Zheng Yuanzheng, Zhou Yan, Zhu Di, Fu Xing, Xie Cao, Sun Shuna, Qin Guoyou, Feng Mei, Liu Chenglong, Zhou Qingtong, Liu Fang, Chu Chen, Wang Feng, Yang Dehua, Wang Ming-Wei, Gui Yonghao
Children's Hospital of Fudan University, National Children's Medical Center, Shanghai, 201102, China.
The National Center for Drug Screening, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai, 201203, China.
Heliyon. 2024 Sep 12;10(18):e37857. doi: 10.1016/j.heliyon.2024.e37857. eCollection 2024 Sep 30.
Intravenous immunoglobulin (IVIG)-resistant Kawasaki disease (KD) poses a considerable challenge to patients and their families due to its severe complications. Previous researches have highlighted the critical role of immune disorders in its pathogenesis. However, fragmented studies based on isolated cases hinder a comprehensive understanding of this deadly illness. This study aimed to explore the overall landscape of peripheral blood mononuclear cells (PBMCs) in IVIG-resistant KD patients using single-cell RNA sequencing (scRNA-seq).
The scRNA-seq was used to characterize the transcriptomic profiles of IVIG-resistant KD patients, IVIG-responsive KD patients, and healthy controls. Data quality control (QC) and subsequent analysis were conducted using various R packages. These included DoubletFinder and Harmony for QC, Seurat and SingleR for identifying and annotating major cell types, ggpubr for calculating and visualizing the percentages of each cell type, Seurat for characterizing differentially expressed genes (DEGs) between groups, pheatmap for visualizing the DEGs, clusterProfiler for performing Gene Ontology (GO) enrichment analysis of DEGs, scRepertoire for TCR and BCR data analysis, Monocle for assessing cell differentiation trajectories, and CellChat for intercellular interaction evaluation.
High-quality single-cell transcriptome data from 12 participants were analyzed, including five with IVIG-resistant KD, four with IVIG-responsive KD, and three healthy controls. We identified 10 major cell types and observed that the differentiation of CD8 effector T cells was impeded in IVIG-resistant KD patients with coronary artery lesion (CAL) according to cell differentiation trajectory analysis. Subsequent cell communication analysis demonstrated that myeloid cluster with high expression of , , and played a key role, potentially signaling through MIF-CD74/CXCR4 and MIF-CD74/CD44 ligand-receptor pairs.
Complex immunopathological changes occur during the development of CAL in IVIG-resistant KD. Stunted differentiation of CD8 effector T cells is noted in KD-CAL. Interactions between myeloid cells and T cells activates multiple inflammatory signaling pathways, with ligand-receptor pairs, including MIF-CD74/CXCR4 and MIF-CD74/CD44, potentially playing crucial roles.
静脉注射免疫球蛋白（IVIG）抵抗的川崎病（KD）由于其严重并发症，给患者及其家庭带来了巨大挑战。以往研究强调了免疫紊乱在其发病机制中的关键作用。然而，基于个别病例的零散研究阻碍了对这种致命疾病的全面理解。本研究旨在使用单细胞RNA测序（scRNA-seq）探索IVIG抵抗的KD患者外周血单个核细胞（PBMC）的整体情况。
使用scRNA-seq对IVIG抵抗的KD患者、IVIG反应性KD患者和健康对照的转录组图谱进行表征。使用各种R包进行数据质量控制（QC）和后续分析。这些包括用于QC的DoubletFinder和Harmony，用于识别和注释主要细胞类型的Seurat和SingleR，用于计算和可视化每种细胞类型百分比的ggpubr，用于表征组间差异表达基因（DEG）的Seurat，用于可视化DEG的pheatmap，用于对DEG进行基因本体（GO）富集分析的clusterProfiler，用于TCR和BCR数据分析的scRepertoire，用于评估细胞分化轨迹的Monocle，以及用于细胞间相互作用评估的CellChat。
分析了来自12名参与者的高质量单细胞转录组数据，包括5名IVIG抵抗的KD患者、4名IVIG反应性KD患者和3名健康对照。我们识别出10种主要细胞类型，并根据细胞分化轨迹分析观察到，在患有冠状动脉病变（CAL）的IVIG抵抗的KD患者中，CD8效应T细胞的分化受到阻碍。随后的细胞通讯分析表明，高表达 、 和 的髓系簇发挥了关键作用，可能通过MIF-CD74/CXCR4和MIF-CD74/CD44配体-受体对发出信号。
IVIG抵抗的KD患者发生CAL的过程中会出现复杂的免疫病理变化。在KD-CAL中，CD8效应T细胞的分化受阻。髓系细胞与T细胞之间的相互作用激活了多个炎症信号通路，包括MIF-CD74/CXCR4和MIF-CD74/CD44在内的配体-受体对可能发挥关键作用。
