Efficacy of Bacteriophage Therapy in a Model ofBurkholderia cenocepaciaPulmonary Infection

Abstract

The therapeutic potential of bacteriophage (phage) in a mouse model of acute B. cenocepacia pulmonary infection was assessed. Phage were administered by either intranasal (i.n.) inhalation or intraperitoneal (i.p.) injection. Bacterial density, macrophage inflammatory protein-2 (MIP-2), and tumor necrosis factor-α (TNFα) levels were significantly reduced in lungs of mice treated with i.p. phage. No significant differences in lung bacterial density or MIP-2 levels were found between untreated mice and mice treated with i.n. phage, i.p. UV-inactivated phage, or i.p. λ phage controls. Mock-infected mice treated with phage showed no significant increase in lung MIP-2 or TNFα levels compared to mock-infected / mock-treated mice. We have demonstrated the efficacy of phage therapy in an acute B. cenocepacia lung infection model. Systemic administration of phage was more effective than inhalational administration, suggesting that circulating phage have better access to bacteria in lung compared to topical phage.

Cystic Fibrosis Therapy: A Community Ecology Perspective

“…Significantly, phages were absent from late stage biofilms in CF lungs, suggesting that biofilm formation may help bacteria escape this top-down control. Further characterization of the phage community and its impact on bacteria present in the CF lung can be expected to provide new insights into the top-down control of the infection, including the potential use of phages for novel therapeutic options (26).…”

Identification and characterization of ϕH111-1

“…4 Although the therapeutic use of temperate phages is generally discouraged, the limited availability of obligately lytic BCC phages has necessitated the use of confirmed, putative, or modified temperate phages for several in vivo efficacy studies. [6][7][8] The use of such phages against Burkholderia is arguably safer than it is against many other pathogens because virulence factors in this genus have not been discovered to be encoded by temperate phages. 2 One of the challenges of prophage identification is the differentiation of inducible prophages from defective prophage remnants, a distinction with both evolutionary and practical implications.…”

Genomes and Characterization of Phages Bcep22 and BcepIL02, Founders of a Novel Phage Type in Burkholderia cenocepacia

“…Phage was then dialyzed thoroughly against gelatin-free SM buffer and concentrated further using a 100-kDa molecular weight cutoff Amicon ultrafiltration unit (Millipore). Phage proteins were analyzed by SDS-PAGE by boiling purified phage for 5 min in Laemmli sample buffer (62.5 mM Tris-HCl [pH 6.8], 2% SDS, 10% glycerol, 5% ␤-mercaptoethanol, 0.001% bromophenol blue), and the equivalent of ϳ3 ϫ 10 10 PFU was loaded per lane on a 10% SDS-PAGE gel by standard methods (40). Coomassie-stained bands were excised and subjected to reduction using dithiothreitol and alkylation with iodoacetamide followed by tryptic digestion carried out robotically using a ProGest robotic device (Genomic Solutions, Ann Arbor, MI) suited for automated digestion protocols and temperature-controlled reactions.…”