Bacteriophages φMR299-2 and φNH-4 can eliminate Pseudomonas aeruginosa in the murine lung and on cystic fibrosis lung airway cells.
Bacteriophages φMR299-2 and φNH-4 can eliminate Pseudomonas aeruginosa in the murine lung and on cystic fibrosis lung airway cells.
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作者：
Alemayehu D;Casey PG;McAuliffe O;Guinane CM;Martin JG;Shanahan F;Coffey A;Ross RP;Hill C
铜绿假单胞菌是囊性纤维化（CF）患者肺部感染的常见原因。此外，假单胞菌的生物膜形成和抗生素耐药性是使抗生素治疗复杂化的主要问题。我们评估了使用噬菌体杀死生物膜和鼠肺中病原体的功效。我们从当地的污水处理厂分离和表征两种病毒，一种肌病毒（MNH-4）和一种足状病毒（MMR 299 -2）。两种抗生素均对铜绿假单胞菌的临床分离株具有活性。两种细菌一起杀死了所有9种测试的临床分离菌株，包括粘液和非粘液菌株。当作为生物膜在囊性纤维化支气管上皮细胞CFBE 410-细胞系上生长时，两种抗生素的等量混合物可有效杀死铜绿假单胞菌NH 57388 A（粘液）和铜绿假单胞菌MR 299（非粘液）菌株.噬菌体滴度在24小时内增加了近100倍，证实了噬菌体的复制。此外，噬菌体混合物在含有1 × 107至2 × 107铜绿假单胞菌的小鼠肺中也能有效杀死病原体。假单胞菌在6小时内从鼠肺中有效清除（减少至少3至4个对数单位的量级）。我们的研究证明了这两种噬菌体在杀死小鼠肺部临床假单胞菌分离株或作为肺部细胞系上的生物膜方面的功效，并支持了人们对使用噬菌体疗法控制和治疗CF患者多药耐药假单胞菌肺部感染日益增长的兴趣。鉴于抗生素耐药性的上升，需要非抗生素疗法来治疗感染。对于囊性纤维化患者的假单胞菌感染的治疗尤其如此。我们已经鉴定了两种细菌病毒（噬菌体），它们可以杀死生长在人类肺细胞上和肺部感染动物模型中的假单胞菌。使用噬菌体是特别合适的，因为杀伤剂可以在靶细胞上复制，产生噬菌体的新鲜拷贝。因此，在目标存在的情况下，杀伤剂倍增。通过使用两种噬菌体，我们可以降低在感染部位形成耐药菌落的风险。噬菌体治疗是一个令人兴奋的领域，这项研究代表了有效的感染模型的有效性的重要证明。
Pseudomonas aeruginosa is a common cause of infection in the lungs of patients with cystic fibrosis (CF). In addition, biofilm formation and antibiotic resistance of Pseudomonas are major problems that can complicate antibiotic therapy. We evaluated the efficacy of using bacteriophages to kill the pathogen in both biofilms and in the murine lung. We isolated and characterized two phages from a local wastewater treatment plant, a myovirus (ϕNH-4) and a podovirus (ϕMR299-2). Both phages were active against clinical isolates of P. aeruginosa. Together, the two phages killed all 9 clinical isolate strains tested, including both mucoid and nonmucoid strains. An equal mixture of the two phages was effective in killing P. aeruginosa NH57388A (mucoid) and P. aeruginosa MR299 (nonmucoid) strains when growing as a biofilm on a cystic fibrosis bronchial epithelial CFBE41o- cell line. Phage titers increased almost 100-fold over a 24-h period, confirming replication of the phage. Furthermore, the phage mix was also effective in killing the pathogen in murine lungs containing 1 × 107 to 2 × 107 P. aeruginosa. Pseudomonas was effectively cleared (reduced by a magnitude of at least 3 to 4 log units) from murine lungs in 6 h. Our study demonstrates the efficacy of these two phages in killing clinical Pseudomonas isolates in the murine lung or as a biofilm on a pulmonary cell line and supports the growing interest in using phage therapy for the control and treatment of multidrug-resistant Pseudomonas lung infections in CF patients. Given the rise in antibiotic resistance, nonantibiotic therapies are required for the treatment of infection. This is particularly true for the treatment of Pseudomonas infection in patients with cystic fibrosis. We have identified two bacterial viruses (bacteriophages) that can kill Pseudomonas growing on human lung cells and in an animal model of lung infection. The use of bacteriophages is particularly appropriate because the killing agent can replicate on the target cell, generating fresh copies of the bacteriophage. Thus, in the presence of a target, the killing agent multiplies. By using two bacteriophages we can reduce the risk of resistant colonies developing at the site of infection. Bacteriophage therapy is an exciting field, and this study represents an important demonstration of efficacy in validated infection models.