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Vaginolysin Drives Epithelial Ultrastructural Responses to Gardnerella vaginalis

机译：阴道溶素驱动阴道加德纳菌的上皮超微结构反应

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Gardnerella vaginalis, the bacterial species most frequently isolated from women with bacterial vaginosis (BV), produces a cholesterol-dependent cytolysin (CDC), vaginolysin (VLY). At sublytic concentrations, CDCs may initiate complex signaling cascades crucial to target cell survival. Using live-cell imaging, we observed the rapid formation of large membrane blebs in human vaginal and cervical epithelial cells (VK2 and HeLa cells) exposed to recombinant VLY toxin and to cell-free supernatants from growing liquid cultures of G. vaginalis. Binding of VLY to its human-specific receptor (hCD59) is required for bleb formation, as antibody inhibition of either toxin or hCD59 abrogates this response, and transfection of nonhuman cells (CHO-K1) with hCD59 renders them susceptible to toxin-induced membrane blebbing. Disruption of the pore formation process (by exposure to pore-deficient toxoids or pretreatment of cells with methyl-β-cyclodextrin) or osmotic protection of target cells inhibits VLY-induced membrane blebbing. These results indicate that the formation of functional pores drives the observed ultrastructural rearrangements. Rapid bleb formation may represent a conserved response of epithelial cells to sublytic quantities of pore-forming toxins, and VLY-induced epithelial cell membrane blebbing in the vaginal mucosa may play a role in the pathogenesis of BV.
机译：阴道加德纳菌是最常从患有细菌性阴道病（BV）的女性中分离出的细菌，可产生胆固醇依赖性细胞溶素（CDC），阴道溶素（VLY）。在分解浓度下，CDC可能会启动对靶细胞存活至关重要的复杂信号传导级联。使用活细胞成像，我们观察到暴露于重组VLY毒素和生长自阴道加德纳斯菌的无细胞上清液的人阴道和宫颈上皮细胞（VK2和HeLa细胞）中大的膜泡迅速形成。气泡形成需要VLY与其人类特异性受体（hCD59）结合，因为对毒素或hCD59的抗体抑制作用会消除这种反应，并且用hCD59转染非人类细胞（CHO-K1）使其易于受到毒素诱导的膜的影响起泡。破坏孔形成过程（通过暴露于缺乏孔的类毒素或用甲基-β-环糊精预处理细胞）或对靶细胞进行渗透保护可抑制VLY诱导的膜起泡。这些结果表明功能孔的形成驱动观察到的超微结构重排。气泡的快速形成可能代表上皮细胞对亚溶解量的成孔毒素的保守反应，阴道黏膜中VLY诱导的上皮细胞膜起泡可能在BV的发病机理中起作用。

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