Periodontal Research Group, Birmingham School of Dentistry, Institute of Clinical Sciences, The University of Birmingham, Birmingham, United Kingdom.
School of Science and Technology, Nottingham Trent University, Nottingham, United Kingdom.
Front Cell Infect Microbiol. 2024 Oct 10;14:1449539. doi: 10.3389/fcimb.2024.1449539. eCollection 2024.
as a Gram-negative anaerobe plays a key bridging role in oral biofilms. It is involved in periodontal and extraoral diseases, the most prominent being colorectal cancer. Five subspecies are recognised: and . Subspecies interact with neutrophils constantly patrolling tissues to remove microbial intruders. Neutrophil antimicrobial activities include generation of reactive oxygen species (ROS), formation of neutrophil extracellular traps (NETs) and release of cytokines and neutrophil enzymes. Subspecies-specific differences in immunogenicity have previously been observed in a neutrophil-like cell line but were not investigated in human neutrophils. Additionally, neutrophil responses to planktonic and biofilm-grown have not been studied to date. The aims of this study were to compare the immunogenicity of planktonic and biofilm-grown and to investigate potential differences in human neutrophil responses when stimulated with individual subspecies. Human neutrophils isolated from peripheral blood were stimulated with planktonic and biofilm-grown subspecies. Generation of ROS and NET formation were quantified by luminescence and fluorescence assays, respectively. Secretion of cytokines (IL-1β, TNF-α, IL-6, IL-8), neutrophil elastase and matrix metalloproteinase-9 was quantified by enzyme-linked immunosorbent assay (ELISA). Neutrophil responses showed biofilm-grown bacteria induced a significantly higher total and intracellular ROS response, as well as shorter time to total ROS release. Biofilm-grown led to significantly lower IL-1β release. We found significant differences among individual subspecies in terms of total, intracellular ROS and extracellular superoxide. Subspecies stimulated the highest mean amount of NET release. Amounts of cytokines released differed significantly among subspecies, while no differences were found in lysosomal enzyme release. Immunogenicity of in human neutrophils is highly subspecies-specific with regard to ROS release and cytokine production. Understanding subspecies-specific immunogenicity of may facilitate the discovery of novel therapeutic targets in -mediated diseases.
作为一种革兰氏阴性厌氧菌，它在口腔生物膜中起着关键的桥梁作用。它与牙周病和口腔外疾病有关，其中最突出的是结直肠癌。目前已识别出 5 个亚种： 和 。亚种与不断巡逻组织以清除微生物入侵者的中性粒细胞相互作用。中性粒细胞的抗菌活性包括产生活性氧物质（ROS）、形成中性粒细胞细胞外陷阱（NETs）以及释放细胞因子和中性粒细胞酶。以前在中性粒细胞样细胞系中观察到亚种特异性的免疫原性差异，但在人类中性粒细胞中尚未进行研究。此外，迄今为止尚未研究浮游和生物膜生长的 对中性粒细胞的反应。本研究的目的是比较浮游和生物膜生长的 的免疫原性，并研究当用单个亚种刺激时人类中性粒细胞反应的潜在差异。从外周血中分离出人类中性粒细胞，用浮游和生物膜生长的 亚种刺激。通过发光和荧光测定分别定量 ROS 的产生和 NET 的形成。通过酶联免疫吸附试验（ELISA）定量细胞因子（IL-1β、TNF-α、IL-6、IL-8）、中性粒细胞弹性蛋白酶和基质金属蛋白酶-9 的分泌。中性粒细胞反应表明，生物膜生长的细菌诱导的总和细胞内 ROS 反应更高，总 ROS 释放的时间更短。生物膜生长的 导致 IL-1β 的释放显著降低。我们发现单个亚种之间在总 ROS、细胞内 ROS 和细胞外超氧化物方面存在显著差异。亚种 刺激的 NET 释放量最高。释放的细胞因子量在亚种之间存在显著差异，而溶酶体酶释放没有差异。 在人类中性粒细胞中的免疫原性高度依赖于亚种，涉及 ROS 释放和细胞因子产生。了解 亚种特异性免疫原性可能有助于发现 介导疾病的新治疗靶点。
