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Journal of virological methods

Journal of virological methods

IF: 2.2
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Cell-based reporter assays for measurements of antibody-mediated cellular cytotoxicity and phagocytosis against SARS-CoV-2 spike protein

Published:1 September 2022 DOI: 10.1016/j.jviromet.2022.114564 PMID: 35671888
Yuting Hong , Huilin Guo , Min Wei , Yali Zhang , Mujin Fang , Tong Cheng , Zhiyong Li , Shengxiang Ge , Xiangyang Yao , Quan Yuan , Ningshao Xia

Abstract

The COVID-19 pandemic caused by SARS-CoV-2 infections has led to excess deaths worldwide. Neutralizing antibodies (nAbs) against viral spike protein acquired from natural infections or vaccinations contribute to protection against new- and re-infections. Besides neutralization, antibody-mediated cellular cytotoxicity (ADCC) and phagocytosis (ADCP) are also important for viral clearance. However, due to the lack of convenient methods, the ADCC and ADCP responses elicited by viral infections or vaccinations remain to be explored. Here, we developed cell-based assays using target cells stably expressing SARS-CoV-2 spikes and Jurkat-NFAT-CD16a/CD32a effector cells for ADCC/ADCP measurements of monoclonal antibodies and human convalescent COVID-19 plasmas (HCPs). In control samples (n?=?190), the specificity was 99.5% (95%CI: 98.4–100%) and 97.4% (95%CI: 95.1–99.6%) for the ADCC and ADCP assays, respectively. Among 87 COVID-19 HCPs, 83 (sensitivity: 95.4%, 95%CI: 91.0–99.8%) and 81 (sensitivity: 93.1%, 95%CI: 87.8–98.4%) showed detectable ADCC (titer range: 7.4–1721.6) and ADCP activities (titer range: 4–523.2). Notably, both ADCC and ADCP antibody titers positively correlated with the nAb titers in HCPs. In summary, we developed new tools for quantitative ADCC and ADCP analysis against SARS-CoV-2, which may facilitate further evaluations of Fc-mediated effector functions in preventing and treating against SARS-CoV-2.

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Materials
Procduct Name CAS Molecular Formula Supplier Price
Coelenterazine 55779-48-1 C26H21N3O3 252 suppliers $45.00-$14697.00
Coelenterazine 55779-48-1 C26H21N3O3 252 suppliers $45.00-$14697.00
Coelenterazine 55779-48-1 C26H21N3O3 252 suppliers $45.00-$14697.00
Coelenterazine 55779-48-1 C26H21N3O3 252 suppliers $45.00-$14697.00

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