Phage resistance formation and fitness costs of hypervirulent Klebsiella pneumoniae mediated by K2 capsule-specific phage and the corresponding mechanisms

doi:10.3389/fmicb.2023.1156292

科研成果详情

题名	Phage resistance formation and fitness costs of hypervirulent Klebsiella pneumoniae mediated by K2 capsule-specific phage and the corresponding mechanisms
作者	Tang, Miran1; Huang, Zeyu1; Zhang, Xiaodong1; Kong, Jingchun2; Zhou, Beibei1; Han, Yijia2; Zhang, Yi 2; Chen, Lijiang1; Zhou, Tieli1
发表日期	2023-07-19
发表期刊	Frontiers in microbiology 影响因子和分区
语种	英语
原始文献类型	Journal Article
关键词	bacteriophage capsular polysaccharides fitness cost hypervirulent Klebsiella pneumoniae lipopolysaccharides phage resistance polysaccharide depolymerase virulence
其他关键词	VIRULENCE
摘要	Phage is promising for the treatment of hypervirulent Klebsiella pneumoniae (hvKP) infections. Although phage resistance seems inevitable, we found that there still was optimization space in phage therapy for hvKP infection.}, The clinical isolate K. pneumoniae FK1979 was used to recover the lysis phage ΦFK1979 from hospital sewage. Phage-resistant bacteria were obtained on LB agar and used to isolate phages from sewage. The plaque assay, transmission electron microscopy (TEM), multiplicity of infection test, one-step growth curve assay, and genome analysis were performed to characterize the phages. Colony morphology, precipitation test and scanning electron microscope were used to characterize the bacteria. The absorption test, spot test and efficiency of plating (EOP) assay were used to identify the sensitivity of bacteria to phages. Whole genome sequencing (WGS) was used to identify gene mutations of phage-resistant bacteria. The gene expression levels were detected by RT-qPCR. Genes knockout and complementation of the mutant genes were performed. The change of capsules was detected by capsule quantification and TEM. The growth kinetics, serum resistance, biofilm formation, adhesion and invasion to A549 and RAW 264.7 cells, as well as G. mellonella and mice infection models, were used to evaluate the fitness and virulence of bacteria., {AbstractText=Here, we demonstrated that K2 capsule type sequence type 86 hvKP FK1979, one of the main pandemic lineages of hvKP with thick capsule, rapidly developed resistance to a K2-specific lysis phage ΦFK1979 which was well-studied in this work to possess polysaccharide depolymerase. The phage-resistant mutants showed a marked decrease in capsule expression. WGS revealed single nucleotide polymorphism (SNP) in genes encoding RfaH, galU, sugar glycosyltransferase, and polysaccharide deacetylase family protein in the mutants. RfaH and galU were further identified as being required for capsule production and phage sensitivity. Expressions of genes involved in the biosynthesis or regulation of capsule and/or lipopolysaccharide significantly decreased in the mutants. Despite the rapid and frequent development of phage resistance being a disadvantage, the attenuation of virulence and fitness in vitro and in vivo indicated that phage-resistant mutants of hvKP were more susceptible to the immunity system. Interestingly, the newly isolated phages targeting mutants changed significantly in their plaque and virus particle morphology. Their genomes were much larger than and significantly different from that of ΦFK1979. They possessed much more functional proteins and strikingly broader host spectrums than ΦFK1979. Our study suggests that K2-specific phage has the potential to function as an antivirulence agent, or a part of phage cocktails combined with phages targeting phage-resistant bacteria, against hvKP-relevant infections
资助项目	National Natural Science Foundation of China [82172328]; Key Laboratory of Clinical Laboratory Diagnosis and Translational Research of Zhejiang Province [2022E10022]
出版者	FRONTIERS MEDIA SA
ISSN	1664-302X
EISSN	1664-302X
卷号	14
DOI	10.3389/fmicb.2023.1156292
页数	18
WOS类目	Microbiology
WOS研究方向	Microbiology
WOS记录号	WOS:001041013300001
收录类别	PUBMED ; SCIE ; SCOPUS
URL	查看原文
PubMed ID	37538841
SCOPUSEID	2-s2.0-85166547503
通讯作者地址	[Chen, Lijiang]Department of Clinical Laboratory,The First Affiliated Hospital of Wenzhou Medical University,Key Laboratory of Clinical Laboratory Diagnosis and Translational Research of Zhejiang Province,The First Affiliated Hospital of Wenzhou Medical University,Zhejiang,Wenzhou,China ; [Zhou, Tieli]Department of Clinical Laboratory,The First Affiliated Hospital of Wenzhou Medical University,Key Laboratory of Clinical Laboratory Diagnosis and Translational Research of Zhejiang Province,The First Affiliated Hospital of Wenzhou Medical University,Zhejiang,Wenzhou,China
Scopus学科分类	Microbiology;Microbiology (medical)
TOP期刊	TOP期刊
引用统计
文献类型	期刊论文
条目标识符	https://kms.wmu.edu.cn/handle/3ETUA0LF/181820
专题	附属第一医院检验医学院（生命科学学院、生物学实验教学中心）第一临床医学院（信息与工程学院）、附属第一医院检验医学院（生命科学学院、生物学实验教学中心）_基础检验学系第一临床医学院（信息与工程学院）、附属第一医院_临床检验诊断学
通讯作者	Chen, Lijiang; Zhou, Tieli
作者单位	1.Department of Clinical Laboratory,The First Affiliated Hospital of Wenzhou Medical University,Key Laboratory of Clinical Laboratory Diagnosis and Translational Research of Zhejiang Province,The First Affiliated Hospital of Wenzhou Medical University,Zhejiang,Wenzhou,China; 2.Department of Medical Lab Science,School of Laboratory Medicine and Life Science,Wenzhou Medical University,Zhejiang,Wenzhou,China
第一作者单位	附属第一医院; 临床检验诊断学; 第一临床医学院（信息与工程学院）、附属第一医院
通讯作者单位	附属第一医院; 临床检验诊断学; 第一临床医学院（信息与工程学院）、附属第一医院
第一作者的第一单位	附属第一医院
推荐引用方式 GB/T 7714	Tang, Miran,Huang, Zeyu,Zhang, Xiaodong,et al. Phage resistance formation and fitness costs of hypervirulent Klebsiella pneumoniae mediated by K2 capsule-specific phage and the corresponding mechanisms[J]. Frontiers in microbiology,2023,14.
APA	Tang, Miran., Huang, Zeyu., Zhang, Xiaodong., Kong, Jingchun., Zhou, Beibei., ... & Zhou, Tieli. (2023). Phage resistance formation and fitness costs of hypervirulent Klebsiella pneumoniae mediated by K2 capsule-specific phage and the corresponding mechanisms. Frontiers in microbiology, 14.
MLA	Tang, Miran,et al."Phage resistance formation and fitness costs of hypervirulent Klebsiella pneumoniae mediated by K2 capsule-specific phage and the corresponding mechanisms".Frontiers in microbiology 14(2023).