Functional genomics screens reveal a role for TBC1D24 and SV2B in antibody-dependent enhancement of dengue virus infection.

IF 4 2区 医学 Q2 VIROLOGY
Journal of Virology Pub Date : 2024-11-19 Epub Date: 2024-10-08 DOI:10.1128/jvi.01582-24
Laura Belmont, Maya Contreras, Catiana H Cartwright-Acar, Caleb D Marceau, Aditi Agrawal, Lisa M Levoir, Jay Lubow, Leslie Goo
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引用次数: 0

Abstract

Under some conditions, dengue virus (DENV) can hijack IgG antibodies to facilitate its uptake into target cells expressing Fc gamma receptors (FcgR)-a process known as antibody-dependent enhancement (ADE) of infection. Beyond a requirement for FcgR, host dependency factors for this unusual IgG-mediated infection route remain unknown. To identify cellular factors exclusively required for ADE, here, we performed CRISPR knockout (KO) screens in an in vitro system poorly permissive to infection in the absence of IgG antibodies. Validating our approach, a top hit was FcgRIIa, which facilitates the binding and internalization of IgG-bound DENV but is not required for canonical infection. Additionally, we identified host factors with no previously described role in DENV infection, including TBC1D24 and SV2B, which have known functions in regulated secretion. Using genetic knockout and trans-complemented cells, we validated a functional requirement for these host factors in ADE assays performed with monoclonal antibodies and polyclonal sera in multiple cell lines and using all four DENV serotypes. We show that knockout of TBC1D24 or SV2B impaired the binding of IgG-DENV complexes to cells without affecting FcgRIIa expression levels. Thus, we identify cellular factors beyond FcgR that promote efficient ADE of DENV infection. Our findings represent a first step toward advancing fundamental knowledge behind the biology of a non-canonical infection route implicated in disease.IMPORTANCEAntibodies can paradoxically enhance rather than inhibit dengue virus (DENV) infection in some cases. To advance knowledge of the functional requirements of antibody-dependent enhancement (ADE) of infection beyond existing descriptive studies, we performed a genome-scale CRISPR knockout (KO) screen in an optimized in vitro system permissive to efficient DENV infection only in the presence of IgG. In addition to FcgRIIa, a known receptor that facilitates IgG-mediated uptake of IgG-bound, but not naked DENV particles, our screens identified TBC1D24 and SV2B, cellular factors with no known role in DENV infection. We validated a functional role for TBC1D24 and SV2B in mediating ADE of all four DENV serotypes in different cell lines and using various antibodies. Thus, we identify cellular factors beyond Fc gamma receptors that promote ADE mechanisms. This study represents a first step toward advancing fundamental knowledge beyond a poorly understood non-canonical viral entry mechanism.

功能基因组学筛选揭示了 TBC1D24 和 SV2B 在抗体依赖性增强登革热病毒感染中的作用。
在某些条件下,登革热病毒(DENV)可以劫持 IgG 抗体以促进其被表达 Fc γ 受体(FcgR)的靶细胞吸收--这一过程被称为抗体依赖性感染增强(ADE)。除了对 FcgR 的要求外,这种不寻常的 IgG 介导的感染途径的宿主依赖性因素仍然未知。为了确定 ADE 专门需要的细胞因子,我们在缺乏 IgG 抗体的情况下对感染容许度较低的体外系统进行了 CRISPR 基因敲除(KO)筛选。FcgRIIa验证了我们的方法,它促进了IgG结合的DENV的结合和内化,但并非典型感染所必需。此外,我们还发现了以前没有描述过的在 DENV 感染中发挥作用的宿主因子,包括 TBC1D24 和 SV2B,它们在调节分泌方面具有已知的功能。利用基因敲除和反式补体细胞,我们在使用单克隆抗体和多克隆血清对多个细胞系进行的ADE检测中验证了这些宿主因子的功能要求,并使用了所有四种DENV血清型。我们发现,敲除 TBC1D24 或 SV2B 会影响 IgG-DENV 复合物与细胞的结合,但不会影响 FcgRIIa 的表达水平。因此,我们发现了FcgR之外促进DENV感染高效ADE的细胞因素。我们的研究结果代表了向推进与疾病相关的非典型感染途径生物学背后的基础知识迈出的第一步。重要意义抗体在某些情况下会自相矛盾地增强而不是抑制登革病毒(DENV)感染。为了在现有的描述性研究之外进一步了解抗体依赖性感染增强(ADE)的功能要求,我们在一个优化的体外系统中进行了基因组规模的CRISPR基因敲除(KO)筛选,该系统只有在IgG存在时才允许登革热病毒的有效感染。FcgRIIa是一种已知的受体,可促进IgG介导的与IgG结合的DENV颗粒的摄取,但不能促进裸体DENV颗粒的摄取,除了FcgRIIa,我们的筛选还发现了TBC1D24和SV2B,它们是在DENV感染中没有已知作用的细胞因子。我们在不同的细胞系中使用各种抗体验证了TBC1D24和SV2B在介导所有四种DENV血清型的ADE中的功能性作用。因此,我们确定了促进 ADE 机制的 Fc γ 受体之外的细胞因素。这项研究标志着我们迈出了第一步,在人们对非典型病毒进入机制了解甚少的基础上又向前迈进了一步。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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来源期刊
Journal of Virology
Journal of Virology 医学-病毒学
CiteScore
10.10
自引率
7.40%
发文量
906
审稿时长
1 months
期刊介绍: Journal of Virology (JVI) explores the nature of the viruses of animals, archaea, bacteria, fungi, plants, and protozoa. We welcome papers on virion structure and assembly, viral genome replication and regulation of gene expression, genetic diversity and evolution, virus-cell interactions, cellular responses to infection, transformation and oncogenesis, gene delivery, viral pathogenesis and immunity, and vaccines and antiviral agents.
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