Pugh J C, Di Q, Mason W S, Simmons H
Fox Chase Cancer Center, Philadelphia, Pennsylvania 19111, USA.
J Virol. 1995 Aug;69(8):4814-22. doi: 10.1128/JVI.69.8.4814-4822.1995.
To test the hypothesis that susceptibility of hepatocytes to duck hepatitis B virus (DHBV) infection requires cell surface receptors that bind the virus in a specific manner, we developed an assay for the binding of DHBV particles to monolayers of intact cells, using radiolabeled immunoglobulin G specific for DHBV envelope protein. Both noninfectious DHBV surface antigen particles and infectious virions bound to a susceptible fraction (approximately 60%) of Pekin duck hepatocytes. In contrast, binding did not occur to cells that were not susceptible to DHBV infection, including Pekin duck fibroblasts and chicken hepatocytes, and binding to Muscovy duck hepatocytes, which are only weakly susceptible (approximately 1% of cells) to DHBV infection, was virtually undetectable. Within a monolayer, individual Pekin duck hepatocytes appeared to differ markedly in the capacity to bind DHBV, which may explain difficulties that have been encountered in infecting 100% of cells in culture. We have also found that the loss of susceptibility to infection with DHBV that occurs when Pekin duck hepatocytes are maintained for more than a few days in culture correlates with a decline in the number of cells that bind virus particles efficiently. All of these results support the interpretation that the binding event detected by our assay is associated with the interaction between DHBV and specific cell surface receptors that are required for initiation of infection. Our assay may facilitate isolation and identification of hepatocyte receptors for this virus.
为了验证肝细胞对鸭乙型肝炎病毒(DHBV)感染的易感性需要以特定方式结合该病毒的细胞表面受体这一假说,我们开发了一种检测方法,利用针对DHBV包膜蛋白的放射性标记免疫球蛋白G,来检测DHBV颗粒与完整细胞单层的结合。非感染性DHBV表面抗原颗粒和感染性病毒粒子均能与北京鸭肝细胞的一个易感部分(约60%)结合。相比之下,与对DHBV感染不易感的细胞(包括北京鸭成纤维细胞和鸡肝细胞)则不会发生结合,并且与仅对DHBV感染有微弱易感性(约1%的细胞)的番鸭肝细胞的结合实际上无法检测到。在一个单层内,单个北京鸭肝细胞在结合DHBV的能力上似乎存在显著差异,这可能解释了在培养中感染100%细胞时所遇到的困难。我们还发现,当北京鸭肝细胞在培养中维持超过几天时,其对DHBV感染的易感性丧失与有效结合病毒颗粒的细胞数量下降相关。所有这些结果都支持这样一种解释,即我们检测到的结合事件与DHBV和感染起始所需的特定细胞表面受体之间的相互作用有关。我们的检测方法可能有助于分离和鉴定该病毒的肝细胞受体。