目的:声门下狭窄(SGS)最近的转化科学努力支持一种疾病模型,其中上皮改变促进微生物组置换,失调的免疫激活,和局部纤维化。鉴于SGS中观察到的免疫细胞浸润,我们试图检验SGS病例与健康对照组相比具有低多样性(高度克隆)适应性免疫应答的假设.
方法:iSGS声门下粘膜瘢痕的单细胞RNA测序(scRNA-seq)(n=24),iLTS(n=8),进行健康对照(n=7)。提取T细胞受体(TCR)序列,分析,用来构建剧目结构,比较多样性,审讯重叠,并使用Immunarch生物信息学管道定义抗原靶标。
结果:通过Hill框架定量,健康和疾病中的近端气道粘膜具有同等差异(iSGS与iLTSvs.Control,p>0.05)。个人之间没有明显重叠(Morisita<0.02)。在iSGS患者中,TCR库的克隆性是由CD8+T细胞驱动的,和iSGS患者具有许多靶向病毒和细胞间病原体的TCR。高频克隆型不会映射到公共数据集中的已知靶标。
结论:与健康对照相比,SGS病例不具有较低多样性的适应性免疫浸润。有趣的是,健康和疾病中的TCR库都包含有限数量的高频克隆型,这些克隆型在个体之间没有显着重叠。健康和疾病中的高频克隆型的目标仍未解决。
方法:NA级喉镜,2023年。
OBJECTIVE: Recent translational scientific efforts in subglottic stenosis (SGS) support a disease model where epithelial alterations facilitate microbiome displacement, dysregulated immune activation, and localized fibrosis. Given the observed immune cell infiltrate in SGS, we sought to test the hypothesis that SGS cases possessed a low diversity (highly clonal) adaptive immune response when compared with healthy controls.
METHODS: Single cell RNA sequencing (scRNA-seq) of subglottic mucosal scar in iSGS (n = 24), iLTS (n = 8), and healthy controls (n = 7) was performed. T cell receptor (TCR) sequences were extracted, analyzed, and used to construct repertoire structure, compare diversity, interrogate overlap, and define antigenic targets using the Immunarch bioinformatics pipeline.
RESULTS: The proximal airway mucosa in health and disease are equally diverse via Hill framework quantitation (iSGS vs. iLTS vs. Control, p > 0.05). Repertoires do not significantly overlap between individuals (Morisita <0.02). Among iSGS patients, clonality of the TCR repertoire is driven by CD8+ T cells, and iSGS patients possess numerous TCRs targeting viral and intercellular pathogens. High frequency clonotypes do not map to known targets in public datasets.
CONCLUSIONS: SGS cases do not possess a lower diversity adaptive immune infiltrate when compared with healthy controls. Interestingly, the TCR repertoire in both health and disease contains a restricted number of high frequency clonotypes that do not significantly overlap between individuals. The target of the high frequency clonotypes in health and disease remain unresolved.
METHODS: NA Laryngoscope, 134:1757-1764, 2024.