Architecture and matrix assembly determinants of Bordetella pertussis biofilms on primary human airway epithelium.
Architecture and matrix assembly determinants of Bordetella pertussis biofilms on primary human airway epithelium.
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DOI：
10.1371/journal.ppat.1011193
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发表时间：
2023-03
期刊：
影响因子：
6.7
通讯作者：
中科院分区：
文献类型：
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传统上，由专性人类病原体百日咳博德泰拉（Bp）引起的百日咳或百日咳被描述为具有严重症状的急性疾病。然而，许多感染百日咳的人要么没有症状，要么表现出非常轻微的症状，但却可能成为细菌传播的携带者和来源。生物膜是细菌在人类感染和疾病中的重要生存机制。然而，驱动人类生物膜形成的细菌决定因素尚不明确。在目前的研究中，我们发现Bp感染分化良好的原代人支气管上皮细胞导致细菌聚集、簇和高度结构化的生物膜的形成，这些生物膜与纤毛共定位。这些发现与百日咳患者组织病理分析的观察结果相似。多糖Bps、细胞外DNA和细菌细胞的不同排列（单、双和三方）被可视化，表明细菌-基质相互作用的复杂异质性。突变生物膜的分析揭示了三种关键Bp毒力因子（Bp、丝状血凝素和腺苷酸环化酶毒素）在基质生产、细胞簇形成和生物膜成熟度方面的积极作用。黏附实验证实Bps是一种新的Bp黏附素，适用于原代人气道细胞。综上所述，我们的研究结果证明了生物膜细胞外基质和生物膜在模拟人类呼吸道条件下发育过程的多因子性质，并强调了类似自然宿主环境的模型系统对研究发病机制和潜在治疗策略的重要性。尽管广泛接种疫苗，但由革兰氏阴性专性人类病原体百日咳博德泰拉（Bp）引起的百日咳在许多国家正在死灰复燃。Bp感染人类宿主、引起疾病并在鼻咽部持续存在的机制尚不明确。我们在此表明，Bp在分化良好和有纤毛的原代人支气管上皮细胞（HBE）上与纤毛共定位形成聚集体、簇和高度结构化的生物膜。HBE细胞上的生物膜细胞外基质（ECM）是异质性的，由多糖Bps、细胞外DNA和细菌细胞组成，它们以不同的复合物（单、双、三方）排列。利用突变菌株，我们发现了三种Bp毒力因子，Bps，丝状血凝素和腺苷酸环化酶毒素在基质产生，细胞簇形成和生物膜成熟中的积极作用。我们的研究为研究细菌生物膜提供了一个与人类相关的模型，揭示了ECM的本质以及在人气道细胞上形成生物膜所需的细菌决定因素。
Traditionally, whooping cough or pertussis caused by the obligate human pathogen Bordetella pertussis (Bp) is described as an acute disease with severe symptoms. However, many individuals who contract pertussis are either asymptomatic or show very mild symptoms and yet can serve as carriers and sources of bacterial transmission. Biofilms are an important survival mechanism for bacteria in human infections and disease. However, bacterial determinants that drive biofilm formation in humans are ill-defined. In the current study, we show that Bp infection of well-differentiated primary human bronchial epithelial cells leads to formation of bacterial aggregates, clusters, and highly structured biofilms which are colocalized with cilia. These findings mimic observations from pathological analyses of tissues from pertussis patients. Distinct arrangements (mono-, bi-, and tri-partite) of the polysaccharide Bps, extracellular DNA, and bacterial cells were visualized, suggesting complex heterogeneity in bacteria-matrix interactions. Analyses of mutant biofilms revealed positive roles in matrix production, cell cluster formation, and biofilm maturity for three critical Bp virulence factors: Bps, filamentous hemagglutinin, and adenylate cyclase toxin. Adherence assays identified Bps as a new Bp adhesin for primary human airway cells. Taken together, our results demonstrate the multi-factorial nature of the biofilm extracellular matrix and biofilm development process under conditions mimicking the human respiratory tract and highlight the importance of model systems resembling the natural host environment to investigate pathogenesis and potential therapeutic strategies. Despite widespread vaccination, pertussis or whooping cough caused by the gram-negative obligate human pathogen Bordetella pertussis (Bp) is resurging in many countries. The mechanisms by which Bp infects human hosts, causes disease, and persists in the nasopharynx are ill-defined. We show herein that Bp forms aggregates, clusters, and highly structured biofilms colocalized with cilia on well-differentiated and ciliated primary human bronchial epithelial (HBE) cells. The biofilm extracellular matrix (ECM) on HBE cells was heterogenous and consisted of the polysaccharide Bps, extracellular DNA, and bacterial cells arranged in different complexes (mono-, bi-, and tri-partite). Utilizing mutant strains, we discovered positive roles of three Bp virulence factors, Bps, filamentous hemagglutinin, and adenylate cyclase toxin in matrix production, cell cluster formation and biofilm maturation. Our study provides a human-relevant model for studying bacterial biofilms, reveals the nature of ECM and bacterial determinants required for biofilm formation on primary human airway cells.
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影响因子：
7.7
作者：
Gill CJ;Gunning CE;MacLeod WB;Mwananyanda L;Thea DM;Pieciak RC;Kwenda G;Mupila Z;Rohani P
通讯作者：
Rohani P
影响因子：
5.2
作者：
Belcher T;Dubois V;Rivera-Millot A;Locht C;Jacob-Dubuisson F
通讯作者：
Jacob-Dubuisson F
DOI：
10.1126/science.1222981
发表时间：
2012-07-13
期刊：
Science (New York, N.Y.)
影响因子：
--
作者：
Berk V;Fong JC;Dempsey GT;Develioglu ON;Zhuang X;Liphardt J;Yildiz FH;Chu S
通讯作者：
Chu S
影响因子：
3.1
作者：
COOKSON, BT;CHO, HL;GOLDMAN, WE
通讯作者：
GOLDMAN, WE
影响因子：
3.1
作者：
Eby, Joshua C.;Gray, Mary C.;Hewlett, Erik L.
通讯作者：
Hewlett, Erik L.