Abstract：
Bacterial extracellular vesicles (BEVs) are nano- or micrometer-sized membrane-bound lipid vesicles released from both Gram-negative and Gram-positive bacteria. Cellular transport, communication, pathogenesis, and host-pathogen interactions are some of the major biological processes impacted by BEVs. Among these, host-pathogen interactions and bacterial pathogenesis are emerging as highly important targetable avenues underlined by the issues of antimicrobial resistance, thus demanding novel targets and approaches to treat bacterial infections. In this aspect, the study of the interaction of BEVs with bacteria and/or host cells becomes imperative and brings the membrane fusion process to the forefront. Furthermore, membrane fusion also underscores the performance of BEVs as nano-therapeutic delivery platforms. Here, we report methods to study fusion kinetics between mycobacteria-derived extracellular vesicles, which we refer to as MEVs, and intact mycobacteria or MEVs themselves. We also discuss the isolation of MEVs and their characterization. We outline critical factors that affect fusion kinetics by MEVs. The same principle can be extended for studying fusion between BEVs and mammalian host cells important for understanding how BEVs influence host-pathogen crosstalk.
摘要：
细菌细胞外囊泡(BEV)是从革兰氏阴性和革兰氏阳性细菌释放的纳米或微米大小的膜结合的脂质囊泡。蜂窝运输，通信,发病机制，和宿主-病原体相互作用是BEV影响的一些主要生物过程。其中,宿主-病原体相互作用和细菌发病机制正在成为抗生素耐药性问题强调的非常重要的可靶向途径，因此需要新的目标和方法来治疗细菌感染。在这方面,研究BEV与细菌和/或宿主细胞的相互作用变得势在必行，并将膜融合过程带到最前沿。此外，膜融合也强调了BEV作为纳米治疗递送平台的性能。这里,我们报告了研究分枝杆菌来源的细胞外囊泡之间融合动力学的方法，我们称之为MEV，和完整的分枝杆菌或MEV本身。我们还讨论了MEV的分离及其表征。我们概述了影响MEV融合动力学的关键因素。相同的原理可以扩展用于研究BEV与哺乳动物宿主细胞之间的融合，这对于理解BEV如何影响宿主-病原体串扰很重要。
