Activity of human serum antibodies in an influenza virus hemagglutinin stalk-based ADCC reporter assay correlates with activity in a CD107a degranulation assay
Activity of human serum antibodies in an influenza virus hemagglutinin stalk-based ADCC reporter assay correlates with activity in a CD107a degranulation assay
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DOI：
10.1016/j.vaccine.2020.01.008
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发表时间：
2020-02-18
期刊：
影响因子：
5.5
通讯作者：
Krammer, Florian
中科院分区：
文献类型：
--
作者：
Chromikova, Veronika;Tan, Jessica;Krammer, Florian
流感病毒血凝素（HA）的茎是基于抗体的通用流感病毒疫苗开发的一个有吸引力的靶点。虽然针对病毒这部分的抗体可以被中和，但近年来研究表明，Fc受体介导的效应物功能对抗茎抗体的保护作用具有重要意义。目前存在几种测量Fc-Fc受体相互作用效应功能（如抗体依赖性细胞毒性和抗体依赖性细胞吞噬）的方法，但它们存在低通量和高运行间可变性等局限性。基于报告细胞表达荧光素酶的人Fc γ RIIIa与抗原结合抗体Fc结合的报告细胞，也开发了抗体依赖性细胞毒性的报告细胞测定方法。这些报告基因检测方法可以在高通量环境中使用，但由于它们只表达一种Fc受体，因此其生物学相关性尚不清楚。在这里，我们优化了抗体依赖的细胞毒性报告试验，以测量H1血凝素保守柄结构域抗体的活性。然后将该检测与基于cd107的脱颗粒检测相关联，并且可以观察到强烈且显著的相关性。这些数据表明，基于Fc γ riiia的报告基因检测是功能检测的良好替代品，特别是在需要分析大量样本量的情况下。(C) 2020 Elsevier Ltd.版权所有。
The stalk of the influenza virus hemagglutinin (HA) is an attractive target for antibody-based universal influenza virus vaccine development. While antibodies that target this part of the virus can be neutralizing, it has been shown in recent years that Fc receptor-mediated effector functions are of significant importance for the protective effect of anti-stalk antibodies. Several assays to measure Fc-Fc receptor interaction-based effector functions like antibody-dependent cellular cytotoxicity and antibody-dependent cellular phagocytosis exist, but they suffer from limitations such as low throughput and high run-to-run variability. Reporter assays for antibody-dependent cellular cytotoxicity based on reporter cells that express luciferase upon engagement of human Fc gamma RIIIa with the Fc of antigen-bound antibodies have been developed as well. These reporter assays can be used in a higher throughput setting with limited run-to-run assay variability but since they express only one Fc receptor, their biological relevance is unclear. Here we optimized an antibody-dependent cellular cytotoxicity reporter assay to measure the activity of antibodies to the conserved stalk domain of H1 hemagglutinin. The assay was then correlated to a CD107a-based degranulation assay, and a strong and significant correlation could be observed. This data suggests that the Fc gamma RIIIa-based reporter assay is a good substitute for functional assays, especially in settings where larger sample numbers need to be analyzed. (C) 2020 Elsevier Ltd. All rights reserved.