KJ Biosciences LLC, 2151 Harvey Mitchell Parkway, Suite 306, College Station, Texas, United States.
KJ Biosciences LLC, 2151 Harvey Mitchell Parkway, Suite 306, College Station, Texas, United States.
Vaccine. 2017 Dec 15;35(50):7026-7032. doi: 10.1016/j.vaccine.2017.10.051. Epub 2017 Nov 6.
A highly effective antigen construct for presenting conserved antigen domains is essential to the development of a universal influenza vaccine. We have developed a novel dual-domain nanoparticle fusion protein (DDNFP) which allows independent presentation of two conserved domains. The conserved domains used were from two separate viral surface proteins, M2e of M2 and fusion peptide (FP) or long alpha helix (CD) of HA2. The carrier is a novel nanoparticle protein - the dodecameric DNA binding protein from starved cells (Dps) of bacteria or archaea. Dps was found to be uniquely capable of simultaneous fusion and surface presentation at both N- and C-termini while retaining the ability to form nanoparticles. Thus, DDNFPs with M2e and FP or CD fused at N- and C-termini of Dps from E. coli (EcDps) or other bacteria were first constructed based on the H1 subtype sequences along with corresponding single-domain nanoparticle fusion proteins (SDNFPs). They were expressed at high levels in bacteria and found to form nanoparticles of the expected size (∼9 nm). They were stable against treatment at high temperatures. The DDNFPs (M2e-EcDps-FP and M2e-EcDps-CD) induced strong antibody responses against individual antigen domains and provided full protection against lethal challenge with PR8 virus (H1N1). Importantly, the protection by DDNFPs was synergistically enhanced as compared to SDNFPs. The M2e-EcDps-CD provided an even stronger protection than M2e-EcDps-FP and therefore appeared to be the superior construct. Together, with novel domain combination, enhanced protection and ease of production, this M2e/CD DDNFP could potentially be a highly effective antigen construct for the universal influenza vaccine.
一种能够有效展示保守抗原结构域的高效抗原构建物对于开发通用流感疫苗至关重要。我们开发了一种新型的双结构域纳米颗粒融合蛋白（DDNFP），该蛋白可独立展示两种保守结构域。所使用的保守结构域来自两种不同的病毒表面蛋白，M2 的 M2e 和融合肽（FP）或 HA2 的长α螺旋（CD）。载体是一种新型的纳米颗粒蛋白——细菌或古菌饥饿细胞中的十二聚体 DNA 结合蛋白（Dps）。研究发现，Dps 具有独特的在 N 端和 C 端同时融合和表面呈现的能力，同时保留形成纳米颗粒的能力。因此，根据 H1 亚型序列以及相应的单结构域纳米颗粒融合蛋白（SDNFP），首次构建了在大肠杆菌（EcDps）或其他细菌的 Dps 的 N 端和 C 端融合有 M2e 和 FP 或 CD 的 DDNFP。它们在细菌中高水平表达，并发现它们形成了预期大小（约 9nm）的纳米颗粒。它们在高温处理下稳定。DDNFP（M2e-EcDps-FP 和 M2e-EcDps-CD）诱导针对单个抗原结构域的强烈抗体反应，并为 PR8 病毒（H1N1）的致死性攻击提供完全保护。重要的是，与 SDNFP 相比，DDNFP 的保护作用呈协同增强。M2e-EcDps-CD 提供了比 M2e-EcDps-FP 更强的保护作用，因此似乎是更好的构建体。综上所述，这种 M2e/CD DDNFP 具有新颖的结构域组合、增强的保护作用和易于生产的特点，有可能成为一种高效的通用流感疫苗抗原构建物。
