Antibody Inhibition of Influenza A Virus Assembly and Release.
Antibody Inhibition of Influenza A Virus Assembly and Release.
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甲型流感病毒装配和释放的抗体抑制。
DOI：
10.1101/2023.08.08.552198
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发表时间：
2023
期刊：
影响因子：
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通讯作者：
Vahey,MichaelD
中科院分区：
文献类型：
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作者：
Yuanyuan,He;Zijian,Guo;Subiaur,Sofie;Benegal,Ananya;Vahey,MichaelD
抗体是流感病毒感染的前线防御者，通过多种互补机制提供保护。尽管已经显示单克隆抗体（mAb）的子集在病毒组装和释放水平上限制复制，但仍不清楚这种保护机制的有效性和普遍性，部分原因是将这种作用与抗体功能的其他方面分开的挑战。为了解决这个问题，我们开发了基于成像的测定，以确定针对IAV表面蛋白的广泛mAb如何有效地特异性限制病毒外出。我们发现，经典的抗血凝素中和抗体是广泛多功能的，抑制病毒组装和释放的浓度比它们抑制病毒进入的浓度高1-20倍。这些抗体也能够改变脱落病毒粒子的形态特征，减少丝状颗粒的比例。我们发现，针对神经氨酸酶和M2的抗体也限制了病毒的流出，并且抗神经氨酸酶单克隆抗体的抑制作用仅部分归因于酶活性的丧失。在所有情况下，抗原交联-无论是在感染细胞的表面上，病毒和细胞膜之间，或两者-在抑制中起着至关重要的作用，我们能够区分这些模式的实验和通过基于结构的计算模型。总之，这些结果提供了一个框架，解剖抗体的多功能性，可以帮助指导改进的治疗性抗体或疫苗的发展，并可以扩展到其他病毒家族和抗体isotypes.IMPORTANCEAntibodies针对甲型流感病毒提供多方面的保护，防止感染。虽然灵敏和定量测定被广泛用于测量病毒附着和进入的抑制，但不同抗体抑制病毒流出的能力尚不清楚。我们通过开发一种基于成像的方法来测量针对甲型流感病毒表面蛋白的一组单克隆抗体对病毒释放的抗体抑制来应对这一挑战。使用这种方法，我们发现抑制病毒外出是常见的，并且可以具有与抗体抑制病毒进入的能力相似的效力。深入了解抗体功能的这一未充分研究的方面可能有助于指导改进对策的发展。
Antibodies are frontline defenders against influenza virus infection, providing protection through multiple complementary mechanisms. Although a subset of monoclonal antibodies (mAbs) has been shown to restrict replication at the level of virus assembly and release, it remains unclear how potent and pervasive this mechanism of protection is, due in part to the challenge of separating this effect from other aspects of antibody function. To address this question, we developed imaging-based assays to determine how effectively a broad range of mAbs against the IAV surface proteins can specifically restrict viral egress. We find that classically neutralizing antibodies against hemagglutinin are broadly multifunctional, inhibiting virus assembly and release at concentrations 1–20-fold higher than the concentrations at which they inhibit viral entry. These antibodies are also capable of altering the morphological features of shed virions, reducing the proportion of filamentous particles. We find that antibodies against neuraminidase and M2 also restrict viral egress and that inhibition by anti-neuraminidase mAbs is only partly attributable to a loss in enzymatic activity. In all cases, antigen crosslinking—either on the surface of the infected cell, between the viral and cell membrane, or both—plays a critical role in inhibition, and we are able to distinguish between these modes experimentally and through a structure-based computational model. Together, these results provide a framework for dissecting antibody multifunctionality that could help guide the development of improved therapeutic antibodies or vaccines and that can be extended to other viral families and antibody isotypes.IMPORTANCEAntibodies against influenza A virus provide multifaceted protection against infection. Although sensitive and quantitative assays are widely used to measure inhibition of viral attachment and entry, the ability of diverse antibodies to inhibit viral egress is less clear. We address this challenge by developing an imaging-based approach to measure antibody inhibition of virus release across a panel of monoclonal antibodies targeting the influenza A virus surface proteins. Using this approach, we find that inhibition of viral egress is common and can have similar potency to the ability of an antibody to inhibit viral entry. Insights into this understudied aspect of antibody function may help guide the development of improved countermeasures.