Periodontitis is a chronic inflammatory disease caused by periodontal pathogens in subgingival plaque and is associated with systemic inflammatory diseases. Extracellular vesicles (EVs) released from host cells and pathogens carry a variety of biological molecules and are of interest for their role in disease progression and as diagnostic markers. In the present study, we analysed the proteome and inflammatory response of EVs derived from macrophages infected with Tannerella forsythia, a periodontal pathogen. The EVs isolated from the cell conditioned medium of T. forsythia-infected macrophages were divided into two distinct vesicles, macrophage-derived EVs and T. forsythia-derived OMVs, by size exclusion chromatography combined with density gradient ultracentrifugation. Proteome analysis showed that in T. forsythia infection, macrophage-derived EVs were enriched with pro-inflammatory cytokines and inflammatory mediators associated with periodontitis progression. T. forsythia-derived OMVs harboured several known virulence factors, including BspA, sialidase, GroEL and various bacterial lipoproteins. T. forsythia-derived OMVs induced pro-inflammatory responses via TLR2 activation. In addition, we demonstrated that T. forsythia actively released OMVs when T. forsythia encountered macrophage-derived soluble molecules. Taken together, our results provide insight into the characterisation of EVs derived from cells infected with a periodontal pathogen.

中文翻译：

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感染牙周病原菌连翘的巨噬细胞衍生的细胞外囊泡的蛋白质组和免疫反应

牙周炎是由龈下菌斑内的牙周病原菌引起的慢性炎症性疾病，与全身炎症性疾病有关。从宿主细胞和病原体释放的细胞外囊泡（EV）携带多种生物分子，因其在疾病进展中的作用和作为诊断标记物而引起人们的兴趣。在本研究中，我们分析了感染牙周病原体福赛坦纳菌的巨噬细胞衍生的 EV 的蛋白质组和炎症反应。通过尺寸排阻色谱结合密度梯度超速离心，从连翘感染的巨噬细胞的细胞条件培养基中分离的EV被分成两个不同的囊泡，巨噬细胞来源的EV和连翘来源的OMV。蛋白质组分析表明，在连翘感染中，巨噬细胞衍生的 EV 富含与牙周炎进展相关的促炎细胞因子和炎症介质。连翘衍生的 OMV 含有多种已知的毒力因子，包括 BspA、唾液酸酶、GroEL 和各种细菌脂蛋白。连翘衍生的 OMV 通过 TLR2 激活诱导促炎反应。此外，我们还证明，当连翘遇到巨噬细胞衍生的可溶性分子时，连翘会主动释放 OMV 。总而言之，我们的结果提供了对牙周病原体感染细胞衍生的 EV 特征的深入了解。