PDF(Pubmed)
Abstract：
The recent development of single cell sequencing technologies has revolutionized the state-of-art of cell biology, allowing the simultaneous measurement of thousands of genes in single cells. This technology has been applied to study the transcriptome of single cells in homeostasis and also in response to pathogenic exposure, greatly increasing our knowledge of the immune response to infectious agents. Yet the number of these studies performed in aquacultured fish species is still very limited. Thus, in the current study, we have used the 10x Genomics single cell RNA sequencing technology to study the response of rainbow trout (Oncorhynchus mykiss) peripheral blood leukocytes (PBLs) to infectious pancreatic necrosis virus (IPNV), an important trout pathogen. The study allowed us to obtain a transcriptomic profile of 12 transcriptionally distinct leukocyte cell subpopulations that included four different subsets of B cells, T cells, monocytes, two populations of dendritic-like cells (DCs), hematopoietic progenitor cells, non-specific cytotoxic cells (NCC), neutrophils and thrombocytes. The transcriptional pattern of these leukocyte subpopulations was compared in PBL cultures that had been exposed in vitro to IPNV for 24 h and mock-infected cultures. Our results revealed that monocytes and neutrophils showed the highest number of upregulated protein-coding genes in response to IPNV. Interestingly, IgM+IgD+ and IgT+ B cells also upregulated an important number of genes to the virus, but a much fainter response was observed in ccl4 + or plasma-like cells (irf4 + cells). A substantial number of protein-coding genes and genes coding for ribosomal proteins were also transcriptionally upregulated in response to IPNV in T cells and thrombocytes. Interestingly, although genes coding for ribosomal proteins were regulated in all affected PBL subpopulations, the number of such genes transcriptionally regulated was higher in IgM+IgD+ and IgT+ B cells. A further analysis dissected which of the regulated genes were common and which were specific to the different cell clusters, identifying eight genes that were transcriptionally upregulated in all the affected groups. The data provided constitutes a comprehensive transcriptional perspective of how the different leukocyte populations present in blood respond to an early viral encounter in fish.
摘要：
单细胞测序技术的最新发展彻底改变了细胞生物学的最先进水平，允许同时测量单个细胞中的数千个基因。该技术已用于研究稳态中以及对病原体暴露的反应中单细胞的转录组，大大增加了我们对感染因子的免疫反应的认识。然而，在水产养殖鱼类中进行的这些研究的数量仍然非常有限。因此，在目前的研究中，我们使用10x基因组学单细胞RNA测序技术研究了虹鳟鱼（Oncorhynchusmykiss）外周血白细胞（PBLs）对传染性胰腺坏死病毒（IPNV）的反应，一种重要的鳟鱼病原体。该研究使我们获得了12个转录上不同的白细胞亚群的转录组学图谱，其中包括四个不同的B细胞亚群，T细胞,单核细胞,两个树突状细胞（DC），造血祖细胞，非特异性细胞毒性细胞（NCC），中性粒细胞和血小板。在体外暴露于IPNV24小时的PBL培养物中和模拟感染的培养物中比较了这些白细胞亚群的转录模式。我们的结果表明，单核细胞和嗜中性粒细胞在响应IPNV时显示出最高数量的上调的蛋白质编码基因。有趣的是,IgM+IgD+和IgT+B细胞也上调了病毒重要数目的基因，但是在ccl4+或血浆样细胞（irf4+细胞）中观察到更微弱的反应。大量的蛋白质编码基因和核糖体蛋白编码基因也在T细胞和血小板中响应IPNV而被转录上调。有趣的是,尽管编码核糖体蛋白的基因在所有受影响的PBL亚群中都受到调控，在IgM+IgD+和IgT+B细胞中转录调节的这类基因数量较高。进一步的分析解剖了哪些受调控的基因是共同的，哪些是特定于不同的细胞簇，鉴定在所有受影响的组中转录上调的八个基因。所提供的数据构成了血液中存在的不同白细胞群体如何响应鱼类早期病毒遭遇的全面转录观点。
