Single-cell characterization of a model of poly I:C-stimulated peripheral blood mononuclear cells in severe asthma.
Single-cell characterization of a model of poly I:C-stimulated peripheral blood mononuclear cells in severe asthma.
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DOI：
10.1186/s12931-021-01709-9
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发表时间：
2021-04-26
影响因子：
5.8
通讯作者：
Gomez JL
中科院分区：
文献类型：
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作者：
Chen A;Diaz-Soto MP;Sanmamed MF;Adams T;Schupp JC;Gupta A;Britto C;Sauler M;Yan X;Liu Q;Nino G;Cruz CSD;Chupp GL;Gomez JL
哮喘与干扰素应答受损有关。多种类型的细胞参与了这种反应障碍，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表征严重哮喘（SA）患者外周血单核细胞（PBMC）模型及其对TLR 3激动剂Poly I：C的反应。使用两种互补的单细胞方法，即用于单细胞RNA测序的DropSeq（scRNA-Seq）和质谱细胞术（CyTOF）来分析SA患者和健康对照（HC）的PBMCs。在这项研究中分析了Poly I：C刺激和未刺激的细胞。使用scRNA-Seq对来自五个SA（n = 6099）和三个HC（n = 3315）的PBMC（n = 9414）进行分析。鉴定了六种主要的细胞亚群，即CD 4 + T细胞、CD 8 + T细胞、自然杀伤（NK）细胞、B细胞、树突状细胞（DC）和单核细胞。CD 4 + T细胞是SA中的主要细胞类型，并表现出以JAK 1表达增加为特征的促炎特征。在Poly I：C刺激后，与HC相比，来自SA的PBMC对干扰素途径具有稳健的诱导。来自相同个体（SA = 5; HC = 3）的Poly I：C刺激和未刺激PBMC（n = 160，000）的CyTOF分析表明，基线时SA中的CD 8+和CD 8+效应T细胞较高，随后在Poly I：C刺激后CD 8+效应T细胞减少。在SA患者中使用PBMC的体外模型的单细胞分析确定了基线时促炎途径的激活和对Poly I：C的强烈应答，以及CD 8+效应细胞的定量变化。因此，转录组和细胞数量的变化与免疫细胞异质性在这个模型中，以评估严重哮喘的干扰素反应。在线版本包含补充材料，可通过10.1186/s12931-021-01709-9获得。
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. 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. 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. 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. The online version contains supplementary material available at 10.1186/s12931-021-01709-9.
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影响因子：
12.3
作者：
Guillaumet-Adkins A;Rodríguez-Esteban G;Mereu E;Mendez-Lago M;Jaitin DA;Villanueva A;Vidal A;Martinez-Marti A;Felip E;Vivancos A;Keren-Shaul H;Heath S;Gut M;Amit I;Gut I;Heyn H
通讯作者：
Heyn H
影响因子：
4.3
作者：
Chen H;Lau MC;Wong MT;Newell EW;Poidinger M;Chen J
通讯作者：
Chen J
影响因子：
64.8
作者：
Alexopoulou, L;Holt, AC;Flavell, RA
通讯作者：
Flavell, RA
影响因子：
24.3
作者：
Chung, Kian Fan;Wenzel, Sally E.;Teague, W. Gerald
通讯作者：
Teague, W. Gerald
影响因子：
30.5
作者：
通讯作者：
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