A Bifluorescent-Based Assay for the Identification of Neutralizing Antibodies against SARS-CoV-2 Variants of Concern In Vitro and In Vivo.
A Bifluorescent-Based Assay for the Identification of Neutralizing Antibodies against SARS-CoV-2 Variants of Concern In Vitro and In Vivo.
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DOI：
10.1128/jvi.01126-21
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发表时间：
2021-10-27
影响因子：
5.4
通讯作者：
Martinez-Sobrido L
中科院分区：
文献类型：
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作者：
Chiem K;Morales Vasquez D;Silvas JA;Park JG;Piepenbrink MS;Sourimant J;Lin MJ;Greninger AL;Plemper RK;Torrelles JB;Walter MR;de la Torre JC;Kobie JK;Ye C;Martinez-Sobrido L
严重急性呼吸综合征冠状病毒2型（SARS-CoV-2）于2019年底出现，是目前仍在持续的2019冠状病毒病（COVID-19）大流行的罪魁祸首。美国食品和药物管理局（FDA）已批准预防COVID-19的预防性疫苗。鉴定SARS-CoV-2中和抗体（nab）对于评估疫苗的保护效果非常重要，包括它们对新出现的SARS-CoV-2关注变体（VoC）的保护能力。在这里，我们报道了一种重组(r)SARS-CoV-2 USA/WA1/2020 （WA-1）菌株表达Venus和一种rSARS-CoV-2菌株表达mCherry，并在南非（SA） B.1.351 (β [β]) VoC的刺突(S)糖蛋白的受体结合域（RBD）中发现了K417N、E484K和N501Y突变，通过双荧光法快速准确地鉴定出能够在体外和体内中和病毒感染的人单克隆抗体（hmab）。重要的是，我们基于双荧光的系统准确地概括了使用单个病毒观察到的结果。此外，荧光表达的SARS-CoV-2菌株与亲本野生型（WT） SARS-CoV-2 WA-1菌株在体外具有相似的病毒适应度，在K18人血管紧张素转换酶2 （hACE2）转基因小鼠感染SARS-CoV-2模型中具有相似的体内毒力和致病性。我们证明，这些新的荧光表达的SARS-CoV-2可以在体外和体内容易地鉴定同时中和不同SARS-CoV-2菌株（包括VoC）的hmab，用于快速评估疫苗效力或鉴定用于治疗SARS-CoV-2感染的预防性和/或治疗性广泛nab。SARS-CoV-2是导致日常生活和社会经济扭曲的COVID-19大流行的罪魁祸首。目前仍迫切需要预防和治疗SARS-CoV-2感染的药物。在这项研究中，我们证明了使用基于双荧光的检测方法快速鉴定对SARS-CoV-2具有中和活性的hmab的可行性，包括体外和体内的VoC。重要的是，通过这些基于双荧光的检测获得的结果概括了在单个病毒中观察到的结果，证明了它们的可行性，可以快速提高我们对疫苗功效的理解，并确定用于治疗SARS-CoV-2的广泛保护性人类抗体。
Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) emerged at the end of 2019 and has been responsible for the still ongoing coronavirus disease 2019 (COVID-19) pandemic. Prophylactic vaccines have been authorized by the U.S. Food and Drug Administration (FDA) for the prevention of COVID-19. Identification of SARS-CoV-2-neutralizing antibodies (NAbs) is important to assess vaccine protection efficacy, including their ability to protect against emerging SARS-CoV-2 variants of concern (VoC). Here, we report the generation and use of a recombinant (r)SARS-CoV-2 USA/WA1/2020 (WA-1) strain expressing Venus and an rSARS-CoV-2 strain expressing mCherry and containing mutations K417N, E484K, and N501Y found in the receptor binding domain (RBD) of the spike (S) glycoprotein of the South African (SA) B.1.351 (beta [β]) VoC in bifluorescent-based assays to rapidly and accurately identify human monoclonal antibodies (hMAbs) able to neutralize both viral infections in vitro and in vivo. Importantly, our bifluorescent-based system accurately recapitulated findings observed using individual viruses. Moreover, fluorescent-expressing rSARS-CoV-2 strain and the parental wild-type (WT) rSARS-CoV-2 WA-1 strain had similar viral fitness in vitro, as well as similar virulence and pathogenicity in vivo in the K18 human angiotensin-converting enzyme 2 (hACE2) transgenic mouse model of SARS-CoV-2 infection. We demonstrate that these new fluorescent-expressing rSARS-CoV-2 can be used in vitro and in vivo to easily identify hMAbs that simultaneously neutralize different SARS-CoV-2 strains, including VoC, for the rapid assessment of vaccine efficacy or the identification of prophylactic and/or therapeutic broadly NAbs for the treatment of SARS-CoV-2 infection. IMPORTANCE SARS-CoV-2 is responsible of the COVID-19 pandemic that has warped daily routines and socioeconomics. There is still an urgent need for prophylactics and therapeutics to treat SARS-CoV-2 infections. In this study, we demonstrate the feasibility of using bifluorescent-based assays for the rapid identification of hMAbs with neutralizing activity against SARS-CoV-2, including VoC in vitro and in vivo. Importantly, results obtained with these bifluorescent-based assays recapitulate those observed with individual viruses, demonstrating their feasibility to rapidly advance our understanding of vaccine efficacy and to identify broadly protective human NAbs for the therapeutic treatment of SARS-CoV-2.