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病毒去污方法对COVID-19患者细胞因子谱的影响

Impact of Viral Decontamination Method on Cytokine Profile of COVID-19 Patients.

作者信息

Magrì Davide, Navarro Anna, Bergami Federica, Percivalle Elena, Ferrari Alessandro, Lettieri Teresa, Calzolai Luigi, Piralla Antonio, Baldanti Fausto, Gioria Sabrina

机构信息

European Commission, Joint Research Centre (JRC), 21027 Ispra, Italy.

Molecular Virology Unit, Microbiology and Virology Department, Fondazione IRCCS Policlinico San Matteo, 27100 Pavia, Italy.

出版信息

Biomedicines. 2021 Sep 22;9(10):1287. doi: 10.3390/biomedicines9101287.

Abstract

COVID-19 related morbidity and mortality have been often attributed to an exaggerated immune response. The role of cytokines and chemokines in COVID-19 and their contributions to illness severity are known, and thus their profiling from patient bronchoalveolar lavage (BAL) samples would help in understanding the disease progression. To date, limited studies have been performed on COVID-19 BAL samples, as the manipulation of such specimens (potentially containing live viruses) requires several laboratorial precautions, such as personnel training and special equipment, a requirement that not all laboratories can fulfil. Here, we assessed two fast and easily applicable methods (ultrafiltration and ultraviolet-C irradiation) for their impact on viral load removal or inactivation, respectively and on cytokine profiles preservation. Eight samples of BAL fluids from SARS-CoV2 patients with high viral load were tested. For both methods, complete removal was confirmed by lack of viral replication in Vero E6 cells and by RT-qPCR. Although both methods showed to remove completely the active SARS-CoV2 viral load, only UVC treatment has little or no quantitative effect on total cytokines/chemokines measurements, however cytokines profile and relative ratios are preserved or minimally altered when compared data obtained by the two different decontamination methods. Sample preparation and manipulation can greatly affect the analytical results; therefore, understanding if changes occurred after sample processing is of outmost importance for reliable data and can be useful to improve clinical practice.

摘要

与新冠病毒相关的发病率和死亡率常常归因于过度的免疫反应。细胞因子和趋化因子在新冠病毒感染中的作用及其对疾病严重程度的影响已为人所知,因此对患者支气管肺泡灌洗(BAL)样本进行分析将有助于了解疾病进展。迄今为止,针对新冠病毒BAL样本的研究有限,因为处理此类样本(可能含有活病毒)需要采取多项实验室预防措施,如人员培训和特殊设备,而并非所有实验室都能满足这一要求。在此,我们评估了两种快速且易于应用的方法(超滤和紫外线-C照射),分别考察它们对病毒载量清除或灭活以及细胞因子谱保存的影响。对8份来自病毒载量高的新冠病毒患者的BAL液样本进行了检测。对于这两种方法,通过Vero E6细胞中无病毒复制以及逆转录定量聚合酶链反应(RT-qPCR)确认了病毒的完全清除。虽然两种方法均显示能完全清除活性新冠病毒载量,但只有紫外线-C处理对总细胞因子/趋化因子测量几乎没有定量影响,然而,当比较两种不同去污方法获得的数据时,细胞因子谱及其相对比例得以保留或仅有微小改变。样本制备和处理会极大地影响分析结果;因此,了解样本处理后是否发生变化对于获得可靠数据至关重要,并且有助于改善临床实践。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/74bc/8533265/83cd694c8375/biomedicines-09-01287-g001.jpg

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