Department of Otolaryngology-Head and Neck Surgery, Johns Hopkins University School of Medicine, Baltimore, Maryland, USA.
Bloomberg~Kimmel Institute for Cancer Immunotherapy, Baltimore, Maryland, USA.
Otolaryngol Head Neck Surg. 2021 Jun;164(6):1257-1264. doi: 10.1177/0194599820978271. Epub 2020 Dec 8.
Iatrogenic laryngotracheal stenosis (iLTS) is characterized by fibroinflammatory narrowing of the upper airway and is most commonly caused by intubation injury. Evidence suggests a key role for CD4 T cells in its pathogenesis. The objective of this study is to validate emerging multiplex immunofluorescence (mIF) technology for use in the larynx and trachea while quantitatively characterizing the immune cell infiltrate in iLTS. In addition to analyzing previously unstudied immune cell subsets, this study aims to validate previously observed elevations in the immune checkpoint PD-1 and its ligand PD-L1 while exploring their spatial and cellular distributions in the iLTS microenvironment.
Controlled ex vivo cohort study.
Tertiary care center.
mIF staining was performed with formalin-fixed, paraffin-embedded slides from 10 patients with iLTS who underwent cricotracheal resection and 10 control specimens derived from rapid autopsy for CD4, CD8, CD20, FoxP3, PD-1, PD-L1, and cytokeratin.
There was greater infiltration of CD4 T cells, CD8 T cells, CD20 B cells, FoxP3CD4 Tregs, and FoxP3CD8 early effector T cells in the submucosa of iLTS specimens as compared with controls ( < .05 for all). PD-1 was primarily expressed on T cells and PD-L1 predominantly on CD4 cells and "other" cells.
This study leverages the power of mIF to quantify the iLTS immune infiltrate in greater detail. It confirms the highly inflammatory nature of iLTS, with CD4 cells dominating the immune cell infiltrate; it further characterizes the cellular and spatial distribution of PD-1 and PD-L1; and it identifies novel immunologic targets in iLTS.
医源性喉气管狭窄（iLTS）的特征是上呼吸道纤维炎性狭窄，最常见的原因是插管损伤。有证据表明 CD4 T 细胞在其发病机制中起关键作用。本研究的目的是验证新兴的多重免疫荧光（mIF）技术在喉和气管中的应用，同时定量描述 iLTS 中的免疫细胞浸润。除了分析以前未研究过的免疫细胞亚群外，本研究旨在验证以前观察到的免疫检查点 PD-1 及其配体 PD-L1 的升高，同时探索它们在 iLTS 微环境中的空间和细胞分布。
对照性离体队列研究。
三级保健中心。
用 10 例接受环气管切开术的 iLTS 患者的福尔马林固定、石蜡包埋切片和 10 例快速尸检对照标本进行 mIF 染色，检测 CD4、CD8、CD20、FoxP3、PD-1、PD-L1 和细胞角蛋白。
与对照组相比，iLTS 标本的黏膜下有更多的 CD4 T 细胞、CD8 T 细胞、CD20 B 细胞、FoxP3CD4 Treg 和 FoxP3CD8 早期效应 T 细胞浸润（所有 P 值均 <.05）。PD-1 主要在 T 细胞上表达，PD-L1 主要在 CD4 细胞和“其他”细胞上表达。
本研究利用 mIF 的力量更详细地量化 iLTS 的免疫浸润。它证实了 iLTS 的高度炎症性质，CD4 细胞主导免疫细胞浸润；它进一步描述了 PD-1 和 PD-L1 的细胞和空间分布；并确定了 iLTS 中的新免疫靶标。
