严重哮喘中聚肌苷酸胞苷酸刺激外周血单个核细胞模型的单细胞特征分析。
Single-cell characterization of a model of poly I:C-stimulated peripheral blood mononuclear cells in severe asthma.
机构信息
Section of Pulmonary, Critical Care and Sleep Medicine, Department of Internal Medicine, Yale University School of Medicine, 300 Cedar Street (S419 TAC), New Haven, CT, 06520-8057, USA.
Department of Oncology, Clínica Universidad de Navarra, Pamplona, Spain.
出版信息
Respir Res. 2021 Apr 26;22(1):122. doi: 10.1186/s12931-021-01709-9.
BACKGROUND
Asthma has been associated with impaired interferon response. Multiple cell types have been implicated in such response impairment and may be responsible for asthma immunopathology. However, existing models to study the immune response in asthma are limited by bulk profiling of cells. Our objective was to Characterize a model of peripheral blood mononuclear cells (PBMCs) of patients with severe asthma (SA) and its response to the TLR3 agonist Poly I:C using two single-cell methods.
METHODS
Two complementary single-cell methods, DropSeq for single-cell RNA sequencing (scRNA-Seq) and mass cytometry (CyTOF), were used to profile PBMCs of SA patients and healthy controls (HC). Poly I:C-stimulated and unstimulated cells were analyzed in this study.
RESULTS
PBMCs (n = 9414) from five SA (n = 6099) and three HC (n = 3315) were profiled using scRNA-Seq. Six main cell subsets, namely CD4 + T cells, CD8 + T cells, natural killer (NK) cells, B cells, dendritic cells (DCs), and monocytes, were identified. CD4 + T cells were the main cell type in SA and demonstrated a pro-inflammatory profile characterized by increased JAK1 expression. Following Poly I:C stimulation, PBMCs from SA had a robust induction of interferon pathways compared with HC. CyTOF profiling of Poly I:C stimulated and unstimulated PBMCs (n = 160,000) from the same individuals (SA = 5; HC = 3) demonstrated higher CD8 + and CD8 + effector T cells in SA at baseline, followed by a decrease of CD8 + effector T cells after poly I:C stimulation.
CONCLUSIONS
Single-cell profiling of an in vitro model using PBMCs in patients with SA identified activation of pro-inflammatory pathways at baseline and strong response to Poly I:C, as well as quantitative changes in CD8 + effector cells. Thus, transcriptomic and cell quantitative changes are associated with immune cell heterogeneity in this model to evaluate interferon responses in severe asthma.
背景
哮喘与干扰素反应受损有关。多种细胞类型都与这种反应受损有关，可能是哮喘免疫病理学的原因。然而，现有的研究哮喘免疫反应的模型受到细胞批量分析的限制。我们的目的是使用两种单细胞方法来描述严重哮喘（SA）患者的外周血单核细胞（PBMC）模型及其对 TLR3 激动剂 Poly I:C 的反应。
方法
使用两种互补的单细胞方法，即 DropSeq 用于单细胞 RNA 测序（scRNA-Seq）和质谱流式细胞术（CyTOF），对 SA 患者和健康对照（HC）的 PBMC 进行分析。本研究分析了 Poly I:C 刺激和未刺激的细胞。
结果
使用 scRNA-Seq 对来自五名 SA（n=6099）和三名 HC（n=3315）的 9414 个 PBMC 进行了分析。鉴定了六个主要的细胞亚群，即 CD4+T 细胞、CD8+T 细胞、自然杀伤（NK）细胞、B 细胞、树突状细胞（DC）和单核细胞。CD4+T 细胞是 SA 的主要细胞类型，表现出以 JAK1 表达增加为特征的促炎表型。在 Poly I:C 刺激后，与 HC 相比，SA 的 PBMC 中干扰素途径有强烈的诱导。来自同一个体（SA=5；HC=3）的 Poly I:C 刺激和未刺激 PBMC（n=160000）的 CyTOF 分析显示，SA 患者在基线时有更高的 CD8+和 CD8+效应 T 细胞，随后 Poly I:C 刺激后 CD8+效应 T 细胞减少。
结论
使用 SA 患者的 PBMC 对体外模型进行单细胞分析，在基线时鉴定出促炎途径的激活和对 Poly I:C 的强烈反应，以及 CD8+效应细胞的定量变化。因此，转录组和细胞定量变化与该模型中的免疫细胞异质性相关，可用于评估严重哮喘中的干扰素反应。
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