We downloaded single-cell RNA-seq expression data from GEO with accession number GSE207651. The mice underwent cecal ligation and puncture (CLP) surgery, and lung tissue samples were collected at 0, 24, and 48 h. The cells were annotated using the CellMarker database and FindAllMarkers functions. GO enrichment analyses were performed using the Metascape software. Gene set enrichment Analysis (GSEA) and variation Analysis (GSVA) were performed to identify differential signaling pathways. Differential expression genes were collected with the \"FindMarkers\" function. The R package AUCell was used to score individual cells for pathway activities. The Cellchat package was used to explore intracellular communication.
Granulocytes increased significantly as the duration of endotoxemia increased. However, the number of T cells, NK cells, and B cells declined. Pulmonary capillary cells were grouped into three sub-clusters. Capillary-3 cells were enriched in the sham group, but declined sharply in the CLP.24 group. Capillary-1 cells peaked in the CLP.24 group, while Capillary-2 cells were enriched in the CLP.48 group. Furthermore, we found that Cd74+ Capillary-3 cells mainly participated in immune interactions. Plat+ Capillary-1 and Clec1a+ Capillary-2 are involved in various physiological processes. Regarding cell-cell interactions, Plat+ Capillary-1 plays the most critical role in granulocyte adherence to capillaries during ALI. Cd74+ Capillary cells expressing high levels of major histocompatibility complex (MHC) and mainly interacted with Cd8a+ T cells in the sham group.
Plat+ capillaries are involved in the innate immune response through their interaction with neutrophils via ICAM-1 adhesion during endotoxemia, while Cd74+ capillaries epxressed high level of MHC proteins play a role in adaptive immune response through their interaction with T cells. However, it remains unclear whether the function of Cd74+ capillaries leans towards immunity or tolerance, and further studies are needed to confirm this.
■我们从GEO下载了登录号为GSE207651的单细胞RNA-seq表达数据。小鼠进行盲肠结扎和穿刺(CLP)手术,在0、24和48小时收集肺组织样品。使用CellMarker数据库和FindAllMarkers函数对细胞进行注释。使用Metascape软件进行GO富集分析。进行基因集富集分析(GSEA)和变异分析(GSVA)以鉴定差异信号通路。收集具有“FindMarkers”功能的差异表达基因。使用R包AUCell对单个细胞的途径活性进行评分。Cellchat包用于探索细胞内通信。
■粒细胞随着内毒素血症持续时间的增加而显著增加。然而,T细胞的数量,NK细胞,B细胞下降。肺毛细血管细胞分为三个亚簇。在假手术组中富集毛细管-3细胞,但在CLP.24组急剧下降。毛细管-1细胞在CLP.24组中达到峰值,而在CLP.48组中富集了毛细管-2细胞。此外,我们发现Cd74+毛细管-3细胞主要参与免疫相互作用。平板+毛细管-1和Clec1a+毛细管-2参与各种生理过程。关于细胞间的相互作用,Plat毛细管-1在ALI期间粒细胞粘附到毛细血管中起着最关键的作用。假手术组Cd74+细胞表达高水平的主要组织相容性复合体(MHC),主要与Cd8a+T细胞相互作用。
■血小板+毛细血管在内毒素血症期间通过ICAM-1粘附与中性粒细胞相互作用而参与先天免疫反应,而Cd74+毛细血管表达高水平的MHC蛋白通过与T细胞的相互作用在适应性免疫应答中发挥作用。然而,目前尚不清楚Cd74+毛细血管的功能是否倾向于免疫或耐受性,需要进一步的研究来证实这一点。