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一种新的流式细胞术检测方法,用于测量针对表达 SARS-CoV-2 刺突蛋白的细胞的抗体依赖性细胞细胞毒性。

A new flow cytometry assay to measure antibody-dependent cellular cytotoxicity against SARS-CoV-2 Spike-expressing cells.

机构信息

Centre de Recherche du CHUM, Montréal, QC H2X 0A9, Canada.

Département de Microbiologie, Infectiologie et Immunologie, Université de Montréal, Montréal, QC H3T 1J4, Canada.

出版信息

STAR Protoc. 2021 Dec 17;2(4):100851. doi: 10.1016/j.xpro.2021.100851. Epub 2021 Sep 13.

Abstract

Antibodies can engage specific receptors at the surface of effector cells and mediate several functions beyond viral neutralization. Increasing evidence suggests that Fc-mediated effector functions, such as antibody-dependent cellular cytotoxicity (ADCC), have an important role in protection against severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infections. We engineered a cell line stably expressing a GFP-tagged SARS-CoV-2 spike to measure ADCC. This protocol provides an optimized way of measuring ADCC activity mediated by anti-SARS-CoV-2 Spike monoclonal antibodies or plasma from previously infected or vaccinated individuals. For complete details on the use and execution of this protocol, please refer to Anand et al. (2021b).

摘要

抗体可以与效应细胞表面的特定受体结合,并介导除中和病毒以外的多种功能。越来越多的证据表明,Fc 介导的效应功能,如抗体依赖性细胞毒性(ADCC),在预防严重急性呼吸综合征冠状病毒 2(SARS-CoV-2)感染方面起着重要作用。我们构建了一株稳定表达 GFP 标记的 SARS-CoV-2 刺突蛋白的细胞系,用于测量 ADCC。该方案提供了一种优化的方法来测量由抗 SARS-CoV-2 刺突单克隆抗体或来自先前感染或接种疫苗个体的血浆介导的 ADCC 活性。有关此方案的使用和执行的完整详细信息,请参阅 Anand 等人。(2021b)。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/66cd/8488403/8f337ef2ada0/fx1.jpg

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