Mitigation of evolved bacterial resistance to phage therapy

Wound infections are the main cause of sepsis in patients with burns and increase burn-related morbidity and mortality. Bacteriophages, natural bacterial viruses, are being considered as an alternative therapy to treat infections caused by multidrug-resistant bacteria. We aimed to compare the efficacy and tolerability of a cocktail of lytic anti-Pseudomonas aeruginosa bacteriophages with standard of care for patients with burns.

Significance Bacteriophages (“phages,” viruses that infect bacteria) are an important source of selection for bacterial populations. Phages use various structures to infect bacterial cells, and bacteria often evolve phage resistance by losing or modifying these structures. We examine a phage that uses two structures that also provide Escherichia coli cells with antibiotic resistance. We show that phage selection can result in bacteria evolving phage resistance by losing or modifying the structures. When phage resistance evolves, the bacteria sometimes also show increased antibiotic sensitivity. This result indicates an evolutionary trade-off between phage resistance and antibiotic resistance. However, we also discovered bacterial mutations that avoid the trade-off. We discuss the potential use of phage selection and evolutionary trade-offs in treating bacterial infections.

Anti-K 1 phages were more active in vitro and in vivo against an 018:K1:H7 ColV+ Escherichia coli strain, designated MW, than were other phages. A single intramuscular dose of one anti-K1 phage was more effective than multiple intramuscular does of tetracycline, ampicillin, chloramphenicol, or trimethoprim plus sulphafurazole in curing mice of a potentially lethal intramuscularly or intracerebrally induced infection of MW; it was at least as effective as multiple intramuscular doses of streptomycin. When MW and the phage were inoculated into different gastrocnemius muscles of the same mice, a rapid reduction in numbers of MW organisms occurred in the MW-inoculated muscle and in other tissues; the numbers of phage particles in the MW-inoculated muscle increased rapidly and greatly. MW failed to proliferate in the brains of intracerebrally infected mice that had been inoculated intramuscularly with the phage at the same time; many more phage particles were found in the brains of these mice than in other sites. The few phage-resistant mutants of MW found in the phage-treated mich were K1-; previous studies had shown such mutants to be of greatly reduced virulence. The phage administered intramuscularly 3-5 d before challenge with a potentially lethal intramuscularly induced infection of MW was protective, the protective effect varying between phage propagated on different bacterial strains.