Abstract：
Rationale: Fibrotic hypersensitivity pneumonitis is a debilitating interstitial lung disease driven by incompletely understood immune mechanisms. Objectives: To elucidate immune aberrations in fibrotic hypersensitivity pneumonitis in single-cell resolution. Methods: Single-cell 5\' RNA sequencing was conducted on peripheral blood mononuclear cells and bronchoalveolar lavage cells obtained from 45 patients with fibrotic hypersensitivity pneumonitis, 63 idiopathic pulmonary fibrosis, 4 non-fibrotic hypersensitivity pneumonitis, and 36 healthy controls in the United States and Mexico. Analyses included differential gene expression (Seurat), transcription factor activity imputation (DoRothEA-VIPER), and trajectory analyses (Monocle3/Velocyto-scVelo-CellRank). Measurements and Main Results: Overall, 501,534 peripheral blood mononuclear cells from 110 patients and controls and 88,336 bronchoalveolar lavage cells from 19 patients were profiled. Compared to controls, fibrotic hypersensitivity pneumonitis has elevated classical monocytes (adjusted-p=2.5e-3) and are enriched in CCL3hi/CCL4hi and S100Ahi classical monocytes (adjusted-p<2.2e-16). Trajectory analyses demonstrate that S100Ahi classical monocytes differentiate into SPP1hi lung macrophages associated with fibrosis. Compared to both controls and idiopathic pulmonary fibrosis, fibrotic hypersensitivity pneumonitis patient cells are significantly enriched in GZMhi cytotoxic T cells. These cells exhibit transcription factor activities indicative of TGFβ and TNFα/NFκB pathways. These results are publicly available at https://ildimmunecellatlas.org. Conclusions: Single-cell transcriptomics of fibrotic hypersensitivity pneumonitis patients uncovered novel immune perturbations, including previously undescribed increases in GZMhi cytotoxic CD4+ and CD8+ T cells - reflecting this disease\'s unique inflammatory T-cell driven nature - as well as increased S100Ahi and CCL3hi/CCL4hi classical monocytes also observed in idiopathic pulmonary fibrosis. Both cell populations may guide the development of new biomarkers and therapeutic interventions.
摘要：
原理：纤维性过敏性肺炎是一种由不完全了解的免疫机制驱动的衰弱性间质性肺病。目的：以单细胞分辨率阐明纤维化过敏性肺炎的免疫异常。方法：对45例纤维化过敏性肺炎患者的外周血单个核细胞和支气管肺泡灌洗液细胞进行单细胞5'RNA测序，63特发性肺纤维化，4非纤维化过敏性肺炎，以及美国和墨西哥的36名健康对照。分析包括差异基因表达（Seurat），转录因子活性插补(DoRothEA-VIPER)，和轨迹分析(Monocle3/Velocyto-scVelo-CellRank)。测量和主要结果：总体，对110例患者和对照组的501,534个外周血单个核细胞和19例患者的88,336个支气管肺泡灌洗细胞进行了分析。与对照组相比，纤维化过敏性肺炎具有升高的经典单核细胞(调整-p=2.5e-3),并且富含CCL3hi/CCL4hi和S100Ahi经典单核细胞(调整-p<2.2e-16)。轨迹分析表明，S100Ahi经典单核细胞分化成与纤维化相关的SPP1hi肺巨噬细胞。与对照组和特发性肺纤维化相比，纤维化过敏性肺炎患者细胞在GZMhi细胞毒性T细胞中显著富集。这些细胞表现出指示TGFβ和TNFα/NFκB途径的转录因子活性。这些结果可在https://ildimunecellatlas.org上公开获得。结论：纤维化过敏性肺炎患者的单细胞转录组学发现了新的免疫扰动，包括先前未描述的GZMhi细胞毒性CD4+和CD8+T细胞的增加-反映了该疾病独特的炎性T细胞驱动性质-以及在特发性肺纤维化中也观察到的S100Ahi和CCL3hi/CCL4hi经典单核细胞的增加。两种细胞群都可以指导新的生物标志物和治疗干预措施的开发。
