Thompson Bruce S, Moesker Bastiaan, Smit Jolanda M, Wilschut Jan, Diamond Michael S, Fremont Daved H
Pathology and Immunology, Washington University School of Medicine, St Louis, MO, USA.
PLoS Pathog. 2009 May;5(5):e1000453. doi: 10.1371/journal.ppat.1000453. Epub 2009 May 29.
Defining the precise cellular mechanisms of neutralization by potently inhibitory antibodies is important for understanding how the immune system successfully limits viral infections. We recently described a potently inhibitory monoclonal antibody (MAb E16) against the envelope (E) protein of West Nile virus (WNV) that neutralizes infection even after virus has spread to the central nervous system. Herein, we define its mechanism of inhibition. E16 blocks infection primarily at a post-attachment step as antibody-opsonized WNV enters permissive cells but cannot escape from endocytic compartments. These cellular experiments suggest that E16 blocks the acid-catalyzed fusion step that is required for nucleocapsid entry into the cytoplasm. Indeed, E16 directly inhibits fusion of WNV with liposomes. Additionally, low-pH exposure of E16-WNV complexes in the absence of target membranes did not fully inactivate infectious virus, further suggesting that E16 prevents a structural transition required for fusion. Thus, a strongly neutralizing anti-WNV MAb with therapeutic potential is potently inhibitory because it blocks viral fusion and thereby promotes clearance by delivering virus to the lysosome for destruction.
明确强效抑制性抗体中和作用的确切细胞机制,对于理解免疫系统如何成功限制病毒感染至关重要。我们最近描述了一种针对西尼罗河病毒(WNV)包膜(E)蛋白的强效抑制性单克隆抗体(MAb E16),即使在病毒扩散至中枢神经系统后,该抗体仍能中和感染。在此,我们确定了其抑制机制。E16主要在附着后步骤阻断感染,因为抗体调理的WNV进入允许性细胞后无法从内吞小室逃逸。这些细胞实验表明,E16阻断了核衣壳进入细胞质所需的酸催化融合步骤。事实上,E16直接抑制WNV与脂质体的融合。此外,在没有靶膜的情况下,E16-WNV复合物暴露于低pH环境并未使感染性病毒完全失活,这进一步表明E16阻止了融合所需的结构转变。因此,一种具有治疗潜力的强效中和抗WNV单克隆抗体具有强效抑制作用,因为它阻断病毒融合,从而通过将病毒递送至溶酶体进行破坏来促进病毒清除。