OFF bacteria occurred only when T3SS-positive bacteria expressed the phospholipase A2 effector Exotoxin U (ExoU). T3SS-OFF bacteria showed no fitness advantage in competition experiments carried out in immunodeficient MyD88-knockout mice or in neutrophil-depleted animals. Our findings indicate that T3SS-negative isolates benefit from the public good provided by ExoU-mediated killing of recruited innate immune cells. Whether this transient increase in fitness observed for T3SS-negative strains in mice contributes to the observed persistence of T3SS-negative isolates in humans is of ongoing interest.
authors.affiliation: Departments of Medicine and barbara.kazmierczak@yale.edu.
authors.forename: Kamila
authors.initials: K
authors.lastname: Czechowska
authors.affiliation: Departments of Medicine andGraduate Program in Molecular and Cell Biology, Quinnipiac University, Hamden, CT 06518.
authors.forename: Saria
authors.initials: S
authors.lastname: McKeithen-Mead
authors.affiliation: Departments of Medicine and.
authors.forename: Khatoun
authors.initials: K
authors.lastname: Al Moussawi
authors.affiliation: Departments of Medicine andMicrobial Pathogenesis, Yale University School of Medicine, New Haven, CT 06520; and barbara.kazmierczak@yale.edu.
authors.forename: Barbara I
authors.initials: BI
authors.lastname: Kazmierczak
mesh_terms: D000704:Analysis of Variance; D000818:Animals; D058947:Bacterial Secretion Systems / Q000276:immunology*; D060085:Coinfection / Q000276:immunology*; D016207:Cytokines / Q000276:immunology; D017931:DNA Primers / Q000235:genetics; D004797:Enzyme-Linked Immunosorbent Assay; D005098:Exotoxins / Q000378:metabolism; D005434:Flow Cytometry; D051379:Mice; D008856:Microscopy, Fluorescence; D010957:Plasmids / Q000235:genetics; D011014:Pneumonia / Q000276:immunology* / Q000382:microbiology; D011552:Pseudomonas Infections / Q000276:immunology*; D011550:Pseudomonas aeruginosa / Q000254:growth & development / Q000276:immunology*
chemical_list: D058947:Bacterial Secretion Systems; D016207:Cytokines; D017931:DNA Primers; D005098:Exotoxins