Rowntree Louise C, Chua Brendon Y, Nicholson Suellen, Koutsakos Marios, Hensen Luca, Douros Celia, Selva Kevin, Mordant Francesca L, Wong Chinn Yi, Habel Jennifer R, Zhang Wuji, Jia Xiaoxiao, Allen Lily, Doolan Denise L, Jackson David C, Wheatley Adam K, Kent Stephen J, Amanat Fatima, Krammer Florian, Subbarao Kanta, Cheng Allen C, Chung Amy W, Catton Mike, Nguyen Thi Ho, van de Sandt Carolien E, Kedzierska Katherine
Department of Microbiology and Immunology University of Melbourne, at the Peter Doherty Institute for Infection and Immunity Melbourne VIC Australia.
Global Station for Zoonosis Control Global Institution for Collaborative Research and Education (GI-CoRE) Hokkaido University Sapporo Hokkaido Japan.
Clin Transl Immunology. 2021 Feb 28;10(3):e1258. doi: 10.1002/cti2.1258. eCollection 2021.
As the world transitions into a new era of the COVID-19 pandemic in which vaccines become available, there is an increasing demand for rapid reliable serological testing to identify individuals with levels of immunity considered protective by infection or vaccination.
We used 34 SARS-CoV-2 samples to perform a rapid surrogate virus neutralisation test (sVNT), applicable to many laboratories as it circumvents the need for biosafety level-3 containment. We correlated results from the sVNT with five additional commonly used SARS-CoV-2 serology techniques: the microneutralisation test (MNT), in-house ELISAs, commercial Euroimmun- and Wantai-based ELISAs (RBD, spike and nucleoprotein; IgG, IgA and IgM), antigen-binding avidity, and high-throughput multiplex analyses to profile isotype, subclass and Fc effector binding potential. We correlated antibody levels with antibody-secreting cell (ASC) and circulatory T follicular helper (cTfh) cell numbers.
Antibody data obtained with commercial ELISAs closely reflected results using in-house ELISAs against RBD and spike. A correlation matrix across ten measured ELISA parameters revealed positive correlations for all factors. The frequency of inhibition by rapid sVNT strongly correlated with spike-specific IgG and IgA titres detected by both commercial and in-house ELISAs, and MNT titres. Multiplex analyses revealed strongest correlations between IgG, IgG1, FcR and C1q specific to spike and RBD. Acute cTfh-type 1 cell numbers correlated with spike and RBD-specific IgG antibodies measured by ELISAs and sVNT.
Our comprehensive analyses provide important insights into SARS-CoV-2 humoral immunity across distinct serology assays and their applicability for specific research and/or diagnostic questions to assess SARS-CoV-2-specific humoral responses.
随着世界步入新冠疫情的新时代，疫苗已可获取，对快速可靠的血清学检测的需求日益增加，以识别具有经感染或疫苗接种产生的保护性免疫水平的个体。
我们使用34份严重急性呼吸综合征冠状病毒2（SARS-CoV-2）样本进行了快速替代病毒中和试验（sVNT），该试验适用于许多实验室，因为它无需生物安全3级防护。我们将sVNT的结果与另外五种常用的SARS-CoV-2血清学技术的结果进行了关联：微量中和试验（MNT）、内部酶联免疫吸附测定（ELISA）、基于欧蒙和万泰的商业ELISA（针对受体结合域（RBD）、刺突蛋白和核蛋白；检测IgG、IgA和IgM）、抗原结合亲和力以及高通量多重分析以分析同种型、亚类和Fc效应器结合潜力。我们将抗体水平与抗体分泌细胞（ASC）和循环滤泡辅助性T细胞（cTfh）数量进行了关联。
商业ELISA获得的抗体数据与使用针对RBD和刺突蛋白的内部ELISA的结果密切相关。在十个测量的ELISA参数的相关矩阵中，所有因素均呈正相关。快速sVNT的抑制频率与商业和内部ELISA检测到的刺突蛋白特异性IgG和IgA滴度以及MNT滴度密切相关。多重分析显示，刺突蛋白和RBD特异性的IgG、IgG1、FcR和C1q之间的相关性最强。急性cTfh 1型细胞数量与ELISA和sVNT测量的刺突蛋白和RBD特异性IgG抗体相关。
我们的综合分析为不同血清学检测方法中的SARS-CoV-2体液免疫及其在评估SARS-CoV-2特异性体液反应的特定研究和/或诊断问题中的适用性提供了重要见解。
