Department of Chemistry, Vanderbilt University, Nashville, TN 37235.
Center for Structural Biology, Vanderbilt University, Nashville, TN 37235.
Proc Natl Acad Sci U S A. 2020 Dec 8;117(49):31142-31148. doi: 10.1073/pnas.1922654117. Epub 2020 Nov 23.
Marburg virus (MARV) disease is lethal, with fatality rates up to 90%. Neutralizing antibodies (Abs) are promising drug candidates to prevent or treat the disease. Current efforts are focused in part on vaccine development to induce such MARV-neutralizing Abs. We analyzed the antibody repertoire from healthy unexposed and previously MARV-infected individuals to assess if naïve repertoires contain suitable precursor antibodies that could become neutralizing with a limited set of somatic mutations. We computationally searched the human Ab variable gene repertoire for predicted structural homologs of the neutralizing Ab MR78 that is specific to the receptor binding site (RBS) of MARV glycoprotein (GP). Eight Ab heavy-chain complementarity determining region 3 (HCDR3) loops from MARV-naïve individuals and one from a previously MARV-infected individual were selected for testing as HCDR3 loop chimeras on the MR78 Ab framework. Three of these chimerized antibodies bound to MARV GP. We then tested a full-length native Ab heavy chain encoding the same 17-residue-long HCDR3 loop that bound to the MARV GP the best among the chimeric Abs tested. Despite only 57% amino acid sequence identity, the Ab from a MARV-naïve donor recognized MARV GP and possessed neutralizing activity against the virus. Crystallization of both chimeric and full-length native heavy chain-containing Abs provided structural insights into the mechanism of binding for these types of Abs. Our work suggests that the MARV GP RBS is a promising candidate for epitope-focused vaccine design to induce neutralizing Abs against MARV.
马尔堡病毒 (MARV) 病致死率高达 90%。中和抗体 (Abs) 是预防或治疗这种疾病的有前途的药物候选物。目前的努力部分集中在疫苗开发上,以诱导产生这种 MARV 中和 Abs。我们分析了健康未暴露和以前感染过 MARV 的个体的抗体库,以评估天然库中是否含有合适的前体抗体,这些抗体在经过有限数量的体细胞突变后可能成为中和抗体。我们通过计算搜索人类 Ab 可变基因库,寻找针对 MARV 糖蛋白 (GP) 受体结合位点 (RBS) 的中和 Ab MR78 的预测结构同源物。从 MARV 未感染个体中选择了 8 个 Ab 重链互补决定区 3 (HCDR3) 环和一个来自以前感染过 MARV 的个体的 HCDR3 环,作为 MR78 Ab 框架的 HCDR3 环嵌合体进行测试。其中 3 个嵌合抗体与 MARV GP 结合。然后,我们测试了一个全长的天然 Ab 重链,该重链编码与嵌合 Abs 中结合 MARV GP 最好的相同的 17 个残基长的 HCDR3 环。尽管只有 57%的氨基酸序列同一性,但来自 MARV 未感染供体的 Ab 识别 MARV GP 并具有中和该病毒的活性。嵌合和全长天然重链包含 Abs 的结晶提供了这些类型 Abs 结合机制的结构见解。我们的工作表明,MARV GP RBS 是针对表位的疫苗设计的有前途的候选物,可诱导针对 MARV 的中和 Abs。