A novel phage carrying capsule depolymerase effectively relieves pneumonia caused by multidrug-resistant Klebsiella aerogenes

• Published in: Journal of biomedical science 2023-08, Vol.30 (1), p.1-75, Article 75
• Main Authors: Cui, Xiaohu, Du, Bing, Feng, Junxia, Feng, Yanling, Fan, Zheng, Chen, Jinfeng, Cui, Jinghua, Gan, Lin, Fu, Tongtong, Tian, Ziyan, Zhang, Rui, Yan, Chao, Zhao, Hanqing, Xu, Wenjian, Xu, Ziying, Yu, Zihui, Ding, Zanbo, Li, Zhoufei, Chen, Yujie, Xue, Guanhua, Yuan, Jing
• Format: Article
• Language: English
• Subjects: Adaptability; Aerogenes; Analysis; Antibiotic resistance; Antibiotics; Antimicrobial agents; Bacteria; Bacterial pneumonia; Bacteriophage therapy; Biofilms; Bioinformatics; Confocal microscopy; Depolymerase; Drug resistance in microorganisms; Gene sequencing; Genomes; Genomics; Health aspects; Klebsiella; Klebsiella aerogenes; Laboratory animals; Larvae; Maximum likelihood method; Microscopy; Multidrug resistance; Multidrug resistant organisms; Nosocomial infections; Phages; Phenols; Physical characteristics; Plaque assay; Pneumonia; Proteins; Scanning microscopy; Sewage; Side effects; Trachea; Transmission electron microscopy; Ventilator-associated pneumonia; Virulence; Virulence (Microbiology); Whole genome sequencing
• ISSN: 1423-0127; 1021-7770; 1423-0127
• DOI: 10.1186/s12929-023-00946-y

Background Klebsiella aerogenes can cause ventilator-associated pneumonia by forming biofilms, and it is frequently associated with multidrug resistance. Phages are good antibiotic alternatives with unique advantages. There has been a lack of phage therapeutic explorations, kinetic studies, and interaction mechanism research targeting K. aerogenes. Methods Plaque assay, transmission electron microscopy and whole-genome sequencing were used to determine the biology, morphology, and genomic characteristics of the phage. A mouse pneumonia model was constructed by intratracheal/endobronchial delivery of K. aerogenes to assess the therapeutic effect of phage in vivo. Bioinformatics analysis and a prokaryotic protein expression system were used to predict and identify a novel capsule depolymerase. Confocal laser scanning microscopy, Galleria mellonella larvae infection models and other experiments were performed to clarify the function of the capsule depolymerase. Results A novel lytic phage (pK4-26) was isolated from hospital sewage. It was typical of the Podoviridae family and exhibited serotype specificity, high lytic activity, and high environmental adaptability. The whole genome is 40,234 bp in length and contains 49 coding domain sequences. Genomic data show that the phage does not carry antibiotic resistance, virulence, or lysogenic genes. The phage effectively lysed K. aerogenes in vivo, reducing mortality and alleviating pneumonia without promoting obvious side effects. A novel phage-derived depolymerase was predicted and proven to be able to digest the capsule, remove biofilms, reduce bacterial virulence, and sensitize the bacteria to serum killing. Conclusions The phage pK4-26 is a good antibiotic alternative and can effectively relieve pneumonia caused by multidrug-resistant K. aerogenes. It carries a depolymerase that removes biofilms, reduces virulence, and improves intrinsic immune sensitivity. Keywords: Klebsiella aerogenes, Pneumonia, Bacteriophage therapy, Depolymerase, Biofilms