Fusobacterium nucleatum elicits subspecies-specific responses in human neutrophils.

IF 4.6 2区 医学 Q2 IMMUNOLOGY
Frontiers in Cellular and Infection Microbiology Pub Date : 2024-10-10 eCollection Date: 2024-01-01 DOI:10.3389/fcimb.2024.1449539
Maria Muchova, Sarah A Kuehne, Melissa M Grant, Peter P Smith, Malee Nagi, Iain L C Chapple, Josefine Hirschfeld
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引用次数: 0

Abstract

Fusobacterium nucleatum 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: animalis, fusiforme, nucleatum, polymorphum and vincentii. 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 F. nucleatum have not been studied to date. The aims of this study were to compare the immunogenicity of planktonic and biofilm-grown F. nucleatum and to investigate potential differences in human neutrophil responses when stimulated with individual F. nucleatum subspecies. Human neutrophils isolated from peripheral blood were stimulated with planktonic and biofilm-grown F. nucleatum 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 F. nucleatum led to significantly lower IL-1β release. We found significant differences among individual subspecies in terms of total, intracellular ROS and extracellular superoxide. Subspecies polymorphum 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 F. nucleatum in human neutrophils is highly subspecies-specific in vitro with regard to ROS release and cytokine production. Understanding subspecies-specific immunogenicity of F. nucleatum may facilitate the discovery of novel therapeutic targets in F. nucleatum-mediated diseases.

核分枝杆菌引起人类中性粒细胞亚种特异性反应
核酸镰刀菌是一种革兰氏阴性厌氧菌,在口腔生物膜中起着关键的桥接作用。它与牙周病和口腔外疾病有关,其中最主要的是大肠癌。它有五个亚种:animalis、fusiforme、nucleatum、polymorphum 和 vincentii。亚种与不断巡视组织的中性粒细胞相互作用,清除微生物入侵者。中性粒细胞的抗菌活动包括产生活性氧(ROS)、形成中性粒细胞胞外捕获物(NET)以及释放细胞因子和中性粒细胞酶。以前曾在中性粒细胞样细胞系中观察到免疫原性的亚种特异性差异,但没有在人类中性粒细胞中进行过研究。此外,迄今为止还没有研究过中性粒细胞对浮游生物和生物膜生长的 F. nucleatum 的反应。本研究的目的是比较浮游和生物膜生长的 F. nucleatum 的免疫原性,并研究人中性粒细胞在受到不同 F. nucleatum 亚种刺激时的潜在反应差异。用浮游和生物膜生长的 F. nucleatum 亚种刺激从外周血中分离出来的人类中性粒细胞。ROS的生成和NET的形成分别通过发光和荧光测定法进行量化。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的分泌通过酶联免疫吸附试验(ELISA)进行量化。中性粒细胞的反应显示,生物膜生长的细菌诱导的总 ROS 反应和细胞内 ROS 反应明显更高,总 ROS 释放时间也更短。生物膜生长的 F. nucleatum 导致 IL-1β 的释放明显降低。我们发现各个亚种在总ROS、细胞内ROS和细胞外超氧化物方面存在明显差异。多形亚种刺激的NET平均释放量最高。不同亚种释放的细胞因子数量差异显著,而溶酶体酶的释放量则没有差异。就 ROS 释放和细胞因子产生而言,F. nucleatum 在体外对人类中性粒细胞的免疫原性具有高度的亚种特异性。了解F. nucleatum的亚种特异性免疫原性有助于发现F. nucleatum介导的疾病的新型治疗靶点。
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来源期刊
CiteScore
7.90
自引率
7.00%
发文量
1817
审稿时长
14 weeks
期刊介绍: Frontiers in Cellular and Infection Microbiology is a leading specialty journal, publishing rigorously peer-reviewed research across all pathogenic microorganisms and their interaction with their hosts. Chief Editor Yousef Abu Kwaik, University of Louisville is supported by an outstanding Editorial Board of international experts. This multidisciplinary open-access journal is at the forefront of disseminating and communicating scientific knowledge and impactful discoveries to researchers, academics, clinicians and the public worldwide. Frontiers in Cellular and Infection Microbiology includes research on bacteria, fungi, parasites, viruses, endosymbionts, prions and all microbial pathogens as well as the microbiota and its effect on health and disease in various hosts. The research approaches include molecular microbiology, cellular microbiology, gene regulation, proteomics, signal transduction, pathogenic evolution, genomics, structural biology, and virulence factors as well as model hosts. Areas of research to counteract infectious agents by the host include the host innate and adaptive immune responses as well as metabolic restrictions to various pathogenic microorganisms, vaccine design and development against various pathogenic microorganisms, and the mechanisms of antibiotic resistance and its countermeasures.
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