Department of Chemistry and Biochemistry, University of Bern, Freiestrasse 3, 3012 Bern, Switzerland.
J Virol. 2010 Nov;84(22):11737-46. doi: 10.1128/JVI.01143-10. Epub 2010 Sep 8.
Globoside (Gb4Cer), Ku80 autoantigen, and α5β1 integrin have been identified as cell receptors/coreceptors for human parvovirus B19 (B19V), but their role and mechanism of interaction with the virus are largely unknown. In UT7/Epo cells, expression of Gb4Cer and CD49e (integrin alpha-5) was high, but expression of Ku80 was insignificant. B19V colocalized with Gb4Cer and, to a lesser extent, with CD49e. However, only anti-Gb4Cer antibodies could disturb virus attachment. Only a small proportion of cell-bound viruses were internalized, while the majority became detached from the receptor. When added to uninfected cells, the receptor-detached virus showed superior cell binding capacity and infectivity. Attachment of B19V to cells triggered conformational changes in the capsid leading to the accessibility of the N terminus of VP1 (VP1u) to antibodies, which was maintained in the receptor-detached virus. VP1u became similarly accessible to antibodies following incubation of B19V particles with increasing concentrations of purified Gb4Cer. The receptor-mediated exposure of VP1u is critical for virus internalization, since capsids lacking VP1 could bind to cells but were not internalized. Moreover, an antibody against the N terminus of VP1u disturbed virus internalization, but only when present during and not after virus attachment, indicating the involvement of this region in binding events required for internalization. These results suggest that Gb4Cer is not only the primary receptor for B19V attachment but also the mediator of capsid rearrangements required for subsequent interactions leading to virus internalization. The capacity of the virus to detach and reattach again would enhance the probability of productive infections.
糖鞘脂(Gb4Cer)、Ku80 自身抗原和 α5β1 整合素已被鉴定为人类细小病毒 B19(B19V)的细胞受体/辅助受体,但它们与病毒相互作用的作用和机制在很大程度上尚不清楚。在 UT7/Epo 细胞中,Gb4Cer 和 CD49e(整合素 α-5)的表达水平较高,但 Ku80 的表达水平不显著。B19V 与 Gb4Cer 共定位,与 CD49e 共定位程度较低。然而,只有抗 Gb4Cer 抗体才能干扰病毒附着。只有一小部分结合在细胞上的病毒被内化,而大部分则从受体上脱落。当添加到未感染的细胞中时,受体脱落的病毒表现出更高的细胞结合能力和感染力。B19V 附着到细胞上会引发衣壳构象变化,导致 VP1(VP1u)的 N 端易于与抗体结合,这种情况在受体脱落的病毒中得以维持。随着 B19V 颗粒与纯化的 Gb4Cer 浓度的增加,VP1u 对抗体的可及性增加。受体介导的 VP1u 暴露对于病毒内化至关重要,因为缺乏 VP1 的衣壳可以与细胞结合,但不能被内化。此外,针对 VP1u N 端的抗体干扰病毒内化,但仅在病毒附着期间且不在附着后存在时才会干扰病毒内化,表明该区域参与内化所需的结合事件。这些结果表明,Gb4Cer 不仅是 B19V 附着的主要受体,也是随后导致病毒内化的衣壳重排的中介。病毒脱落和再次附着的能力会增加有性感染的可能性。