Janssen Vaccines & Prevention, Archimedesweg 4-6, Leiden 2333, the Netherlands.
Vaccine Research Center, National Institute of Allergy and Infectious Diseases, NIH, Bethesda, MD 20892, USA.
Cell Rep. 2018 Apr 10;23(2):584-595. doi: 10.1016/j.celrep.2018.03.061.
The heavily glycosylated native-like envelope (Env) trimer of HIV-1 is expected to have low immunogenicity, whereas misfolded forms are often highly immunogenic. High-quality correctly folded Envs may therefore be critical for developing a vaccine that induces broadly neutralizing antibodies. Moreover, the high variability of Env may require immunizations with multiple Envs. Here, we report a universal strategy that provides for correctly folded Env trimers of high quality and yield through a repair-and-stabilize approach. In the repair stage, we utilized a consensus strategy that substituted rare strain-specific residues with more prevalent ones. The stabilization stage involved structure-based design and experimental assessment confirmed by crystallographic feedback. Regions important for the refolding of Env were targeted for stabilization. Notably, the α9-helix and an intersubunit β sheet proved to be critical for trimer stability. Our approach provides a means to produce prefusion-closed Env trimers from diverse HIV-1 strains, a substantial advance for vaccine development.
HIV-1 高度糖基化的天然样包膜 (Env) 三聚体预计免疫原性较低，而错误折叠的形式通常具有高度免疫原性。因此，高质量正确折叠的 Env 对于开发诱导广泛中和抗体的疫苗可能至关重要。此外，Env 的高度变异性可能需要用多种 Env 进行免疫接种。在这里，我们报告了一种通用策略，通过修复和稳定方法提供高质量和高产量的正确折叠的 Env 三聚体。在修复阶段，我们利用了一种共识策略，用更常见的取代稀有株特异性残基。稳定阶段涉及基于结构的设计，实验评估通过晶体学反馈得到证实。针对 Env 重折叠的重要区域进行了稳定化处理。值得注意的是，α9 螺旋和亚基间 β 片层被证明对三聚体稳定性至关重要。我们的方法提供了一种从多种 HIV-1 株生产预融合封闭 Env 三聚体的手段，这是疫苗开发的重大进展。
