Basic Sciences Division and Computational Biology Program, Fred Hutchinson Cancer Center, Seattle, WA 98109, USA; Department of Genome Sciences, University of Washington, Seattle, WA 98195, USA; Medical Scientist Training Program, University of Washington, Seattle, WA 98195, USA.
Department of Biochemistry, University of Washington, Seattle, WA 98195, USA; Institute for Protein Design, University of Washington, Seattle, WA 98195, USA.
Immunity. 2024 Sep 10;57(9):2061-2076.e11. doi: 10.1016/j.immuni.2024.06.013. Epub 2024 Jul 15.
Lassa virus is estimated to cause thousands of human deaths per year, primarily due to spillovers from its natural host, Mastomys rodents. Efforts to create vaccines and antibody therapeutics must account for the evolutionary variability of the Lassa virus's glycoprotein complex (GPC), which mediates viral entry into cells and is the target of neutralizing antibodies. To map the evolutionary space accessible to GPC, we used pseudovirus deep mutational scanning to measure how nearly all GPC amino-acid mutations affected cell entry and antibody neutralization. Our experiments defined functional constraints throughout GPC. We quantified how GPC mutations affected neutralization with a panel of monoclonal antibodies. All antibodies tested were escaped by mutations that existed among natural Lassa virus lineages. Overall, our work describes a biosafety-level-2 method to elucidate the mutational space accessible to GPC and shows how prospective characterization of antigenic variation could aid the design of therapeutics and vaccines.
拉沙病毒估计每年导致数千人死亡，主要是由于其自然宿主——巨鼠科的啮齿动物的溢出。为了开发疫苗和抗体治疗药物，必须考虑到拉沙病毒糖蛋白复合物（GPC）的进化可变性，该复合物介导病毒进入细胞，是中和抗体的靶标。为了绘制 GPC 可及的进化空间，我们使用假病毒深度突变扫描来衡量 GPC 的几乎所有氨基酸突变如何影响细胞进入和抗体中和。我们的实验在整个 GPC 中定义了功能限制。我们使用单克隆抗体对 GPC 突变如何影响中和作用进行了定量。测试的所有抗体都被天然拉沙病毒谱系中存在的突变所逃逸。总的来说，我们的工作描述了一种生物安全 2 级方法来阐明 GPC 可及的突变空间，并展示了抗原变异的前瞻性特征如何有助于治疗药物和疫苗的设计。
