One for all-human kidney Caki-1 cells are highly susceptible to infection with corona- and other respiratory viruses
One for all-human kidney Caki-1 cells are highly susceptible to infection with corona- and other respiratory viruses
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DOI：
10.1128/jvi.00555-23
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发表时间：
2023-09-28
影响因子：
5.4
通讯作者：
Tait-Burkard，Christine
中科院分区：
文献类型：
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作者：
Daniels，Alison;Fletcher，Sarah;Tait-Burkard，Christine
在体外研究宿主-病毒相互作用依赖于合适的细胞和组织培养模型。结果只与生成它们的模型一样好。然而，选择用于体外工作的细胞模型通常仅取决于可用性和先前的使用。尽管过去几年冠状病毒研究大幅增加，但科学家们仍然严重依赖：非人类、高度异质或未完全分化的细胞，或需要过表达受体和其他辅助因子的天然不敏感细胞。复杂的原代或干细胞模型是高度代表性的人体组织，但昂贵和耗时的开发和维护有限的适用性高通量experiments.Using组织特异性表达模式，我们确定了人肾细胞作为一个理想的目标严重急性呼吸综合征冠状病毒-2（SARS-CoV-2）和更广泛的冠状病毒感染。我们展示了使用特征良好的人肾细胞系Caki-1感染三种人冠状病毒（hCoV）：β冠状病毒SARS-CoV-2、中东呼吸综合征冠状病毒和α冠状病毒CoV 229 E。当与用于培养相应病毒的其他常用细胞系相比时，Caki-1细胞显示出对所有三种冠状病毒的相等或上级易感性。针对SARS-CoV-2 N蛋白的抗体染色显示出相当的复制率。一组26种定制抗体显示了SARS-CoV-2蛋白在复制过程中使用免疫细胞化学的位置。此外，Caki-1细胞还被发现对其他两种人类呼吸道病毒（A型流感病毒和呼吸道合胞病毒）敏感，这使其成为广泛的呼吸道病毒交叉比较的理想模型。重要的是，细胞系仍然是病毒研究的支柱，但结果仅与其原始模型一样好。尽管在COVID-19大流行之后对人类冠状病毒的研究有所增加，但研究人员继续依赖于非人类起源、不完全分化或缺乏活性干扰素反应的次优细胞系模型。我们确定了人肾Caki-1细胞系作为严重急性呼吸综合征冠状病毒-2（SARS-CoV-2）的潜在靶点。该细胞系可被广泛的冠状病毒感染，包括普通感冒病毒hCoV-229 E、流行性病毒MERS-CoV和SARS-CoV-2以及其他重要的呼吸道病毒甲型流感病毒和呼吸道合胞病毒。我们可以显示26个SARS-CoV-2蛋白在Caki-1细胞中的定位在自然复制过程中，并且细胞能够形成细胞免疫应答。总之，这使得Caki-1细胞成为在一个细胞系中进行跨病毒比较的独特工具。
In vitroinvestigations of host-virus interactions are reliant on suitable cell and tissue culture models. Results are only as good as the model they are generated in. However, choosing cell models forin vitrowork often depends on availability and previous use alone. Despite the vast increase in coronavirus research over the past few years, scientists are still heavily reliant on: non-human, highly heterogeneous or not fully differentiated, or naturally unsusceptible cells requiring overexpression of receptors and other accessory factors. Complex primary or stem cell models are highly representative of human tissues but are expensive and time-consuming to develop and maintain with limited suitability for high-throughput experiments.Using tissue-specific expression patterns, we identified human kidney cells as an ideal target for severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) and broader coronavirus infection. We show the use of the well-characterized human kidney cell line Caki-1 for infection with three human coronaviruses (hCoVs):BetacoronavirusesSARS-CoV-2 and Middle Eastern respiratory syndrome coronavirus andAlphacoronavirushCoV 229E. Caki-1 cells show equal or superior susceptibility to all three coronaviruses when compared to other commonly used cell lines for the cultivation of the respective virus. Antibody staining against SARS-CoV-2 N protein shows comparable replication rates. A panel of 26 custom antibodies shows the location of SARS-CoV-2 proteins during replication using immunocytochemistry. In addition, Caki-1 cells were found to be susceptible to two other human respiratory viruses, influenza A virus and respiratory syncytial virus, making them an ideal model for cross-comparison for a broad range of respiratory viruses.IMPORTANCECell lines remain the backbone of virus research, but results are only as good as their originating model. Despite increased research into human coronaviruses following the COVID-19 pandemic, researchers continue to rely on suboptimal cell line models of: non-human origin, incomplete differentiation, or lacking active interferon responses. We identified the human kidney Caki-1 cell line as a potential target for severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2). This cell line could be shown to be infectable with a wide range of coronaviruses including common cold virus hCoV-229E, epidemic virus MERS-CoV, and SARS-CoV-2 as well as other important respiratory viruses influenza A virus and respiratory syncytial virus. We could show the localization of 26 SARS-CoV-2 proteins in Caki-1 cells during natural replication and the cells are competent of forming a cellular immune response. Together, this makes Caki-1 cells a unique tool for cross-virus comparison in one cell line.