Redox-triggered infection by disulfide-shackled human immunodeficiency virus type 1 pseudovirions
Redox-triggered infection by disulfide-shackled human immunodeficiency virus type 1 pseudovirions
复制标题
DOI：
10.1128/jvi.77.10.5678-5684.2003
复制
发表时间：
2003-05-01
影响因子：
5.4
通讯作者：
Burton, DR
中科院分区：
文献类型：
--
作者：
Binley, JM;Cayanan, CS;Burton, DR
我们先前描述了一种1型人类免疫缺陷病毒（HIV - 1）包膜突变体，它在gp120表面蛋白和gp41跨膜蛋白之间引入了一个二硫键（J. M. 宾利，R. W. 桑德斯，B. 克拉斯，N. 舒尔克，A. 马斯特，Y. 郭，F. 卡朱莫，D. J. 安塞尔马，P. J. 马登，W. C. 奥尔森和J. P. 摩尔，《病毒学杂志》74：627 - 643，2000年）。在此，我们制备了携带突变包膜和报告基因的假病毒粒子，以研究该突变体的感染特性。这些假病毒粒子会附着在表达CD4和辅助受体的细胞上，但只有在还原剂触发时才会感染，这意味着gp120 - gp41的解离对于感染是必需的。进一步的研究表明，在CD4和辅助受体结合后病毒进入过程受阻。通过测量各种进入抑制剂对受阻中间体的活性，我们发现针对gp120的抑制剂通常在病毒附着之前起作用，而gp41抑制剂能够在附着之后起作用。出乎意料的是，HIV - 1阳性血清中的很大一部分抗体在病毒附着之后中和病毒，这表明下游融合事件和结构在宿主免疫反应中起重要作用。总体而言，这种被二硫键束缚的病毒是一种独特的工具，在疫苗设计、药物研发以及阐明HIV - 1进入过程方面具有潜在的用途。
We previously described a human immunodeficiency virus type 1 (HIV-1) envelope mutant that introduces a disulfide bridge between the gp120 surface proteins and gp41 transmembrane proteins (J. M. Binley, R. W. Sanders, B. Clas, N. Schuelke, A. Master, Y. Guo, F. Kajumo, D. J. Anselma, P. J. Maddon, W. C. Olson, and J. P. Moore, J. Virol. 74:627-643, 2000). Here we produced pseudovirions bearing the mutant envelope and a reporter gene to examine the mutant's infectious properties. These pseudovirions attach to cells expressing CD4 and coreceptor but infect only when triggered with reducing agent, implying that gp120-gp41 dissociation is necessary for infection. Further studies suggested that virus entry was arrested after CD4 and coreceptor engagement. By measuring the activities of various entry inhibitors against the arrested intermediate, we found that gp120-targeting inhibitors typically act prior to virus attachment, whereas gp41 inhibitors are able to act postattachment. Unexpectedly, a significant fraction of antibodies in HIV-1-positive sera neutralized virus postattachment, suggesting that downstream fusion events and structures figure prominently in the host immune response. Overall, this disulfide-shackled virus is a unique tool with potential utility in vaccine design, drug discovery, and elucidation of the HIV-1 entry process.