Rosenfeld Lea, Neumann Nico, Bao Xin, Adam Aysegül, Schaefer Arne S
Department of Periodontology, Oral Surgery and Oral Medicine, Universitätsmedizin Berlin, Berlin, Germany.
Infect Immun. 2025 Apr 8;93(4):e0000525. doi: 10.1128/iai.00005-25. Epub 2025 Mar 21.
The protozoan commonly colonizes anaerobic periodontal pockets, induces a severe innate immune response, invades gingival mucosa, and kills epithelial cells. infection is associated with the common oral inflammatory disease periodontitis. DNA variants in vesicle-associated membrane proteins (VAMP) -3 and -8 genes are linked to increased periodontitis risk. These genes mediate host-pathogen interactions, including mucin exocytosis to form protective barriers and matrix metalloproteinase (MMP) secretion in intestinal amoebiasis caused by . This study aimed to investigate the roles of VAMP3/8 in gingival defense and infection mechanisms. Clustered regularly interspaced short palindromic repeats (CRISPR)-Cas9 gene editing was used to create VAMP3/8-deficient gingival epithelial cells and fibroblasts. Functional analyses included immunofluorescence, enzyme-linked immunosorbent assay (ELISA), cytotoxicity, and collagenase assays. VAMP8 co-localized with mucins in gingival epithelial cells (gECs), and VAMP3 with MMPs in gingival fibroblasts. In gECs infection increased mucin (MUC1: 3.6×, MUC21: 14.4×) and interleukin secretion (IL-8, IL-1B: >6×, = 0.019). deficiency in gECs caused higher cell death (35% vs 4% in controls) with reduced exocytosis of mucins and interleukins. Likewise, -induced VAMP8 translocation into lipid rafts was lost in VAMP8 knockout cells, validating the participation of VAMP8 in exocytosis. In wild-type but not VAMP3-deficient gingival fibroblasts, strongly activated collagenases. effects were more pathogenic than those of the oral anaerobic bacterium exploits VAMP8/3-driven exocytosis pathways, driving inflammation and tissue destruction, underscoring its role as a significant periodontal pathogen.
这种原生动物通常定殖于厌氧性牙周袋，引发严重的先天性免疫反应，侵入牙龈黏膜并杀死上皮细胞。感染与常见的口腔炎性疾病牙周炎相关。囊泡相关膜蛋白（VAMP）-3和-8基因中的DNA变异与牙周炎风险增加有关。这些基因介导宿主与病原体的相互作用，包括在由[具体病原体名称未给出]引起的肠道阿米巴病中通过粘蛋白胞吐作用形成保护屏障以及分泌基质金属蛋白酶（MMP）。本研究旨在探究VAMP3/8在牙龈防御和[具体病原体名称未给出]感染机制中的作用。利用成簇规律间隔短回文重复序列（CRISPR）-Cas9基因编辑技术创建VAMP3/8缺陷型牙龈上皮细胞和成纤维细胞。功能分析包括免疫荧光、酶联免疫吸附测定（ELISA）、细胞毒性和胶原酶测定。VAMP8在牙龈上皮细胞（gECs）中与粘蛋白共定位，VAMP3在牙龈成纤维细胞中与MMP共定位。在gECs中，[具体病原体名称未给出]感染增加了粘蛋白（MUC1：3.6倍，MUC21：14.4倍）和白细胞介素分泌（IL-8、IL-1β：>6倍，P = 0.019）。gECs中的[具体病原体名称未给出]缺陷导致更高的细胞死亡（35%对比对照组的4%），同时粘蛋白和白细胞介素的胞吐作用减少。同样，[具体病原体名称未给出]诱导的VAMP8易位到脂筏在VAMP8基因敲除细胞中消失，证实了VAMP8参与胞吐作用。在野生型而非VAMP3缺陷型牙龈成纤维细胞中，[具体病原体名称未给出]强烈激活胶原酶。[具体病原体名称未给出]的作用比口腔厌氧菌更具致病性，[具体病原体名称未给出]利用VAMP8/3驱动的胞吐途径，引发炎症和组织破坏，突出了其作为重要牙周病原体的作用。
