背景:过敏性真菌性鼻窦炎(AFRS)是慢性鼻窦炎伴鼻息肉病(CRSwNP)的严重表型,以局部和夸大的2型炎症为特征。虽然Th2介导的炎症的真菌抗原刺激是核心病理生理机制,细菌在疾病改变中的直接和协同作用是一个普遍的假设。我们着手定义AFRS的微环境,以阐明可能与AFRS的病理生理学有关的有毒生物。
方法:我们对AFRS患者和非真菌性CRSwNP患者进行了一项横断面研究。人口统计,疾病严重程度,对培养物和微生物组序列进行了分析。多模态微生物组测序包括IlluminaMiseq(16SrRNA和ITS)上的短读下一代测序(NGS)和牛津纳米孔技术网格(ONT)上的全长16SrRNA测序。
结果:本研究包括32例AFRS和29例非真菌性CRSwNP患者(NF)。金黄色葡萄球菌是AFRS和NF组培养和测序的优势生物(AFRS27.54%;NF18.04%;p=.07)。肺炎链球菌(AFRS12.31%;NF0.98%;p=.03)和流感嗜血杆菌(AFRS15.03%;NF0.24%;p=.005)在AFRS中明显更丰富。AFRS中的细菌多样性(香农指数)相对于NF(AFRS0.6;NF1.0,p=.008)明显较低。曲霉菌是AFRS中培养最多的真菌(10/32,31.3%)。AFRS测序的分枝杆菌组主要由马拉色菌(43.6%)代表,弯孢菌(18.5%)和曲霉菌(16.8%),而NF真菌生物群几乎完全是马拉色菌(84.2%),没有曲霉属或脱脂性真菌。
结论:多样性低,以金黄色葡萄球菌为主的微环境,肺炎链球菌和流感嗜血杆菌表征AFRS的细菌微生物组,在马拉色菌中有丰富的真菌生物群落,曲霉菌和弯孢菌。虽然金黄色葡萄球菌以前已经通过肠毒素超抗原潜力参与AFRS,肺炎链球菌和流感嗜血杆菌是新发现,可能代表替代的跨王国病理生理机制。
BACKGROUND: Allergic fungal rhinosinusitis (AFRS) is a severe phenotype of chronic rhinosinusitis with nasal polyposis (CRSwNP), characterised by localised and exaggerated type 2 inflammation. While fungal antigenic stimulation of unregulated Th2-mediated inflammation is the core pathophysiological mechanism, the direct and synergistic role of bacteria in disease modification is a pervasive hypothesis. We set out to define the microenvironment of AFRS to elucidate virulent organisms that may be implicated in the pathophysiology of AFRS.
METHODS: We undertook a cross-sectional study of AFRS patients and non-fungal CRSwNP patients. Demographics, disease severity, culture and microbiome sequences were analysed. Multimodality microbiome sequencing included short-read next-generation sequencing (NGS) on the Illumina Miseq (16S rRNA and ITS) and full-length 16S rRNA sequencing on the Oxford Nanopore Technologies GridION (ONT).
RESULTS: Thirty-two AFRS and 29 non-fungal CRSwNP patients (NF) were included in this study. Staphylococcus aureus was the dominant organism cultured and sequenced in both AFRS and NF groups (AFRS 27.54%; NF 18.04%; p = .07). Streptococcus pneumoniae (AFRS 12.31%; NF 0.98%; p = .03) and Haemophilus influenzae (AFRS 15.03%; NF 0.24%; p = .005) were significantly more abundant in AFRS. Bacterial diversity (Shannon\'s index) was considerably lower in AFRS relative to NF (AFRS 0.6; NF 1.0, p = .008). Aspergillus was the most cultured fungus in AFRS (10/32, 31.3%). The AFRS sequenced mycobiome was predominantly represented by Malassezia (43.6%), Curvularia (18.5%) and Aspergillus (16.8%), while the NF mycobiome was nearly exclusively Malassezia (84.2%) with an absence of Aspergillus or dematiaceous fungi.
CONCLUSIONS: A low diversity, dysbiotic microenvironment dominated by Staphylococcus aureus, Streptococcus pneumoniae and Haemophilus influenzae characterised the bacterial microbiome of AFRS, with a mycobiome abundant in Malassezia, Aspergillus and Curvularia. While Staphylococcus aureus has been previously implicated in AFRS through enterotoxin superantigen potential, Streptococcus pneumoniae and Haemophilus influenzae are novel findings that may represent alternate cross-kingdom pathophysiological mechanisms.