title: Quality-controlled small-scale production of a well-defined bacteriophage cocktail for use in human clinical trials.
abstract: We describe the small-scale, laboratory-based, production and quality control of a cocktail, consisting of exclusively lytic bacteriophages, designed for the treatment of Pseudomonas aeruginosa and Staphylococcus aureus infections in burn wound patients. Based on successive selection rounds three bacteriophages were retained from an initial pool of 82 P. aeruginosa and 8 S. aureus bacteriophages, specific for prevalent P. aeruginosa and S. aureus strains in the Burn Centre of the Queen Astrid Military Hospital in Brussels, Belgium. This cocktail, consisting of P. aeruginosa phages 14/1 (Myoviridae) and PNM (Podoviridae) and S. aureus phage ISP (Myoviridae) was produced and purified of endotoxin. Quality control included Stability (shelf life), determination of pyrogenicity, sterility and cytotoxicity, confirmation of the absence of temperate bacteriophages and transmission electron microscopy-based confirmation of the presence of the expected virion morphologic particles as well as of their specific interaction with the target bacteria. Bacteriophage genome and proteome analysis confirmed the lytic nature of the bacteriophages, the absence of toxin-coding genes and showed that the selected phages 14/1, PNM and ISP are close relatives of respectively F8, phiKMV and phage G1. The bacteriophage cocktail is currently being evaluated in a pilot clinical study cleared by a leading Medical Ethical Committee.
authors.affiliation: Eliava Institute of Bacteriophage, Microbiology and Virology, Tbilisi, Georgia.
authors.forename: Maya
authors.initials: M
authors.lastname: Merabishvili
authors.forename: Jean-Paul
authors.initials: JP
authors.lastname: Pirnay
authors.forename: Gilbert
authors.initials: G
authors.lastname: Verbeken
authors.forename: Nina
authors.initials: N
authors.lastname: Chanishvili
authors.forename: Marina
authors.initials: M
authors.lastname: Tediashvili
authors.forename: Nino
authors.initials: N
authors.lastname: Lashkhi
authors.forename: Thea
authors.initials: T
authors.lastname: Glonti
authors.forename: Victor
authors.initials: V
authors.lastname: Krylov
authors.forename: Jan
authors.initials: J
authors.lastname: Mast
authors.forename: Luc
authors.initials: L
authors.lastname: Van Parys
authors.forename: Rob
authors.initials: R
authors.lastname: Lavigne
authors.forename: Guido
authors.initials: G
authors.lastname: Volckaert
authors.forename: Wesley
authors.initials: W
authors.lastname: Mattheus
authors.forename: Gunther
authors.initials: G
authors.lastname: Verween
authors.forename: Peter
authors.initials: P
authors.lastname: De Corte
authors.forename: Thomas
authors.initials: T
authors.lastname: Rose
authors.forename: Serge
authors.initials: S
authors.lastname: Jennes
authors.forename: Martin
authors.initials: M
authors.lastname: Zizi
authors.forename: Daniel
authors.initials: D
authors.lastname: De Vos
authors.forename: Mario
authors.initials: M
authors.lastname: Vaneechoutte
mesh_terms: D001435:Bacteriophages / Q000235:genetics* / Q000378:metabolism* / Q000648:ultrastructure; D002056:Burns / Q000150:complications / Q000382:microbiology; D002986:Clinical Trials as Topic; D016679:Genome, Viral; D006801:Humans; D020543:Proteome / Q000032:analysis / Q000235:genetics; D011552:Pseudomonas Infections / Q000628:therapy*; D011550:Pseudomonas aeruginosa / Q000235:genetics / Q000821:virology; D013203:Staphylococcal Infections / Q000628:therapy*; D013211:Staphylococcus aureus / Q000235:genetics / Q000821:virology; D014946:Wound Infection / Q000382:microbiology / Q000628:therapy
chemical_list: D020543:Proteome