Filifactor alocis, a gram‐positive, obligate anaerobic rod, is an emerging periodontal pathogen that is frequently isolated from patients with periodontitis, peri‐implantitis, and apical periodontitis. Recent studies have shown that extracellular vesicles (EVs) from gram‐negative periodontal pathogens, so‐called outer membrane vesicles (OMVs), harbor various effector molecules responsible for inducing host inflammatory responses. However, there are no reports of EVs from F. alocis. In this study, we purified and characterized the protein profiles of EVs from F. alocis and investigated their immunostimulatory activity on human monocytic THP‐1 and human oral keratinocyte HOK‐16B cell lines. Highly pure EVs were obtained from F. alocis using density gradient ultracentrifugation. Nanoparticle tracking analysis and transmission electron microscopy showed that F. alocis EVs were between 50 and 270 nm in diameter. Proteome analysis identified 28 proteins, including lipoproteins, autolysins, F. alocis complement inhibitor (FACIN), transporter‐related proteins, metabolism‐related proteins, and ribosomal proteins. Human cytokine array analysis showed that F. alocis EVs remarkably induced the expression of CCL1, CCL2, MIP‐1, CCL5, CXCL1, CXCL10, ICAM‐1, IL‐1β, IL‐1ra, IL‐6, IL‐8, MIF, SerpinE, and TNF‐α in THP‐1 cells and CXCL1, G‐CSF, GM‐CSF, IL‐6, and IL‐8 in HOK‐16B cells. The immunostimulatory activity of F. alocis EVs was similar to that of the whole bacterial cells. Our findings provide new insight into the role of EVs from gram‐positive oral bacteria in periodontal diseases.

中文翻译：

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Filifactor alocis胞外囊泡的表征和免疫刺激活性。

纤毛变形虫，革兰氏阳性，专性厌氧棒，是一种新兴的牙周病原体，通常从患有牙周炎，种植体周围炎和根尖周炎的患者中分离出来。最近的研究表明，来自革兰氏阴性牙周病原体的细胞外囊泡（EVs），即所谓的外膜囊泡（OMVs），具有各种可诱导宿主炎症反应的效应分子。但是，目前还没有关于洋芋F的报道。在这项研究中，我们纯化和表征了来自海芋的EV的蛋白质谱，并研究了它们对人单核细胞THP-1和人口腔角质形成细胞HOK-16B细胞系的免疫刺激活性。高纯电动汽车获自海芋使用密度梯度超速离心。纳米粒子跟踪分析和透射电子显微镜显示，海芋F的直径在50至270 nm之间。蛋白质组学分析确定了28种蛋白质，包括脂蛋白，自溶素，海芋F.INC补体抑制剂（FACIN），转运蛋白相关蛋白，代谢相关蛋白和核糖体蛋白。人细胞因子阵列分析显示，海芋EV明显诱导CCL1，CCL2，MIP-1，CCL5，CXCL1，CXCL10，ICAM-1，IL-1β，IL-1ra，IL-6，IL-8，MIF的表达，THP-1细胞中的SerpinE和TNF-α以及HOK-16B细胞中的CXCL1，G-CSF，GM-CSF，IL-6和IL-8。海芋的免疫刺激活性电动汽车类似于整个细菌细胞。我们的发现为革兰氏阳性口腔细菌的电动车在牙周疾病中的作用提供了新的见解。