Lung myeloid cells are important in pulmonary immune homeostasis and in the pathogenesis of chronic obstructive pulmonary disease (COPD). Multiparameter immunophenotypic characterization of these cells is challenging because of their autofluorescence and diversity. We evaluated the immunophenotypic landscape of airway myeloid cells in COPD using time of flight mass cytometry. Cells from BAL, which were obtained from never-smokers (n = 8) and smokers with (n = 20) and without (n = 4) spirometric COPD, were examined using a 44-parameter time of flight mass cytometry panel. Unsupervised cluster analysis was used to identify cellular subtypes that were confirmed by manual gating. We identified major populations of CD68⁺ and CD68⁻ cells with 22 distinct phenotypic clusters, of which 18 were myeloid cells. We found a higher abundance of putative recruited myeloid cells (CD68⁺ classical monocytes) in BAL from patients with COPD. CD68⁺ classical monocyte population had distinct responses to smoking and COPD that were potentially related to their recruitment from the interstitium and vasculature. We demonstrate that BAL cells from smokers and subjects with COPD have lower AXL expression. Also, among subjects with COPD, we report significant differences in the abundance of PDL1^high and PDL2^high clusters and in the expression of PDL1 and PDL2 across several macrophage subtypes suggesting modulation of inflammatory responses. In addition, several phenotypic differences in BAL cells from subjects with history of COPD exacerbation were identified that could inform potential disease mechanisms. Overall, we report several changes to the immunophenotypic landscape that occur with smoking, COPD, and past exacerbations that are consistent with decreased regulation and increased activation of inflammatory pathways.

肺髓样细胞在肺免疫稳态和慢性阻塞性肺病 (COPD) 的发病机制中很重要。由于这些细胞的自发荧光和多样性，对这些细胞的多参数免疫表型表征具有挑战性。我们使用飞行时间质谱流式细胞术评估了 COPD 中气道骨髓细胞的免疫表型景观。来自从不吸烟者 ( n  = 8) 和吸烟者 ( n  = 20) 和没有 ( n  = 4) 肺活量 COPD 的BAL 细胞使用 44 参数飞行时间质谱仪面板进行检查。无监督聚类分析用于识别通过手动门控确认的细胞亚型。我们确定了 CD68 ⁺和 CD68 的主要群体^-具有 22 个不同表型簇的细胞，其中 18 个是骨髓细胞。我们在 COPD 患者的 BAL 中发现了更高丰度的推定募集的骨髓细胞（CD68 ⁺经典单核细胞）。CD68 ⁺经典单核细胞群对吸烟和 COPD 有不同的反应，这可能与它们从间质和脉管系统的募集有关。我们证明来自吸烟者和 COPD 受试者的 BAL 细胞具有较低的 AXL 表达。此外，在患有 COPD 的受试者中，我们报告了 PDL1^高和 PDL2^高丰度的显着差异簇和 PDL1 和 PDL2 在几种巨噬细胞亚型中的表达表明炎症反应的调节。此外，还确定了来自具有 COPD 急性加重史的受试者的 BAL 细胞的几种表型差异，可以为潜在的疾病机制提供信息。总体而言，我们报告了吸烟、COPD 和既往恶化导致的免疫表型景观的一些变化，这些变化与炎症通路的调节减少和激活增加一致。