重症哮喘微生物免疫相互作用的物种水平、宏基因组和蛋白质组学分析
2024/08/28
摘要
背景：重症哮喘的气道微生物组尚未通过宏基因组测序在物种水平上进行表征，且“低2型”中性粒细胞哮喘中特定物种与粘膜免疫反应之间的关系也尚未明确。本研究采用炎症介质进行综合物种水平宏基因组数据分析，以表征显性潜在致病原和宿主免疫反应的流行情况。
方法：在Wessex（n=66）和Oxford（n=30）两个横断面成人重度哮喘队列中，采用Nanopore和qPCR的长读宏基因组测序对痰液和鼻腔灌洗液样本进行分析。本研究将物种水平的数据与临床参数以及通过免疫测定和O-link测量的39种选定的气道蛋白进行了整合。
结果：健康和轻度哮喘患者的痰液微生物组显示出高微生物多样性。相比之下，23%（19/81）的严重哮喘患者的微生物组由一种呼吸道病原体主导，即流感嗜血杆菌（n=10）、卡他莫拉菌（n=4）、肺炎链球菌（=4）和铜绿假单胞菌（n=1）。中性粒细胞哮喘与流感嗜血杆菌、卡他莫拉菌、肺炎链球菌、惠普尔养障体相关，表现为1型细胞因子和蛋白酶升高；嗜酸性粒细胞性哮喘患者中，卡他莫拉菌含量较高，但流感嗜血杆菌和肺炎链球菌含量较低。流感嗜血杆菌载量与嗜酸性粒细胞阳离子蛋白、弹性蛋白酶和IL-10相关。黏液罗氏菌与IL-6呈正相关，与FGF呈负相关。贝叶斯网络分析还揭示了流感嗜血杆菌和卡他莫拉菌与1型气道炎症密切且显著相关。上、下呼吸道的微生物组和细胞因子环境不同。
结论：物种水平的综合分析揭示了潜在致病菌与严重哮喘气道炎症之间的主要且不同的相关性。
Jabeen MF, Sanderson ND, Tinè M, Donachie G, Barber C, Azim A, Lau LCK, Brown T, Pavord ID, Chauhan A, Klenerman P, Street TL, Marchi E, Howarth PH, Hinks TSC.
Abstract
BACKGROUND:The airway microbiome in severe asthma has not been characterised at species-level by metagenomic sequencing, nor have the relationships between specific species and mucosal immune responses in 'type-2 low', neutrophilic asthma been defined. We performed an integrated species-level metagenomic data with inflammatory mediators to characterise prevalence of dominant potentially pathogenic organisms and host immune responses.
METHODS:Sputum and nasal lavage samples were analysed using long-read metagenomic sequencing with Nanopore and qPCR in two cross-sectional adult severe asthma cohorts, Wessex (n = 66) and Oxford (n = 30). We integrated species-level data with clinical parameters and 39 selected airway proteins measured by immunoassay and O-link.
RESULTS:The sputum microbiome in health and mild asthma displayed comparable microbial diversity. By contrast, 23% (19/81) of severe asthma microbiomes were dominated by a single respiratory pathogen, namely H. influenzae (n = 10), M. catarrhalis (n = 4), S. pneumoniae (n = 4) and P. aeruginosa (n = 1). Neutrophilic asthma was associated with H. influenzae, M. catarrhalis, S. pneumoniae and T. whipplei with elevated type-1 cytokines and proteases; eosinophilic asthma with higher M. catarrhalis, but lower H. influenzae, and S. pneumoniae abundance. H. influenzae load correlated with Eosinophil Cationic Protein, elastase and IL-10. R. mucilaginosa associated positively with IL-6 and negatively with FGF. Bayesian network analysis also revealed close and distinct relationships of H. influenzae and M. catarrhalis with type-1 airway inflammation. The microbiomes and cytokine milieu were distinct between upper and lower airways.
CONCLUSION:This species-level integrated analysis reveals central, but distinct associations between potentially pathogenic bacteria and airways inflammation in severe asthma.
背景：重症哮喘的气道微生物组尚未通过宏基因组测序在物种水平上进行表征，且“低2型”中性粒细胞哮喘中特定物种与粘膜免疫反应之间的关系也尚未明确。本研究采用炎症介质进行综合物种水平宏基因组数据分析，以表征显性潜在致病原和宿主免疫反应的流行情况。
方法：在Wessex（n=66）和Oxford（n=30）两个横断面成人重度哮喘队列中，采用Nanopore和qPCR的长读宏基因组测序对痰液和鼻腔灌洗液样本进行分析。本研究将物种水平的数据与临床参数以及通过免疫测定和O-link测量的39种选定的气道蛋白进行了整合。
结果：健康和轻度哮喘患者的痰液微生物组显示出高微生物多样性。相比之下，23%（19/81）的严重哮喘患者的微生物组由一种呼吸道病原体主导，即流感嗜血杆菌（n=10）、卡他莫拉菌（n=4）、肺炎链球菌（=4）和铜绿假单胞菌（n=1）。中性粒细胞哮喘与流感嗜血杆菌、卡他莫拉菌、肺炎链球菌、惠普尔养障体相关，表现为1型细胞因子和蛋白酶升高；嗜酸性粒细胞性哮喘患者中，卡他莫拉菌含量较高，但流感嗜血杆菌和肺炎链球菌含量较低。流感嗜血杆菌载量与嗜酸性粒细胞阳离子蛋白、弹性蛋白酶和IL-10相关。黏液罗氏菌与IL-6呈正相关，与FGF呈负相关。贝叶斯网络分析还揭示了流感嗜血杆菌和卡他莫拉菌与1型气道炎症密切且显著相关。上、下呼吸道的微生物组和细胞因子环境不同。
结论：物种水平的综合分析揭示了潜在致病菌与严重哮喘气道炎症之间的主要且不同的相关性。
(中日友好医院呼吸与危重症医学科 张婧媛 摘译 林江涛 审校)
(Allergy. 2024 Aug 11. doi: 10.1111/all.16269.)
Species-level, metagenomic and proteomic analysis of microbe-immune interactions in severe asthma.(Allergy. 2024 Aug 11. doi: 10.1111/all.16269.)
Jabeen MF, Sanderson ND, Tinè M, Donachie G, Barber C, Azim A, Lau LCK, Brown T, Pavord ID, Chauhan A, Klenerman P, Street TL, Marchi E, Howarth PH, Hinks TSC.
Abstract
BACKGROUND:The airway microbiome in severe asthma has not been characterised at species-level by metagenomic sequencing, nor have the relationships between specific species and mucosal immune responses in 'type-2 low', neutrophilic asthma been defined. We performed an integrated species-level metagenomic data with inflammatory mediators to characterise prevalence of dominant potentially pathogenic organisms and host immune responses.
METHODS:Sputum and nasal lavage samples were analysed using long-read metagenomic sequencing with Nanopore and qPCR in two cross-sectional adult severe asthma cohorts, Wessex (n = 66) and Oxford (n = 30). We integrated species-level data with clinical parameters and 39 selected airway proteins measured by immunoassay and O-link.
RESULTS:The sputum microbiome in health and mild asthma displayed comparable microbial diversity. By contrast, 23% (19/81) of severe asthma microbiomes were dominated by a single respiratory pathogen, namely H. influenzae (n = 10), M. catarrhalis (n = 4), S. pneumoniae (n = 4) and P. aeruginosa (n = 1). Neutrophilic asthma was associated with H. influenzae, M. catarrhalis, S. pneumoniae and T. whipplei with elevated type-1 cytokines and proteases; eosinophilic asthma with higher M. catarrhalis, but lower H. influenzae, and S. pneumoniae abundance. H. influenzae load correlated with Eosinophil Cationic Protein, elastase and IL-10. R. mucilaginosa associated positively with IL-6 and negatively with FGF. Bayesian network analysis also revealed close and distinct relationships of H. influenzae and M. catarrhalis with type-1 airway inflammation. The microbiomes and cytokine milieu were distinct between upper and lower airways.
CONCLUSION:This species-level integrated analysis reveals central, but distinct associations between potentially pathogenic bacteria and airways inflammation in severe asthma.
上一篇：
探索淋巴细胞的CD26-/lo亚群在哮喘表型和严重程度中的作用：一种表达原型粒细胞蛋白的新型CD4+T细胞亚群
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重症嗜酸性粒细胞性哮喘或嗜酸性肉芽肿性多血管炎：为新诊断策略探索潜在生物标志物
