K18-hACE2 mice develop respiratory disease resembling severe COVID-19.
K18-hACE2 mice develop respiratory disease resembling severe COVID-19.
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DOI：
10.1371/journal.ppat.1009195
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发表时间：
2021-01
期刊：
影响因子：
6.7
通讯作者：
Munster VJ
中科院分区：
文献类型：
--
作者：
Yinda CK;Port JR;Bushmaker T;Offei Owusu I;Purushotham JN;Avanzato VA;Fischer RJ;Schulz JE;Holbrook MG;Hebner MJ;Rosenke R;Thomas T;Marzi A;Best SM;de Wit E;Shaia C;van Doremalen N;Munster VJ
SARS-CoV-2于2019年底出现，并导致了持续的COVID-19大流行。一些动物模型已经迅速发展，概括无症状到中度疾病谱系。目前，迫切需要更多的小动物模型来研究COVID-19重症发病机制和快速制定医疗对策。在这里，我们发现在细胞角蛋白18启动子（K18）下表达人SARS-CoV-2受体（血管紧张素转换酶2 [hACE2]）的转基因小鼠对SARS-CoV-2易感，并且感染导致剂量依赖的致死过程。接种104 TCID50或105 TCID50后，两组小鼠体重均迅速下降，105 TCID50组小鼠死亡率一致。在上呼吸道和下呼吸道观察到高水平的病毒RNA脱落，在肠道观察到间歇性脱落。接种SARS-CoV-2导致上呼吸道和下呼吸道感染，鼻甲、气管和肺部感染病毒滴度高。观察到的间质性肺炎和肺部病理与SARS-CoV-2在肺细胞中明显复制相似，与COVID-19重症病例相似。SARS-CoV-2感染导致肺部巨噬细胞和淋巴细胞浸润，Th1和促炎细胞因子/趋化因子上调。在7 DPI时，在一些动物的大脑皮层和海马中观察到SARS-CoV-2的肺外复制，而在3 DPI时则没有。快速的炎症反应和观察到的病理与COVID-19相似。此外，我们证明低剂量感染102 TCID50 SARS-CoV-2可以模拟轻度病程，导致最小的临床表现和接近均匀的生存。综上所述，这些数据支持了该模型在未来发病机制研究和医学对策开发中的应用。COVID-19在人类中的疾病表现从无症状到严重不等。虽然已经开发了几种轻度至中度疾病模型，但仍然需要能够概括在一部分患者中观察到的严重和致命进展的动物模型。本研究表明，人源化转基因小鼠在接种了COVID-19的病原SARS-CoV-2后出现了剂量依赖性疾病。接种后第3天，小鼠出现上呼吸道和下呼吸道感染，病毒也在脑内复制。在这些小鼠中观察到的病理和免疫疾病表现与人类COVID-19相似。这表明该模型在阐明COVID-19发病机制和测试对策方面越来越有用，这两者都是迫切需要的。
SARS-CoV-2 emerged in late 2019 and resulted in the ongoing COVID-19 pandemic. Several animal models have been rapidly developed that recapitulate the asymptomatic to moderate disease spectrum. Now, there is a direct need for additional small animal models to study the pathogenesis of severe COVID-19 and for fast-tracked medical countermeasure development. Here, we show that transgenic mice expressing the human SARS-CoV-2 receptor (angiotensin-converting enzyme 2 [hACE2]) under a cytokeratin 18 promoter (K18) are susceptible to SARS-CoV-2 and that infection resulted in a dose-dependent lethal disease course. After inoculation with either 104 TCID50 or 105 TCID50, the SARS-CoV-2 infection resulted in rapid weight loss in both groups and uniform lethality in the 105 TCID50 group. High levels of viral RNA shedding were observed from the upper and lower respiratory tract and intermittent shedding was observed from the intestinal tract. Inoculation with SARS-CoV-2 resulted in upper and lower respiratory tract infection with high infectious virus titers in nasal turbinates, trachea and lungs. The observed interstitial pneumonia and pulmonary pathology, with SARS-CoV-2 replication evident in pneumocytes, were similar to that reported in severe cases of COVID-19. SARS-CoV-2 infection resulted in macrophage and lymphocyte infiltration in the lungs and upregulation of Th1 and proinflammatory cytokines/chemokines. Extrapulmonary replication of SARS-CoV-2 was observed in the cerebral cortex and hippocampus of several animals at 7 DPI but not at 3 DPI. The rapid inflammatory response and observed pathology bears resemblance to COVID-19. Additionally, we demonstrate that a mild disease course can be simulated by low dose infection with 102 TCID50 SARS-CoV-2, resulting in minimal clinical manifestation and near uniform survival. Taken together, these data support future application of this model to studies of pathogenesis and medical countermeasure development. The disease manifestation of COVID-19 in humans ranges from asymptomatic to severe. While several mild to moderate disease models have been developed, there is still a need for animal models that recapitulate the severe and fatal progression observed in a subset of patients. Here, we show that humanized transgenic mice developed dose-dependent disease when inoculated with SARS-CoV-2, the etiological agent of COVID-19. The mice developed upper and lower respiratory tract infection, with virus replication also in the brain after day 3 post inoculation. The pathological and immunological diseases manifestation observed in these mice bears resemblance to human COVID-19. This suggests increased usefulness of this model for elucidating COVID-19 pathogenesis and for testing of countermeasures, both of which are urgently needed.
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