Therapeutic bacteriophage compositions
A phage and composition technology, applied in the field of phage composition, can solve the problems of MRSA infection that is difficult to treat with conventional antibiotics
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Embodiment 1
[0097] Phage group assembly (assembly)
[0098] A composition (group) of four phages Sa87, J-Sa36, J-Sa37 and Sa83, which together have broad activity against a group of recent multiple S. aureus clinical isolates (unpublished data), was used in animal research. In an exemplary method, phage lysates were prepared using production hosts SPS1216 and SPS1226, which do not release endogenous prophage during the production cycle. Grow the culture to OD in the bioreactor prior to phage addition 600 is 0.2. Continue to incubate at 37°C and read absorbance at least every 60 minutes. The culture is harvested after bacterial lysis and impurities are separated from the phage by several filtration steps followed by a chromatography step that reduces debris such as host cell proteins and host cell DNA. After purification, use a spin column ( Ultra 15-100kDa, Merck Millipore, Darmstadt, Germany) additionally concentrated the lysate such that the final phage titer was ≥1×10 11 PFU / m...
Embodiment 2
[0100] bacterial strain
[0101] Inoculate frozen stocks of Staphylococcus aureus UNT109-3 and UNT144-3 (obtained from the UNT culture collection center) into Trypticase TM Soy Broth (BBL TM Laboratories)+5% defibrinated sheep blood (TSBb) and incubated at 37°C for 18 hours (shaking). After 18 hours, the culture was diluted 1:10 into fresh TSBb and incubated for a further 5 hours before being diluted in fresh TSB for inoculation into mice.
Embodiment 3
[0103] Evaluation of Staphylococcus aureus virulence in a mouse model of pneumonia
[0104]The sensitivity of two S. aureus clinical isolates UNT109-3 (NRS234, native valve endocarditis) and UNT144-3 that have been used in animal models of infection was assessed relative to a single phage and a panel of 4 phages. In some embodiments, both strains are fully susceptible to the group as well as to individual phages Sa87, J-Sa36 and Sa83. Phage Sa87, J-Sa36 and Sa83 showed improved efficacy against UNT109-3 compared to phage J-Sa37. The virulence of both strains was then assessed in a murine lung infection model. Female Hsd:ICR(CD1) mice (Harlan Laboratories, Houston, TX) were administered 150 and 100 mg / kg cyclophosphamide on days 4 and 1 before infection to reduce their neutropenia. Groups of 5 mice were then anesthetized by intraperitoneal (IP) injection of 0.15 mL of a mixture of ketamine hydrochloride (100 mg / kg body weight) and xylazine (10 mg / kg body weight). Once anes...
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