在噬菌体与替加环素联合作用下抗碳青霉烯类肺炎克雷伯氏菌外膜囊泡的蛋白质组学分析。

IF 4.6 2区 医学 Q1 MICROBIOLOGY
Jing Mao, Xiaoyu Yang, Cheng Yan, Fan Wang, Rui Zheng
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引用次数: 0

摘要

背景:肺炎克雷伯菌是临床实践中最常见的病原体。广谱抗生素的广泛使用导致了目前耐碳青霉烯类抗生素肺炎克雷伯菌在全球范围内的传播,对抗菌治疗效果和公共卫生构成了重大威胁。外膜囊泡 (OMV) 已被确定为能够促进毒力基因和耐药基因转移的载体。然而,在抗生素和噬菌体治疗等外部压力下,OMVs 在耐碳青霉烯类肺炎双球菌中的作用仍不清楚:为了分离和纯化噬菌体和替加环素压力下的 OMVs,我们对携带质粒介导的 blaNDM-1 和 blaKPC-2 基因的肺炎克菌 0692 进行了密度梯度分离。采用双层板法分离出了 MJ1,它能有效裂解肺炎克菌 0692 细胞。利用透射电子显微镜(TEM)对分离出的噬菌体进行表征,并提取 OMV 组进行相关形态鉴定。通过双喹啉酸测定法(BCA)和蛋白质组学分析确定了各 OMV 组的蛋白质含量:结果:肺炎克氏菌 0692 在不同环境刺激下释放出 OMV,经 TEM 鉴定,这些 OMV 具有 OMV 的典型结构和粒径。与未处理组相比,噬菌体或替加环素单独处理可使肺炎克菌 0692 分泌的 OMVs 平均蛋白质浓度略有增加。然而,当噬菌体处理与替加环素联合使用时,OMVs 的平均蛋白质浓度比单独使用替加环素时显著降低。蛋白质组学研究表明,OMVs包被了许多功能性蛋白质,在噬菌体和替加环素的不同外部压力下,肺炎克菌0692衍生的OMVs所携带的蛋白质与未处理组相比有明显的上调或下调:这项研究证实了 OMVs 能够携带丰富的蛋白质,并强调了 OMV 相关蛋白质在细菌对噬菌体和替加环素的反应中的重要作用,是微生物耐药性研究的一项重要进展。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Proteomic analysis of carbapenem-resistant Klebsiella pneumoniae outer membrane vesicles under the action of phages combined with tigecycline.

Background: Klebsiella pneumoniae is the most commonly encountered pathogen in clinical practice. Widespread use of broad-spectrum antibiotics has led to the current global dissemination of carbapenem-resistant K. pneumoniae, which poses a significant threat to antibacterial treatment efficacy and public health. Outer membrane vesicles (OMVs) have been identified as carriers capable of facilitating the transfer of virulence and resistance genes. However, the role of OMVs in carbapenem-resistant K. pneumoniae under external pressures such as antibiotic and phage treatments remains unclear.

Methods: To isolate and purify OMVs under the pressure of phages and tigecycline, we subjected K. pneumoniae 0692 harboring plasmid-mediated blaNDM-1 and blaKPC-2 genes to density gradient separation. The double-layer plate method was used to isolate MJ1, which efficiently lysed K. pneumoniae 0692 cells. Transmission electron microscopy (TEM) was used to characterize the isolated phages and extract OMV groups for relevant morphological identification. Determination of protein content of each OMV group was conducted through bicinchoninic acid assay (BCA) and proteomic analysis.

Results: K. pneumoniae 0692 released OMVs in response to different environmental stimuli, which were characterized through TEM as having the typical structure and particle size of OMVs. Phage or tigecycline treatment alone resulted in a slight increase in the mean protein concentration of OMVs secreted by K. pneumoniae 0692 compared to that in the untreated group. However, when phage treatment was combined with tigecycline, there was a significant reduction in the average protein concentration of OMVs compared to tigecycline treatment alone. Proteomics showed that OMVs encapsulated numerous functional proteins and that under different external stresses of phages and tigecycline, the proteins carried by K. pneumoniae 0692-derived OMVs were significantly upregulated or downregulated compared with those in the untreated group.

Conclusions: This study confirmed the ability of OMVs to carry abundant proteins and highlighted the important role of OMV-associated proteins in bacterial responses to phages and tigecycline, representing an important advancement in microbial resistance research.

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来源期刊
CiteScore
8.60
自引率
0.00%
发文量
49
审稿时长
>12 weeks
期刊介绍: Annals of Clinical Microbiology and Antimicrobials considers good quality, novel and international research of more than regional relevance. Research must include epidemiological and/or clinical information about isolates, and the journal covers the clinical microbiology of bacteria, viruses and fungi, as well as antimicrobial treatment of infectious diseases. Annals of Clinical Microbiology and Antimicrobials is an open access, peer-reviewed journal focusing on information concerning clinical microbiology, infectious diseases and antimicrobials. The management of infectious disease is dependent on correct diagnosis and appropriate antimicrobial treatment, and with this in mind, the journal aims to improve the communication between laboratory and clinical science in the field of clinical microbiology and antimicrobial treatment. Furthermore, the journal has no restrictions on space or access; this ensures that the journal can reach the widest possible audience.
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