IntroductionAllergic 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.MethodologyWe 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).ResultsThirty‐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.ConclusionA 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.

中文翻译：

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使用全长 16S rRNA 基因扩增子测序和真菌 ITS 测序表征过敏性真菌性鼻窦炎微环境


简介过敏性真菌性鼻窦炎 (AFRS) 是慢性鼻窦炎伴鼻息肉病 (CRSwNP) 的一种严重表型，其特征是局部且严重的 2 型炎症。虽然真菌抗原刺激不受调节的 Th2 介导的炎症是核心病理生理机制，但细菌在疾病缓解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 病理生理学有关的有毒微生物。方法学我们对 AFRS 患者和非真菌 CRSwNP 患者进行了横断面研究。分析了人口统计、疾病严重程度、培养和微生物组序列。多模态微生物组测序包括 Illumina Miseq（16S rRNA 和 ITS）上的短读长新一代测序 (NGS) 以及 Oxford Nanopore Technologies GridION (ONT) 上的全长 16S rRNA 测序。结果 32 个 AFRS 和 29 个非真菌本研究包括 CRSwNP 患者 (NF)。金黄色葡萄球菌是 AFRS 和 NF 组中培养和测序的优势微生物（AFRS 27.54%；NF 18.04%； p = .07)。肺炎链球菌（AFRS 12.31%；NF 0.98%； p = .03) 和流感嗜血杆菌（AFRS 15.03%；NF 0.24%； p = .005）在 AFRS 中显着更丰富。 AFRS 中的细菌多样性（香农指数）远低于 NF（AFRS 0.6；NF 1.0， p =.008）。曲霉属是 AFRS 中培养最多的真菌（10/32，31.3%）。 AFRS 测序的真菌组主要代表为马拉色菌(43.6%), 弯孢菌 (18.5%）和曲霉属(16.8%)，而 NF 真菌组几乎完全是马拉色菌(84.2%) 没有曲霉属或暗色真菌。结论金黄色葡萄球菌、肺炎链球菌和流感嗜血杆菌AFRS 的细菌微生物组特征，其中真菌组丰富马拉色菌,曲霉属和弯孢属。尽管金黄色葡萄球菌之前已通过肠毒素超抗原潜力与 AFRS 相关，肺炎链球菌和流感嗜血杆菌这些新颖的发现可能代表了不同的跨界病理生理机制。