目的:提出特发性声门下狭窄(iSGS)的综合流式细胞术小组。
方法:对照离体队列研究。
方法:大都市地区的三级护理学术医院。
方法:对iSGS患者的9个配对的正常和瘢痕组织样本进行流式细胞术和单细胞RNA测序。流式细胞术用于评估髓样的存在(CD11b,CD14,CD15,Siglec8),淋巴样(CD3,CD4,CD8,γδ[γδ],FOXP3),内皮(CD31),成纤维细胞(CD90,SMA),和上皮(CD326,CK5)标记。
结果:关于流式细胞术,iSGS疤痕的特点是骨髓的存在增加,淋巴样,内皮,和成纤维细胞类型,而是减少了上皮细胞的存在。在骨髓谱系中,iSGS疤痕样本显示CD11b+单核细胞增加(P<.001),Siglec8+嗜酸性粒细胞(P=0.03),和CD14+单核细胞(P=.02)。在淋巴谱系中,iSGS瘢痕显示CD3+T细胞增加(P<.001),CD4+辅助性T细胞(P<0.001),γδ+T细胞(P<.001),和FOXP3+调节性T细胞(P=0.002)。iSGS瘢痕表现出CD90+(P=.04)和SMA+(P<.001)成纤维细胞的特异性增加,但CD326+(E-cadherin)上皮细胞相对于正常样品减少(P=.01)。
结论:我们为iSGS提供了一个全面的流式细胞术小组。该流程面板可以作为气道科学家之间的共同平台,以阐明支撑iSGS和其他上呼吸道病理的细胞机制。疤痕iSGS样本相对于正常iSGS样本显示出独特的细胞特征,表现出增加的成纤维细胞,内皮,和炎症细胞类型,但上皮减少。
OBJECTIVE: To present a comprehensive flow cytometry panel for idiopathic subglottic stenosis (iSGS).
METHODS: Controlled ex vivo cohort study.
METHODS: Tertiary care academic hospital in a metropolitan area.
METHODS: Flow cytometry and single-cell RNA sequencing were performed on 9 paired normal and scar tissue samples from iSGS patients. Flow cytometry was used to assess the presence of myeloid (CD11b, CD14, CD15, Siglec8), lymphoid (CD3, CD4, CD8, gamma delta [γδ], FOXP3), endothelial (CD31), fibroblast (CD90, SMA), and epithelial (CD326, CK5) markers.
RESULTS: On flow cytometry, iSGS scar is characterized by an increased presence of myeloid, lymphoid, endothelial, and fibroblast cell types, but a decreased presence of epithelial cells. In the myeloid lineage, iSGS scar samples demonstrated increased CD11b+ monocytes (P < .001), Siglec8+ eosinophils (P = .03), and CD14+ monocytes (P = .02). In the lymphoid lineage, iSGS scar demonstrated increased CD3+ T-cells (P < .001), CD4+ helper T-cells (P < .001), γδ+ T-cells (P < .001), and FOXP3+ regulatory T-cells (P = .002). iSGS scar exhibited specific increases in CD90+ (P = .04) and SMA+ (P < .001) fibroblasts but decreased CD326+ (E-cadherin) epithelial cells (P = .01) relative to normal samples.
CONCLUSIONS: We present a comprehensive flow cytometry panel for iSGS. This flow panel may serve as a common platform among airway scientists to elucidate the cellular mechanisms underpinning iSGS and other upper airway pathologies. Scar iSGS samples demonstrate a distinct cellular profile relative to normal iSGS specimens, exhibiting increased fibroblast, endothelial, and inflammatory cell types but decreased epithelium.