Rapid and Flexible Platform To Assess Anti-SARS-CoV-2 Antibody Neutralization and Spike Protein-Specific Antivirals.
Rapid and Flexible Platform To Assess Anti-SARS-CoV-2 Antibody Neutralization and Spike Protein-Specific Antivirals.
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DOI：
10.1128/msphere.00571-21
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发表时间：
2021-08-25
期刊：
影响因子：
4.8
通讯作者：
Porotto M
中科院分区：
文献类型：
--
作者：
Stelitano D;Weisberg SP;Goldklang MP;Zhu Y;Bovier FT;Kalantarov GF;Greco G;Decimo D;Franci G;Cennamo M;Portella G;Galdiero M;Mathieu C;Horvat B;Trakht IN;Moscona A;Whitt MA;Porotto M
由严重急性呼吸综合征冠状病毒2型（SARS-CoV-2）引起的COVID-19大流行正在进行中，并向社会表明，需要灵活的方法来快速识别和筛选候选抗病毒药物。评估人血清的病毒中和活性以监测人群免疫和对感染和疫苗接种的反应是大流行控制的关键。我们开发了一个病毒中和平台策略，仅依赖于感兴趣的病毒的生物信息学和遗传信息。该平台使用病毒包膜糖蛋白cdna来建立一种模拟多周期感染的测定方法，但它是安全的，因此，对于需要BSL3设施的病毒（例如SARS-CoV-1和SARS-CoV-2），它符合生物安全2级（BSL2）条件。作为该平台的补充，我们提出了一种新的基于细胞的免疫荧光（CBI）检测方法，该方法使用表达SARS-CoV-2刺突蛋白(S)的细胞来准确测量人血清的中和潜力，并且易于适应相关变体。这些方法应该是评估抗病毒免疫（无论是通过自然感染还是通过疫苗获得）工具的有用补充。迫切需要通过自然感染或通过疫苗接种获得的抗体的中和特性的快速生物安全2级（BSL2）评价方法。在这里，我们提出了一种组合方法，使用基于细胞的免疫荧光（CBI）试验筛选血清中SARS-CoV-2刺突蛋白(S)的结合，并在BSL2条件下的伪病毒多周期感染模拟方案中进一步评估阳性样本。
The COVID-19 pandemic caused by severe acute respiratory syndrome coronavirus type 2 (SARS-CoV-2) is ongoing and has shown the community that flexible methods for rapidly identifying and screening candidate antivirals are needed. Assessing virus-neutralizing activity of human serum to monitor population immunity and response to infection and vaccination is key to pandemic control. We developed a virus neutralization platform strategy that relies only on bioinformatic and genetic information of the virus of interest. The platform uses viral envelope glycoprotein cDNAs to set up an assay that mimics multicycle infection but is safe and, therefore, amenable to biosafety level 2 (BSL2) conditions for viruses that require BSL3 facilities (e.g., SARS-CoV-1 and SARS-CoV-2). As a complement to this platform, we present a new cell-based immunofluorescent (CBI) assay that uses SARS-CoV-2 spike protein (S)-expressing cells to accurately measure the neutralization potential of human sera and is readily adaptable to variants of concern. These methods should be useful additions to the tools for assessing antiviral immunity, whether acquired via natural infection or vaccines. IMPORTANCE Assays for rapid biosafety level 2 (BSL2) evaluation of neutralizing properties of antibodies acquired via natural infection or through vaccination is urgently needed. Here, we propose a combinatorial approach in which sera are screened for SARS-CoV-2 spike protein (S) binding using a cell-based immunofluorescent (CBI) assay, and positive samples are further evaluated in a pseudotyped viral multicycle infection-mimicking protocol under BSL2 conditions.