Mouzykantov A A, Rozhina E V, Fakhrullin R F, Gomzikova M O, Zolotykh M A, Chernova O A, Chernov V M
Kazan Institute of Biochemistry and Biophysics, FRC Kazan Scientific Center of RAS, Kazan, 420111 Russia.
Kazan Federal University, Kazan, 420008 Russia.
Acta Naturae. 2021 Oct-Dec;13(4):82-88. doi: 10.32607/actanaturae.11506.
The extracellular vesicles (EVs) produced by bacteria transport a wide range of compounds, including proteins, DNA and RNA, mediate intercellular interactions, and may be important participants in the mechanisms underlying the persistence of infectious agents. This study focuses on testing the hypothesis that the EVs of mycoplasmas, the smallest prokaryotes capable of independent reproduction, combined in the class referred to as Mollicutes, can penetrate into eukaryotic cells and modulate their immunoreactivity. To verify this hypothesis, for the first time, studies of in vitro interaction between human skin fibroblasts and vesicles isolated from Acholeplasma laidlawii (the ubiquitous mycoplasma that infects higher eukaryotes and is the main contaminant of cell cultures and vaccines) were conducted using confocal laser scanning microscopy and proteome profiling, employing a combination of 2D-DIGE and MALDI-TOF/TOF, the Mascot mass-spectrum analysis software and the DAVID functional annotation tool. These studies have revealed for the first time that the extracellular vesicles of A. laidlawii can penetrate into eukaryotic cells in vitro and modulate the expression of cellular proteins. The molecular mechanisms behind the interaction of mycoplasma vesicles with eukaryotic cells and the contribution of the respective nanostructures to the molecular machinery of cellular permissiveness still remain to be elucidated. The study of these aspects is relevant both for fundamental research into the "logic of life" of the simplest prokaryotes, and the practical development of efficient control over hypermutable bacteria infecting humans, animals and plants, as well as contaminating cell cultures and vaccines.
细菌产生的细胞外囊泡（EVs）可运输多种化合物，包括蛋白质、DNA和RNA，介导细胞间相互作用，并且可能是感染因子持续存在机制的重要参与者。本研究着重验证以下假设：支原体（能够独立繁殖的最小原核生物，归为柔膜菌纲）的细胞外囊泡能够穿透真核细胞并调节其免疫反应性。为验证这一假设，首次使用共聚焦激光扫描显微镜和蛋白质组分析方法，结合二维差异凝胶电泳（2D-DIGE）和基质辅助激光解吸电离飞行时间质谱/串联质谱（MALDI-TOF/TOF）、Mascot质谱分析软件和DAVID功能注释工具，对人皮肤成纤维细胞与从莱氏无胆甾原体（一种普遍存在的支原体，可感染高等真核生物，是细胞培养物和疫苗的主要污染物）分离出的囊泡之间的体外相互作用进行了研究。这些研究首次揭示，莱氏无胆甾原体的细胞外囊泡能够在体外穿透真核细胞并调节细胞蛋白质的表达。支原体囊泡与真核细胞相互作用背后的分子机制以及相应纳米结构对细胞易感性分子机制的贡献仍有待阐明。对这些方面的研究，对于最简单原核生物“生命逻辑”的基础研究，以及对感染人类、动物和植物以及污染细胞培养物和疫苗的超突变细菌进行有效控制的实际开发都具有重要意义。
