背景:嗜酸性粒细胞是与过敏性炎症有关的难以捉摸的细胞。单细胞RNA测序(scRNA-seq)是一种深入表征细胞特性的新兴方法。异质性,和功能。
目的:全面表征人类嗜酸性粒细胞在食道严重过敏性炎症部位的转录组和生物学功能(即,嗜酸性粒细胞性食管炎(EoE))。
方法:我们采用基于重力的scRNA-seq方法学对来自EoE患者和对照个体的血液嗜酸性粒细胞进行测序,与重新分析的EoE患者食管嗜酸性粒细胞的公开scRNA-seq数据集进行比较。我们用流式细胞仪,免疫染色,和刺激试验以验证mRNA的发现。
结果:总计,scRNA-seq来自586个嗜酸性粒细胞(188个来自血液[n=6个个体]和398个来自食道[n=6个个体])。食管嗜酸性粒细胞由激活的嗜酸性粒细胞群(与外周血相关嗜酸性粒细胞相比富含659个基因)和少量类似外周血嗜酸性粒细胞的嗜酸性粒细胞群(与食管嗜酸性粒细胞相比富含62个基因)组成。食管嗜酸性粒细胞表达的基因参与感知和响应不同的刺激,最值得注意的是干扰素-,白细胞介素10,组胺和白三烯,和琥珀酸。食管嗜酸性粒细胞与其他食管人群的区别在于受体CCR3,HRH4,SUCNR1和VSTM1的基因表达;转录因子CEBPE,OLIG1和OLIG2;蛋白酶PRSS33;和标志嗜酸性粒细胞基因CLC。得出了与其他食道人群的双向嗜酸性粒细胞相互作用的网络。比较食管嗜酸性粒细胞和肥大细胞显示食管嗜酸性粒细胞表达参与DAP12相互作用的基因,IgG受体触发事件,免疫调节,和IL-10信号。
结论:在EoE中,食管嗜酸性粒细胞存在两个种群,一个类似于血液嗜酸性粒细胞的少数群体,另一个群体的特征是不同的感应受体和炎症介质的高从头转录,使它们可能与不同的细胞类型相交。
BACKGROUND: Eosinophils are elusive cells involved in allergic inflammation. Single-cell RNA-sequencing (scRNA-seq) is an emerging approach to deeply characterize cellular properties, heterogeneity, and functionality.
OBJECTIVE: We sought to comprehensively characterize the transcriptome and biological functions of human eosinophils at a site of severe allergic inflammation in the esophagus (ie, eosinophilic esophagitis [EoE]).
METHODS: We employed a gravity-based scRNA-seq methodology to sequence blood eosinophils from patients with EoE and control individuals compared to a reanalyzed public scRNA-seq dataset of human esophageal eosinophils of EoE patients. We used flow cytometry, immunostaining, and a stimulation assay to verify mRNA findings.
RESULTS: In total, scRNA-seq was obtained from 586 eosinophils (188 from blood [n = 6 individuals] and 398 from esophagus [n = 6 individuals]). The esophageal eosinophils were composed of a population of activated eosinophils (enriched in 659 genes compared with peripheral blood-associated eosinophils) and a small population of eosinophils resembling peripheral blood eosinophils (enriched in 62 genes compared with esophageal eosinophils). Esophageal eosinophils expressed genes involved in sensing and responding to diverse stimuli, most notably IFN-γ, IL-10, histamine and leukotrienes, and succinate. Esophageal eosinophils were most distinguished from other esophageal populations by gene expression of the receptors
CCR3, HRH4, SUCNR1, and VSTM1; transcription factors CEBPE, OLIG1, and OLIG2; protease PRSS33; and the hallmark eosinophil gene CLC. A web of bidirectional eosinophil interactions with other esophageal populations was derived. Comparing esophageal eosinophils and mast cells revealed that esophageal eosinophils expressed genes involved in DNAX-activation protein-12 (also known as TYROBP) interactions, IgG receptor-triggered events, immunoregulation, and IL-10 signaling.
CONCLUSIONS: In EoE, esophageal eosinophils exist as 2 populations, a minority population resembling blood eosinophils and the other population characterized by high de novo transcription of diverse sensing receptors and inflammatory mediators readying them to potentially intersect with diverse cell types.