Development of a Bacteriophage Cocktail to Constrain the Emergence of Phage-Resistant Pseudomonas aeruginosa
Development of a Bacteriophage Cocktail to Constrain the Emergence of Phage-Resistant Pseudomonas aeruginosa
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开发噬菌体混合物来抑制噬菌体抗性铜绿假单胞菌的出现。
DOI：
10.3389/fmicb.2020.00327
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发表时间：
2020-03-04
影响因子：
5.2
通讯作者：
Le, Shuai
中科院分区：
文献类型：
--
作者：
Yang, Yuhui;Shen, Wei;Le, Shuai
随着多重耐药和广泛耐药细菌病原体的出现，噬菌体治疗和其他替代或额外的治疗方式正在重新受到关注。开发有效的噬菌体疗法的主要障碍之一是细菌宿主中噬菌体抗性的进化。当铜绿假单胞菌感染以O-抗原为受体的噬菌体时，噬菌体抗性通常通过改变或丧失O-抗原结构来实现。在这项研究中，我们表明，dsRNA噬菌体phiYY使用核心脂多糖作为受体，因此有效地杀死O-抗原缺失突变体。此外，通过噬菌体训练，我们获得了PaoP 5-m1，其是双链DNA噬菌体PaoP 5的衍生物，其能够感染具有截短的O-抗原的突变体。然后，我们通过将phiYY和PaoP 5-m1与另外三种宽宿主范围的铜绿假单胞菌混合来产生混合物。噬菌体混合物对铜绿假单胞菌临床分离株的多种选择有效，并且在短期内限制了困扰单个噬菌体有效性的噬菌体抗性突变体的出现。对5-噬菌体混合物的抗性在几天后出现，并且需要wzy和migA两者的突变。因此，本研究提供了用于设计噬菌体混合物和噬菌体治疗的替代策略。
With the emergence of multidrug-resistant and extensively drug-resistant bacterial pathogens, phage therapy and other alternative or additional therapeutic modalities are receiving resurgent attention. One of the major obstacles in developing effective phage therapies is the evolution of phage resistance in the bacterial host. When Pseudomonas aeruginosa was infected with a phage that uses O-antigen as receptor, phage resistances typically achieved through changing or loss of O-antigen structure. In this study, we showed that dsRNA phage phiYY uses core lipopolysaccharide as receptor and therefore efficiently kills the O-antigen deletion mutants. Furthermore, by phage training, we obtained PaoP5-m1, a derivative of dsDNA phage PaoP5, which is able to infect mutants with truncated O-antigen. We then generated a cocktail by mixing phiYY and PaoP5-m1 with additional three wide host range P. aeruginosa phages. The phage cocktail was effective against a diverse selection of clinical isolates of P. aeruginosa, and in the short-term constrained the appearance of the phage-resistant mutants that had beleaguered the effectiveness of single phage. Resistance to the 5-phage cocktail emerges after several days, and requires mutations in both wzy and migA Thus, this study provides an alternative strategy for designing phage cocktail and phage therapy.