Kongsomboonchoke Pattida, Mongkolkarvin Panupon, Khunti Patiphan, Vijitphichiankul Jarukit, Nonejuie Poochit, Thiennimitr Parameth, Chaikeeratisak Vorrapon
Biotechnology program, Faculty of Science, Chulalongkorn University, Bangkok, Thailand.
Department of Microbiology, Faculty of Medicine, Chiang Mai University, Chiang Mai, Thailand.
Sci Rep. 2025 Apr 14;15(1):12832. doi: 10.1038/s41598-025-96561-y.
Urinary tract infections are commonly caused by uropathogenic Escherichia coli (UPEC). Due to the emergence of multidrug-resistant UPEC, rendering antibiotic treatment ineffective, phage combination-based therapy has been proposed as a potential alternative. Here, we present a formulation of a genetically diverse phage-derived cocktail that is rapidly customized for UPEC using E. coli UTI89 as a model strain. Through our rapid selection and combination of four phages against UPEC strain UTI89 (SR01, SR02, SR04, and Zappy) from our library, the combination of two lytic phages, SR02 and SR04, exhibits the strongest suppression of bacterial growth for at least 16 h, with no emergence of phage resistance observed in vitro. Phage SR02 undergoes subcellular activity for 25 min, producing approximately 106 progeny particles per cell, while SR04 completes its replication cycle in 20 min, generating around 564 progeny particles per cell. These two novel phages are genetically diverse, and their cocktail exhibited potent suppression of bacterial growth, independent of multiplicities of infection (MOIs), significantly reducing the viable bacterial counts after treatment in vitro. The phage cocktail has low immunogenicity and does not induce any proinflammatory gene responses in human bladder uroepithelial cells. Moreover, the cocktail effectively eradicates the invading UPEC strain UTI89 in the uroepithelial cells at a comparable level to that of phage SR04 alone, likely releasing some immunostimulatory agents that, in turn, trigger upregulation of MIP-3 and IL-8 genes. Altogether, this study offers an alternative pipeline for rapidly formulating genetically diverse phage-derived cocktails, which is specifically customized for targeted bacteria.
尿路感染通常由尿路致病性大肠杆菌（UPEC）引起。由于多重耐药性UPEC的出现，使抗生素治疗无效，基于噬菌体组合的疗法已被提出作为一种潜在的替代方法。在此，我们展示了一种基因多样化的噬菌体衍生鸡尾酒制剂，它以大肠杆菌UTI89作为模型菌株，可快速针对UPEC进行定制。通过从我们的文库中快速筛选和组合针对UPEC菌株UTI89的四种噬菌体（SR01、SR02、SR04和Zappy），两种裂解性噬菌体SR02和SR04的组合对细菌生长的抑制作用最强，至少持续16小时，且在体外未观察到噬菌体抗性的出现。噬菌体SR02的亚细胞活性持续25分钟，每个细胞产生约106个子代颗粒，而SR04在20分钟内完成其复制周期，每个细胞产生约564个子代颗粒。这两种新型噬菌体在基因上是多样化的，它们的鸡尾酒制剂对细菌生长表现出强大的抑制作用，与感染复数（MOI）无关，在体外治疗后显著降低了活菌数量。该噬菌体鸡尾酒制剂具有低免疫原性，不会在人膀胱尿路上皮细胞中诱导任何促炎基因反应。此外，该鸡尾酒制剂能有效根除尿路上皮细胞中入侵的UPEC菌株UTI89，其效果与单独使用噬菌体SR04相当，可能释放了一些免疫刺激剂，进而触发MIP - 3和IL - 8基因的上调。总之，本研究提供了一种快速配制基因多样化的噬菌体衍生鸡尾酒制剂的替代途径，该制剂是专门针对目标细菌定制的。
