Multiparametric sensing of outer membrane vesicle-derived supported lipid bilayers demonstrates the specificity of bacteriophage interactions
Multiparametric sensing of outer membrane vesicle-derived supported lipid bilayers demonstrates the specificity of bacteriophage interactions
复制标题
外膜囊泡衍生的支持的脂质双层的多参数传感证明了噬菌体相互作用的特异性
DOI：
10.1101/2022.12.13.520201
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发表时间：
2022
期刊：
影响因子：
--
通讯作者：
Bali K
中科院分区：
文献类型：
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作者：
Bali K
近年来，随着传统抗生素的有效性下降，使用噬菌体（一种专门感染细菌的病毒）作为抗生素已成为人们非常感兴趣的领域。快速定量地检测噬菌体与特定细菌的相互作用是鉴定新型抗菌剂感兴趣的噬菌体的关键。来源于革兰氏阴性菌的外膜囊泡（omv）可用于制造支持脂质双分子层（slb），因此可用于含有细菌外膜天然成分的体外膜模型。在这项研究中，我们采用大肠杆菌衍生的slb，并使用荧光成像和机械传感技术来显示它们与T4噬菌体的相互作用。我们还将这些双分子层与导电聚合物PEDOT:PSS功能化的微电极阵列（MEAs）集成，并表明可以使用电阻抗谱来监测噬菌体与slb的孔形成相互作用。为了突出我们检测特定噬菌体相互作用的能力，我们还使用从啮齿类微杆菌（mcitrobacter rodentium）中提取的omv生成了slb，该细菌对T4噬菌体感染具有抗性，并确定它们与噬菌体缺乏相互作用。这里提出的工作表明，如何相互作用发生在噬菌体和这些复杂的SLB系统之间，可以使用一系列的实验技术进行监测。我们相信，这种方法可以用来识别噬菌体，可以对抗感兴趣的细菌菌株，也可以更广泛地用于监测与细菌外膜相互作用的任何孔形成结构（如防御素），从而有助于开发下一代抗菌剂。
The use of bacteriophages, viruses that specifically infect bacteria, as antibiotics has become an area of great interest in recent years as the effectiveness of conventional antibiotics recedes. The detection of phage interactions with specific bacteria in a rapid and quantitative way is key for identifying phages of interest for novel antimicrobials. Outer membrane vesicles (OMVs) derived from Gram-negative bacteria can be used to make supported lipid bilayers (SLBs) and thereforein vitromembrane models that contain naturally occurring components of the bacterial outer membrane. In this study, we employedEscherichia coliOMV derived SLBs and use both fluorescent imaging and mechanical sensing techniques to show their interactions with T4 phage. We also integrate these bilayers with microelectrode arrays (MEAs) functionalized with the conducting polymer PEDOT:PSS and show that the pore forming interactions of the phages with the SLBs can be monitored using electrical impedance spectroscopy. To highlight our ability to detect specific phage interactions, we also generate SLBs using OMVs derived fromCitrobacter rodentium, which is resistant to T4 phage infection, and identify their lack of interaction with the phage. The work presented here shows how interactions occurring between the phages and these complex SLB systems can be monitored using a range of experimental techniques. We believe this approach can be used to identify phages that work against bacterial strains of interest, as well as more generally to monitor any pore forming structure (such as defensins) interacting with bacterial outer membranes, and thus aid in the development of next generation antimicrobials.