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新冠病毒患者鼻咽拭子中ACE2和TMPRSS2的免疫细胞化学评估

Immunocytochemical Assessment of ACE2 and TMPRSS2 in Nasopharyngeal Swabs from SARS-CoV-2 Patients.

作者信息

Peña Karla Beatríz, Gumà Josep, Guilarte Carmen, Delamo Lola, Grifol Marc, Pique Berta, Hernandez Anna, Casteñé Helena, Riu Francesc, Parada David

机构信息

Pathology Molecular Unit, Department of Pathology, Sant Joan University Hospital, Pere Virgili Health Research Institute, Faculty of Medicine, Rovira and Virgili University, 43204 Reus (Tarragona), Spain.

Department of Oncology, Sant Joan University Hospital, Pere Virgili Health Research Institute, Faculty of Medicine, Rovira and Virgili University, 43204 Reus (Tarragona), Spain.

出版信息

Front Biosci (Landmark Ed). 2022 Jul 11;27(7):217. doi: 10.31083/j.fbl2707217.

Abstract

BACKGROUND

SARS-CoV-2 is a positive-sense single-stranded RNA virus. It is enveloped by four structural proteins. The entry of the virus into the host cells is mediated by spike protein binding to the angiotensin converting enzyme 2 (ACE2) and proteolytic cleavage by transmembrane protease serine 2 (TMPRSS2). In this study, we analyzed the expression of the ACE2 receptor and TMPRSS2 in cases under investigation for SARS-CoV-2 infection.

METHODS

The study was carried out using the viral transport medium of consecutive nasopharyngeal swabs from 300 people under examination for SARS-CoV-2 infection. All samples underwent the SARS-CoV-2 transcriptase-mediated amplification assay (Procleix® SARS-CoV-2) to detect the virus. Immunocytochemistry was used in each sample to detect the presence of the SARS-CoV-2 nucleoprotein, the ACE2 receptor, and TMPRSS2.

RESULTS

An immunocytochemical study with monoclonal antibody against SARS-CoV-2 viral nucleoprotein showed positivity in squamous cells. ACE2 were not detected in the squamous cells obtained from the nasopharyngeal samples.

CONCLUSIONS

SARS-CoV-2 predominantly localizes to squamous cells in cytology samples of patients with positive transcriptase-mediated amplification SARS-CoV-2 assay results. The immunocytochemical negativity for ACE2 evidenced in the present study could be related to the cellular heterogeneity present in the nasopharyngeal smear samples and could be related to variations at the genomic level. Our results suggest that SARS-CoV-2 might be present in the nasopharyngeal region because viral cell junctions are weaker. This facilitates viral concentration, infective capacity and migration to specific organs, where SARS-CoV-2 infects target cells by binding to their receptors and then entering.

摘要

背景

严重急性呼吸综合征冠状病毒2(SARS-CoV-2)是一种正链单股RNA病毒。它由四种结构蛋白包裹。病毒进入宿主细胞是通过刺突蛋白与血管紧张素转换酶2(ACE2)结合,并由跨膜蛋白酶丝氨酸2(TMPRSS2)进行蛋白水解切割来介导的。在本研究中,我们分析了疑似SARS-CoV-2感染病例中ACE2受体和TMPRSS2的表达情况。

方法

本研究使用了来自300名接受SARS-CoV-2感染检测者连续鼻咽拭子的病毒运输培养基。所有样本均进行了SARS-CoV-2转录介导扩增检测(Procleix® SARS-CoV-2)以检测病毒。对每个样本进行免疫细胞化学检测,以检测SARS-CoV-2核蛋白、ACE2受体和TMPRSS2的存在情况。

结果

用抗SARS-CoV-2病毒核蛋白的单克隆抗体进行的免疫细胞化学研究显示鳞状细胞呈阳性。从鼻咽样本获得的鳞状细胞中未检测到ACE2。

结论

在转录介导扩增SARS-CoV-2检测结果为阳性的患者细胞学样本中,SARS-CoV-2主要定位于鳞状细胞。本研究中ACE2的免疫细胞化学阴性可能与鼻咽涂片样本中存在的细胞异质性有关,也可能与基因组水平的变异有关。我们的结果表明,SARS-CoV-2可能存在于鼻咽区域,因为病毒细胞连接较弱。这有利于病毒聚集、感染能力以及向特定器官迁移,在这些器官中SARS-CoV-2通过与其受体结合然后进入来感染靶细胞。

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