Single-Cell Transcriptome Analysis of CD34+ Stem Cell-Derived Myeloid Cells Infected With Human Cytomegalovirus
Single-Cell Transcriptome Analysis of CD34+ Stem Cell-Derived Myeloid Cells Infected With Human Cytomegalovirus
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DOI：
10.3389/fmicb.2019.00577
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发表时间：
2019-03-21
影响因子：
5.2
通讯作者：
Hertel, Laura
中科院分区：
文献类型：
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作者：
Galinato, Melissa;Shimoda, Kristen;Hertel, Laura
髓系细胞是人巨细胞病毒(CMV)裂解和潜伏感染的重要部位。我们之前的研究表明，从CD34(+)造血干细胞分化而来的髓系细胞只有一小部分可以复制CMV，这突显了这些群体的异质性。然而，抗性和允许细胞类型的确切相同，以及后者的细胞特征，不能用平均转录分析工具(如微阵列)来剖析，因此仍然是个谜。在这里，我们使用10x基因组学平台描述了感染后第一天的大约7000个单个细胞的转录本。我们发现病毒转录本可以在大多数细胞中检测到，这表明病毒粒子进入不太可能是细胞限制机制的主要目标。我们进一步表明，病毒复制发生在一小部分特定的细胞亚群中，这些细胞在转录上与表达集落形成单位标志的细胞簇-粒细胞、红细胞、单核细胞、巨核细胞(CFU-GEMM)-相关，并且可能来源于这些细胞群。与其他细胞相比，CFU-GEMM细胞具有丰富的转录本，具有线粒体能量产生、细胞增殖、RNA处理和蛋白质合成的功能，并表达类似或更高水平的干扰素相关基因。虽然前者的表达水平在感染细胞中保持不变，但后者的表达水平却强烈下调。因此，我们认为CFU-GEMM细胞的优先感染可能是由于预先建立的亲病毒环境的存在，而不是由于缺乏特定的限制因素，该环境需要病毒效应器做出最小的优化努力。总之，这些发现确定了一个潜在的新的允许CMV复制的髓系细胞群体，并为其优先感染提供了可能的理论基础。
Myeloid cells are important sites of lytic and latent infection by human cytomegalovirus (CMV). We previously showed that only a small subset of myeloid cells differentiated from CD34(+) hematopoietic stem cells is permissive to CMV replication, underscoring the heterogeneous nature of these populations. The exact identity of resistant and permissive cell types, and the cellular features characterizing the latter, however, could not be dissected using averaging transcriptional analysis tools such as microarrays and, hence, remained enigmatic. Here, we profile the transcriptomes of similar to 7000 individual cells at day 1 post-infection using the 10x genomics platform. We show that viral transcripts are detectable in the majority of the cells, suggesting that virion entry is unlikely to be the main target of cellular restriction mechanisms. We further show that viral replication occurs in a small but specific sub-group of cells transcriptionally related to, and likely derived from, a cluster of cells expressing markers of Colony Forming Unit - Granulocyte, Erythrocyte, Monocyte, Megakaryocyte (CFU-GEMM) oligopotent progenitors. Compared to the remainder of the population, CFU-GEMM cells are enriched in transcripts with functions in mitochondria' energy production, cell proliferation, RNA processing and protein synthesis, and express similar or higher levels of interferon-related genes. While expression levels of the former are maintained in infected cells, the latter are strongly down-regulated. We thus propose that the preferential infection of CFU-GEMM cells may be due to the presence of a preestablished pro-viral environment, requiring minimal optimization efforts from viral effectors, rather than to the absence of specific restriction factors. Together, these findings identify a potentially new population of myeloid cells permissive to CMV replication, and provide a possible rationale for their preferential infection.