Characterization of SARS-CoV-2 replication in human H1299/ACE2 cells: A versatile and practical infection model for antiviral research and beyond

IF 4.5 2区 医学 Q1 PHARMACOLOGY & PHARMACY
Clarisse Salgado-Benvindo , Ali Tas , Jessika C. Zevenhoven-Dobbe , Yvonne van der Meer , Igor A. Sidorov , Anouk A. Leijs , Patrick Wanningen , Anne T. Gelderloos , Puck B. van Kasteren , Eric J. Snijder , Martijn J. van Hemert
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引用次数: 0

Abstract

A range of cell culture infection models have been used to study SARS-CoV-2 and perform antiviral drug research. Commonly used African green monkey Vero, human lung-derived Calu-3 and ACE2+TMPRSS2-expressing A549 cells, each have their limitations. Here, we describe human ACE2-expressing H1299 lung cells as a more efficient and robust model for SARS-CoV-2 research. These cells are as easy to handle as Vero cells, support SARS-CoV-2 replication to high titers, display a functional innate immune response and are suitable for plaque assays, microscopy, the production of (genetically stable) virus stocks and antiviral assays. H1299/ACE2-based (CPE reduction) assays can be performed without adding a P-gP drug efflux pump inhibitor, which is often required in Vero-based assays. Moreover, H1299/ACE2 cells allowed us to perform CPE reduction assays with omicron variants that did not work in Vero-based assays. In summary, H1299/ACE2 cells are a versatile infection model to study SARS-CoV-2 replication in the context of antiviral drug development and virus-host interaction studies.

SARS-CoV-2 在人类 H1299/ACE2 细胞中复制的特征:一种用于抗病毒研究及其他领域的多功能实用感染模型。
在研究 SARS-CoV-2 和进行抗病毒药物研究时,使用了一系列细胞培养感染模型。常用的非洲绿猴 Vero、人肺源 Calu-3 和 ACE2+TMPRSS2 表达的 A549 细胞各有其局限性。在这里,我们描述了表达人 ACE2 的 H1299 肺细胞,它是研究 SARS-CoV-2 的一种更有效、更强大的模型。这些细胞与 Vero 细胞一样易于处理,支持高滴度的 SARS-CoV-2 复制,显示出功能性先天免疫反应,适用于斑块检测、显微镜检查、生产(基因稳定的)病毒储备和抗病毒检测。基于 H1299/ACE2(CPE 减少)的检测可以在不添加 P-gP 药物外排泵抑制剂的情况下进行,而在基于 Vero 的检测中通常需要添加 P-gP 药物外排泵抑制剂。此外,H1299/ACE2 细胞还能让我们用在基于 Vero 的试验中不起作用的欧米克隆变体进行 CPE 还原试验。总之,H1299/ACE2 细胞是在抗病毒药物开发和病毒-宿主相互作用研究中研究 SARS-CoV-2 复制的多功能感染模型。
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来源期刊
Antiviral research
Antiviral research 医学-病毒学
CiteScore
17.10
自引率
3.90%
发文量
157
审稿时长
34 days
期刊介绍: Antiviral Research is a journal that focuses on various aspects of controlling viral infections in both humans and animals. It is a platform for publishing research reports, short communications, review articles, and commentaries. The journal covers a wide range of topics including antiviral drugs, antibodies, and host-response modifiers. These topics encompass their synthesis, in vitro and in vivo testing, as well as mechanisms of action. Additionally, the journal also publishes studies on the development of new or improved vaccines against viral infections in humans. It delves into assessing the safety of drugs and vaccines, tracking the evolution of drug or vaccine-resistant viruses, and developing effective countermeasures. Another area of interest includes the identification and validation of new drug targets. The journal further explores laboratory animal models of viral diseases, investigates the pathogenesis of viral diseases, and examines the mechanisms by which viruses avoid host immune responses.
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