A depolymerase producing lytic bacteriophage (KPO1K2) (MTCC 5831), already characterized in our laboratory was used in the present study [14]

A depolymerase producing lytic bacteriophage (KPO1K2) (MTCC 5831), already characterized in our laboratory was used in the present study [14]. compared. Also, an attempt to compare the efficacy of PNZ5 free phage and liposome entrapped phage, alone or in conjunction with amikacin in eradicating mature biofilm was made. The entrapment of phage in liposome provided 100% protection to phage from neutralizing antibody. PNZ5 On the contrary un-entrapped phage got neutralized within 3 h of its interaction with antibody. Compared to the inability of free phage to enter macrophages, the liposome were able to deliver entrapped phage inside macrophages and cause 94.6% killing of intracellular as well as by overcoming the majority of the hurdles related to clinical use of phage. Introduction is an important pathogen that accounts for up to 8% of nosocomial infections in the Western world. It is placed among the eight most infectious agents in hospitals [1]. Its incidence PNZ5 is often linked to use of artificial devices, indicating that bacterial adhesion and biofilm formation are important virulence attributes in the establishment of infection. Lytic bacteriophage with antibacterial property has been recommended as an adjunct therapy to antibiotics as it offers a number of advantages over the existing chemical agents. However, production of neutralizing antibodies, emergence of phage-resistant bacterial strains and restricted intracellular entry of phage are considered as major hurdles in the clinical use of phage [2C5]. The production of anti-phage PNZ5 antibodies has been reported following phage therapy [6,7]. These antibodies have the ability to inactivate the phage due to their neutralizing nature. However, the actual impact of these antibodies on the efficacy of repeated phage dosing is unclear. Previous researchers have suggested that bacteriophages can be used to treat bacterial infections as long as bacteria remain accessible. However, it has also been suggested that phage therapy is not suitable for intracellular bacteria because of their inability to enter into myeloid cells. was initially considered as an extracellular pathogen but now it is clear that it is also capable of intracellular survival in a variety of cells including phagocytic cells (macrophages) and epithelial cells [8,9,10,11]. Survival of within epithelial cells PNZ5 may serve as a critical reservoir from where reinfection of the host can take place. Most of the antibiotics show poor penetration within eukaryotic cells, leading to persistence of Rabbit Polyclonal to ASC pathogens. Recently, we have reported that entrapment of phage in liposome could be considered as a powerful tool against induced respiratory tract infection in mice as it resulted in mounting a highly effective therapeutic and prophylactic response [12]. The present study was designed to ascertain the reason for the higher efficacy of liposome entrapped phage as compared to free phage. In the present study, liposome were used as a delivery vehicle for the already characterized lytic phage KPO1K2 and this liposome entrapped phage alone or in conjunction with antibiotic was evaluated for its ability to eradicate mature biofilm of persisting within murine macrophages was also attempted using experiments. Materials and Methods Ethics Statement The experimental protocols were approved by the Institutional Animal Ethics Committee of Panjab University, Chandigarh, India (Approval ID: IAEC/156). Animal experimentation was performed in accordance with the guidelines of Committee for the Purpose of Control and Supervision of Experiments on Animals (CPCSEA), Government of India. All efforts were made to minimize the suffering of animals. Bacterial strain, bacteriophage and antibiotic B5055 (MTCC 5832) obtained originally from Dr M. Trautman, Department of Medical Microbiology and Hygiene, University of Ulm, Ulm, Germany and maintained in the laboratory was used in the present study. This strain expresses O1 and K2 antigens, which makes it the most commonly encountered serotype in clinical situations. All experiments were performed in nutrient broth medium. Growth media and antibiotic were purchased from HiMedia Laboratories, Mumbai, India. Stock solution of.