Carr Caleb R, Crawford Katharine H D, Murphy Michael, Galloway Jared G, Haddox Hugh K, Matsen Frederick A, Andersen Kristian G, King Neil P, Bloom Jesse D
Basic Sciences Division and Computational Biology Program, Fred Hutchinson Cancer Center, Seattle, WA 98109, USA.
Department of Genome Sciences, University of Washington, Seattle, WA 98109, USA.
bioRxiv. 2024 Feb 6:2024.02.05.579020. doi: 10.1101/2024.02.05.579020.
Lassa virus is estimated to cause thousands of human deaths per year, primarily due to spillovers from its natural host, rodents. Efforts to create vaccines and antibody therapeutics must account for the evolutionary variability of 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 use pseudovirus deep mutational scanning to measure how nearly all GPC amino-acid mutations affect cell entry and antibody neutralization. Our experiments define functional constraints throughout GPC. We quantify how GPC mutations affect neutralization by a panel of monoclonal antibodies and show that all antibodies are escaped by mutations that exist 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 design of therapeutics and vaccines.
据估计，拉沙病毒每年导致数千人死亡，主要原因是该病毒从其自然宿主啮齿动物溢出。研发疫苗和抗体疗法的工作必须考虑到拉沙病毒糖蛋白复合体（GPC）的进化变异性，该复合体介导病毒进入细胞，并且是中和抗体的靶标。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测量几乎所有GPC氨基酸突变如何影响细胞进入和抗体中和。我们的实验确定了整个GPC的功能限制。我们量化了GPC突变如何影响一组单克隆抗体的中和作用，并表明所有抗体都会被自然拉沙病毒谱系中存在的突变所逃逸。总体而言，我们的工作描述了一种生物安全2级方法，以阐明GPC可及的突变空间，并展示了对抗原变异的前瞻性表征如何有助于治疗药物和疫苗的设计。
