Watkins Hannah C, Rappazzo C Garrett, Higgins Jaclyn S, Sun Xiangjie, Brock Nicole, Chau Annie, Misra Aditya, Cannizzo Joseph P B, King Michael R, Maines Taronna R, Leifer Cynthia A, Whittaker Gary R, DeLisa Matthew P, Putnam David
Meinig School of Biomedical Engineering, Cornell University, Ithaca, NY 14853, USA.
Department of Biological and Environmental Engineering, Cornell University, Ithaca, NY 14853 USA.
Mol Ther. 2017 Apr 5;25(4):989-1002. doi: 10.1016/j.ymthe.2017.01.010. Epub 2017 Feb 16.
Recombinant, Escherichia coli-derived outer membrane vesicles (rOMVs), which display heterologous protein subunits, have potential as a vaccine adjuvant platform. One drawback to rOMVs is their lipopolysaccharide (LPS) content, limiting their translatability to the clinic due to potential adverse effects. Here, we explore a unique rOMV construct with structurally remodeled lipids containing only the lipid IVa portion of LPS, which does not stimulate human TLR4. The rOMVs are derived from a genetically engineered B strain of E. coli, ClearColi, which produces lipid IVa, and which was further engineered in our laboratory to hypervesiculate and make rOMVs. We report that rOMVs derived from this lipid IVa strain have substantially attenuated pyrogenicity yet retain high levels of immunogenicity, promote dendritic cell maturation, and generate a balanced Th1/Th2 humoral response. Additionally, an influenza A virus matrix 2 protein-based antigen displayed on these rOMVs resulted in 100% survival against a lethal challenge with two influenza A virus strains (H1N1 and H3N2) in mice with different genetic backgrounds (BALB/c, C57BL/6, and DBA/2J). Additionally, a two-log reduction of lung viral titer was achieved in a ferret model of influenza infection with human pandemic H1N1. The rOMVs reported herein represent a potentially safe and simple subunit vaccine delivery platform.
展示异源蛋白亚基的重组大肠杆菌衍生外膜囊泡（rOMV）有潜力作为疫苗佐剂平台。rOMV的一个缺点是其脂多糖（LPS）含量，由于潜在的不良反应，限制了它们向临床的转化。在此，我们探索了一种独特的rOMV构建体，其脂质结构经过重塑，仅包含LPS的脂质IVa部分，该部分不会刺激人TLR4。rOMV来源于一种基因工程改造的大肠杆菌B菌株ClearColi，它能产生脂质IVa，并且在我们实验室中进一步改造以形成超囊泡并制备rOMV。我们报道，源自这种脂质IVa菌株的rOMV的致热原性已大幅减弱，但仍保留高水平的免疫原性，可促进树突状细胞成熟，并产生平衡的Th1/Th2体液反应。此外，展示在这些rOMV上的基于甲型流感病毒基质2蛋白的抗原，使具有不同遗传背景（BALB/c、C57BL/6和DBA/2J）的小鼠在受到两种甲型流感病毒株（H1N1和H3N2）的致死性攻击后100%存活。此外，在人类大流行H1N1流感感染的雪貂模型中，肺部病毒滴度降低了两个对数。本文报道的rOMV代表了一种潜在安全且简单的亚单位疫苗递送平台。
