Institute of Human Virology and Global Virus Network Center, Department of Biochemistry and Molecular Biology, School of Medicine, University of Maryland, Baltimore, MD 21201, USA.
Institute of Human Virology and Global Virus Network Center, Department of Medicine, School of Medicine, University of Maryland, Baltimore, MD 21201, USA.
Viruses. 2021 Oct 30;13(11):2193. doi: 10.3390/v13112193.
SARS-CoV-2 (Severe Acute Respiratory Syndrome Coronavirus 2) is primarily responsible for coronavirus disease (COVID-19) and it is characterized by respiratory illness with fever and dyspnea. Severe vascular problems and several other manifestations, including neurological ones, have also been frequently reported, particularly in the great majority of "long hauler" patients. SARS-CoV-2 infects and replicates in lung epithelial cells, while dysfunction of endothelial and neuronal brain cells has been observed in the absence of productive infection. It has been shown that the Spike protein can interact with specific cellular receptors, supporting both viral entry and cellular dysfunction. It is thus clear that understanding how and when these receptors are regulated, as well as how much they are expressed would help in unveiling the multifaceted aspects of this disease. Here, we show that SH-SY5Y neuroblastoma cells express three important cellular surface molecules that interact with the Spike protein, namely ACE2, TMPRSS2, and NRP1. Their levels increase when cells are treated with retinoic acid (RA), a commonly used agent known to promote differentiation. This increase matched the higher levels of receptors observed on HUVEC (primary human umbilical vein endothelial cells). We also show by confocal imaging that replication-defective pseudoviruses carrying the SARS-CoV-2 Spike protein can infect differentiated and undifferentiated SH-SY5Y, and HUVEC cells, although with different efficiencies. Neuronal cells and endothelial cells are potential targets for SARS-CoV-2 infection and the interaction of the Spike viral protein with these cells may cause their dysregulation. Characterizing RNA and protein expression tempo, mode, and levels of different SARS-CoV-2 receptors on both cell subpopulations may have clinical relevance for the diagnosis and treatment of COVID-19-infected subjects, including long hauler patients with neurological manifestations.
严重急性呼吸综合征冠状病毒 2（SARS-CoV-2）是导致冠状病毒病（COVID-19）的主要病原体，其特征是发热和呼吸困难的呼吸道疾病。严重的血管问题和其他几种表现，包括神经系统表现，也经常被报道，特别是在绝大多数“长程患者”中。SARS-CoV-2 感染和在肺上皮细胞中复制，而在没有有效感染的情况下观察到内皮细胞和神经元脑细胞的功能障碍。已经表明，刺突蛋白可以与特定的细胞受体相互作用，支持病毒进入和细胞功能障碍。因此，很明显，了解这些受体是如何以及何时受到调节，以及它们的表达量如何，将有助于揭示这种疾病的多方面特征。在这里，我们表明，SH-SY5Y 神经母细胞瘤细胞表达三种与刺突蛋白相互作用的重要细胞表面分子，即 ACE2、TMPRSS2 和 NRP1。当细胞用维甲酸（RA）处理时，它们的水平会增加，RA 是一种常用的促进分化的试剂。这种增加与在 HUVEC（原代人脐静脉内皮细胞）上观察到的更高水平的受体相匹配。我们还通过共聚焦成像显示，携带 SARS-CoV-2 刺突蛋白的复制缺陷型假病毒可以感染分化和未分化的 SH-SY5Y 和 HUVEC 细胞，尽管效率不同。神经元细胞和内皮细胞是 SARS-CoV-2 感染的潜在靶细胞，刺突病毒蛋白与这些细胞的相互作用可能导致它们的失调。表征两种细胞亚群上不同 SARS-CoV-2 受体的 RNA 和蛋白表达的时间、模式和水平可能对 COVID-19 感染患者的诊断和治疗具有临床意义，包括具有神经系统表现的长程患者。
