A robust human norovirus replication model in zebrafish larvae
A robust human norovirus replication model in zebrafish larvae
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DOI：
10.1371/journal.ppat.1008009
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发表时间：
2019-09-01
期刊：
影响因子：
6.7
通讯作者：
Rocha-Pereira, Joana
中科院分区：
文献类型：
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作者：
Van Dycke, Jana;Ny, Annelii;Rocha-Pereira, Joana
人类诺如病毒（HuNoVs）是食源性疾病的最常见原因，每年造成600亿美元的社会成本和21.9万人死亡。缺乏强大的小动物模型严重阻碍了对诺如病毒生物学的理解和有效治疗方法的发展。在这里，我们报道了hunv GI和GII在斑马鱼（Danio rerio）幼虫中复制到高滴度；复制在感染后第2天达到高峰，至少6天内可检测到。该病毒（HuNoV GII.4）可在幼虫间连续传代2次。HuNoV在造血谱系和肠细胞中检测到，支持双向性的概念。抗病毒治疗使HuNoV的复制减少了bbbb2 log(10)，表明该模型适用于抗病毒研究。斑马鱼幼虫构成了一个简单而强大的复制模型，将在很大程度上促进HuNoV生物学的研究和抗病毒策略的开发。人类诺如病毒（HuNoV）是世界范围内病毒性胃肠炎的头号病原体。它可以感染所有年龄组的人，每年导致7亿人感染，21.9万人死亡。急性HuNoV胃肠炎的爆发经常发生，但慢性感染也发生在免疫缺陷的人群中。尽管具有临床意义，但由于没有简单有效的培养系统，研究该病毒一直是一项长达数十年的挑战。这种情况最近开始改变；我们在此通过建立一个体内模型系统来研究斑马鱼幼体的HuNoV复制，从而向前迈出了重要的一步。我们在3天龄的斑马鱼幼虫的蛋黄（食物储备）中注射HuNoV，结果在接种后6天病毒滴度很高。我们可以通过包括组织学在内的多种技术在受感染幼虫的组织中检测病毒，这些技术向我们展示了病毒存在于哪些器官。重要的是，通过在幼虫游泳的水中添加抗病毒药物，我们可以显著降低幼虫体内的病毒水平。这意味着从现在开始，测试小分子和开发首个针对HuNoV的抗病毒疗法将变得容易得多。
Human noroviruses (HuNoVs) are the most common cause of foodborne illness, with a societal cost of $60 billion and 219,000 deaths/year. The lack of robust small animal models has significantly hindered the understanding of norovirus biology and the development of effective therapeutics. Here we report that HuNoV GI and GII replicate to high titers in zebrafish (Danio rerio) larvae; replication peaks at day 2 post infection and is detectable for at least 6 days. The virus (HuNoV GII.4) could be passaged from larva to larva two consecutive times. HuNoV is detected in cells of the hematopoietic lineage and the intestine, supporting the notion of a dual tropism. Antiviral treatment reduces HuNoV replication by >2 log(10), showing that this model is suited for antiviral studies. Zebrafish larvae constitute a simple and robust replication model that will largely facilitate studies of HuNoV biology and the development of antiviral strategies.Author summary Human norovirus (HuNoV) is the number one agent of viral gastroenteritis worldwide. It can infect people of all age groups, resulting in 700 million infections and 219,000 deaths each year. Outbreaks of acute HuNoV gastroenteritis occur often, but chronic infections also happen in people with immune deficiencies. Despite its clinical relevance, studying the virus has been a decades-long challenge because no simple and efficient cultivation system existed. This has recently started to change; we here contribute with an important step forward by establishing an in vivo model system to study HuNoV replication using zebrafish larvae. We inject a HuNoV in the yolk (food reserve) of 3-day-old zebrafish larvae, which results in high virus titers up to 6 days after inoculation. We could detect the virus in the tissues of the infected larvae by a variety of techniques including histology, which showed us in which organs the virus is present. Importantly, by adding an antiviral to the water in which the larvae swim, we could significantly reduce the virus levels in the larvae. This means that testing small molecules and developing the first antiviral therapy for HuNoV will be much easier from hereon.