Single-cell RNA-sequencing of human cytomegalovirus-infected humanized mice

Human cytomegalovirus (HCMV) is a common herpesvirus that persistently infects a large portion of the world's population. Despite the robust host immune response, HCMV is able to replicate, evade host defenses, and establish latency throughout the lifespan by developing multiple immunomodulatory strategies, making the studies on the interaction between HCMV infection and host response particularly important. HCMV has a strict host specificity that specifically infects humans. Therefore, most of the in vivo researches of HCMV rely on clinical samples. Fortunately, the establishment of humanized mouse models allows for convenient in-lab animal experiments involving HCMV infection. Single-cell RNA sequencing(scRNA-seq) enables the study of the relationship between viral and host gene expressions at the single-cell level within host cells. In this study, we assessed the gene expression alterations of PBMCs at the single-cell level within HCMV-infected humanized mice, which sheds light onto the virus-host interactions in the context of HCMV infecton of humanized mice and provides a valuable dataset for the related researches. To delineate the single-cell dynamics of peripheral blood mononuclear cells (PBMCs) during latent and reactivated HCMV infection, we procured PBMCs from the peripheral blood of humanized mice. These were categorized into two groups: two wild-type (Mock), and two HCMV-infected (HCMV). Initially, NHDF cells harboring HCMV were introduced into the peritoneal cavity of NOD-scid IL2Rgcnull mice. Following a 4-week period peripheral blood was harvested and partitioned from each mouse group

人类巨细胞病毒（Human cytomegalovirus, HCMV）是一种常见疱疹病毒，可在全球绝大多数人群中建立持续性感染。尽管宿主可产生强大的免疫应答，HCMV仍可通过演化出多种免疫调节策略完成复制、逃避宿主防御并在宿主终生维持潜伏感染，因此研究HCMV感染与宿主应答之间的相互作用具有重要意义。HCMV具有严格的宿主特异性，仅感染人类，因此绝大多数体内研究依赖临床样本。值得庆幸的是，人源化小鼠模型的建立使得实验室中可便捷开展涉及HCMV感染的动物实验。单细胞RNA测序（single-cell RNA sequencing, scRNA-seq）可实现在单细胞层面解析宿主细胞内病毒与宿主基因表达的关联。本研究对HCMV感染的人源化小鼠体内的外周血单个核细胞（peripheral blood mononuclear cells, PBMCs）进行了单细胞水平的基因表达变化分析，为阐明人源化小鼠HCMV感染情境下的病毒-宿主相互作用提供了关键见解，同时为相关研究提供了宝贵的数据集。为解析潜伏感染与再激活HCMV感染过程中外周血单个核细胞的单细胞动态变化，我们从人源化小鼠的外周血中采集了PBMCs，并将其分为两组：两组野生型（Mock，空白对照）与两组HCMV感染组。实验初始，我们将携带HCMV的正常人皮肤成纤维细胞（NHDF）接种至NOD-scid IL2Rgcnull小鼠的腹腔内。4周后，采集各组小鼠的外周血并完成分离。