A multi-omics approach incorporating single-cell RNASequencing reveals murine Leishmania major infection drives a proinflammatory signature alongside a signature of decreased ribosomal biogenesis.

RNA-seq analysis was performed in ear samples from naïve control and L. major infected mice. Using scRNASeq, we investigated the transcriptomic changes within individual cell types present in leishmanial lesion. Transcriptomic analysis through scRNASeq revealed 35 distinct cell populations which include both resident and recruited cell types following L. major infection. In addition, our scRNASeq analysis confirms a significant recruitment of various immune cell types such as macrophages, inflammtory monocytes, neutrophils, dendritic cells, CD4 and CD8 T cells in the infected ears that are seen at higher frequencies compared to naive controls. In overall, our scRNASeq data revealed that DEGs within each immune cell type driving a proinflammtory state during L. major infection, alongside a signature of decreased ribosomal biogenesis. Overall design: Single-cell RNASeq of naïve control ears compared to ear samples from 4 week post infection with L. major promastigote parasites.

本研究针对未感染对照组（naïve control）及利什曼原虫（Leishmania major, L. major）感染小鼠的耳部样本开展了RNA测序分析。通过单细胞RNA测序（single-cell RNA sequencing, scRNA-seq），本研究探究了利什曼病皮损中各类细胞的转录组变化。本次单细胞RNA测序的转录组分析共鉴定出35种不同的细胞群，涵盖利什曼原虫感染后存在的固有驻留细胞与招募浸润细胞两类。此外，本研究的单细胞RNA测序分析证实，与未感染对照组相比，感染小鼠耳部中各类免疫细胞的招募显著增多，包括巨噬细胞、促炎单核细胞、中性粒细胞、树突状细胞、CD4⁺和CD8⁺ T细胞，且这些免疫细胞的占比更高。总体而言，本研究的单细胞RNA测序数据显示，在利什曼原虫感染过程中，各类免疫细胞内的差异表达基因（differentially expressed genes, DEGs）驱动了促炎状态的形成，同时伴随核糖体生物发生水平下调的特征。实验整体设计：将未感染对照组小鼠耳部样本，与感染利什曼原虫前鞭毛体4周后的小鼠耳部样本进行单细胞RNA测序对比分析。