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针对糖蛋白B融合结构域的抗体中和单纯疱疹病毒的机制。

Mechanism of neutralization of herpes simplex virus by antibodies directed at the fusion domain of glycoprotein B.

作者信息

Cairns Tina M, Fontana Juan, Huang Zhen-Yu, Whitbeck J Charles, Atanasiu Doina, Rao Samhita, Shelly Spencer S, Lou Huan, Ponce de Leon Manuel, Steven Alasdair C, Eisenberg Roselyn J, Cohen Gary H

机构信息

Department of Microbiology, School of Dental Medicine, University of Pennsylvania, Philadelphia, Pennsylvania, USA.

出版信息

J Virol. 2014 Mar;88(5):2677-89. doi: 10.1128/JVI.03200-13. Epub 2013 Dec 18.

Abstract

UNLABELLED

Glycoprotein B (gB), the fusogen of herpes simplex virus (HSV), is a class III fusion protein with a trimeric ectodomain of known structure for the postfusion state. Seen by negative-staining electron microscopy, it presents as a rod with three lobes (base, middle, and crown). gB has four functional regions (FR), defined by the physical location of epitopes recognized by anti-gB neutralizing monoclonal antibodies (MAbs). Located in the base, FR1 contains two internal fusion loops (FLs) and is the site of gB-lipid interaction (the fusion domain). Many of the MAbs to FR1 are neutralizing, block cell-cell fusion, and prevent the association of gB with lipid, suggesting that these MAbs affect FL function. Here we characterize FR1 epitopes by using electron microscopy to visualize purified Fab-gB ectodomain complexes, thus confirming the locations of several epitopes and localizing those of MAbs DL16 and SS63. We also generated MAb-resistant viruses in order to localize the SS55 epitope precisely. Because none of the epitopes of our anti-FR1 MAbs mapped to the FLs, we hyperimmunized rabbits with FL1 or FL2 peptides to generate polyclonal antibodies (PAbs). While the anti-FL1 PAb failed to bind gB, the anti-FL2 PAb had neutralizing activity, implying that the FLs become exposed during virus entry. Unexpectedly, the anti-FL2 PAb (and the anti-FR1 MAbs) bound to liposome-associated gB, suggesting that their epitopes are accessible even when the FLs engage lipid. These studies provide possible mechanisms of action for HSV neutralization and insight into how gB FR1 contributes to viral fusion.

IMPORTANCE

For herpesviruses, such as HSV, entry into a target cell involves transfer of the capsid-encased genome of the virus to the target cell after fusion of the lipid envelope of the virus with a lipid membrane of the host. Virus-encoded glycoproteins in the envelope are responsible for fusion. Antibodies to these glycoproteins are important biological tools, providing a way of examining how fusion works. Here we used electron microscopy and other techniques to study a panel of anti-gB antibodies. Some, with virus-neutralizing activity, impair gB-lipid association. We also generated a peptide antibody against one of the gB fusion loops; its properties provide insight into the way the fusion loops function as gB transits from its prefusion form to an active fusogen.

摘要

未标记

糖蛋白B(gB)是单纯疱疹病毒(HSV)的融合蛋白,属于III类融合蛋白,其三聚体外结构域在融合后状态下具有已知结构。通过负染色电子显微镜观察,它呈现为带有三个叶(基部、中部和冠部)的杆状结构。gB有四个功能区(FR),由抗gB中和单克隆抗体(MAb)识别的表位的物理位置定义。FR1位于基部,包含两个内部融合环(FL),是gB与脂质相互作用的位点(融合结构域)。许多针对FR1的MAb具有中和作用,可阻断细胞间融合,并阻止gB与脂质结合,这表明这些MAb影响FL的功能。在这里,我们通过电子显微镜观察纯化的Fab-gB外结构域复合物来表征FR1表位,从而确认了几个表位的位置,并确定了MAb DL16和SS63的表位位置。我们还产生了抗MAb病毒,以便精确确定SS55表位的位置。由于我们的抗FR1 MAb的表位均未映射到FL,我们用FL1或FL2肽对兔子进行超免疫以产生多克隆抗体(PAb)。虽然抗FL1 PAb未能结合gB,但抗FL2 PAb具有中和活性,这意味着FL在病毒进入过程中会暴露出来。出乎意料的是,抗FL2 PAb(以及抗FRl MAb)与脂质体相关的gB结合,这表明即使FL与脂质结合,它们的表位也是可及的。这些研究为HSV中和作用提供了可能的作用机制,并深入了解了gB FR1如何促进病毒融合。

重要性

对于疱疹病毒,如HSV,进入靶细胞涉及病毒的衣壳包裹基因组在病毒的脂质包膜与宿主的脂质膜融合后转移到靶细胞。包膜中病毒编码的糖蛋白负责融合。针对这些糖蛋白的抗体是重要的生物学工具,提供了一种研究融合如何发生的方法。在这里,我们使用电子显微镜和其他技术研究了一组抗gB抗体。一些具有病毒中和活性的抗体损害gB与脂质的结合。我们还产生了一种针对gB融合环之一的肽抗体;其特性为融合环在gB从其融合前形式转变为活性融合蛋白过程中的功能方式提供了深入了解。

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本文引用的文献

3
Structure of RSV fusion glycoprotein trimer bound to a prefusion-specific neutralizing antibody.
Science. 2013 May 31;340(6136):1113-7. doi: 10.1126/science.1234914. Epub 2013 Apr 25.
4
Extensive mutagenesis of the HSV-1 gB ectodomain reveals remarkable stability of its postfusion form.
J Mol Biol. 2013 Jun 12;425(11):2056-2071. doi: 10.1016/j.jmb.2013.03.001. Epub 2013 Mar 13.
7
HCMV gB shares structural and functional properties with gB proteins from other herpesviruses.
Virology. 2013 Jan 20;435(2):239-49. doi: 10.1016/j.virol.2012.09.024. Epub 2012 Oct 22.
10
Capturing the herpes simplex virus core fusion complex (gB-gH/gL) in an acidic environment.
J Virol. 2011 Jul;85(13):6175-84. doi: 10.1128/JVI.00119-11. Epub 2011 Apr 20.

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