Bai Xiaohui, Li Chenyang, Qiu Jiahui, Wu Leyang, Liu Xinqi, Yin Te, Jin Li, Hua Zichun
The State Key Laboratory of Pharmaceutical Biotechnology and Department of Neurology of Nanjing Drum Tower Hospital, School of Life Sciences and The Affiliated Hospital of Nanjing University Medical School, Nanjing University, Nanjing 210023, China.
The State Key Laboratory of Pharmaceutical Biotechnology and Department of Neurology of Nanjing Drum Tower Hospital, School of Life Sciences and The Affiliated Hospital of Nanjing University Medical School, Nanjing University, Nanjing 210023, China; Changzhou High-Tech Research Institute of Nanjing University and Jiangsu TargetPharma Laboratories Inc, Changzhou 213164, China.
J Control Release. 2025 Feb 10;378:687-700. doi: 10.1016/j.jconrel.2024.12.022. Epub 2024 Dec 26.
As natural nanoparticle, the bacterial outer membrane vesicles (OMV) hold great potential in protein vaccines because of its self-adjuvant properties and good biocompatibility. However, the inherent immunotoxicity seriously hampers the application of OMV as protein antigens delivery carrier. Here, an attenuated OMV was constructed by elimination of the flagella protein from its surface and removal of the phosphate group of LPS at position one via gene-editing strategy. The gene-edited outer membrane vesicles (EMV) effectively reduced the levels of pro-inflammatory factors TNF-α and IL-6 in mouse blood by at least 10-fold and 15-fold respectively, compared to wild type OMV (WT-OMV). Importantly, protein antigens are conveniently displayed on EMV by employing a plug-and-display procedure, whereby the exterior of biotinylated EMV can be readily decorated with a synthetic protein comprised of target antigen fused to a biotin-binding protein. EMV greatly increased the uptake of antigen by dendritic cells (DCs) and promoted their maturation. EMV-antigen complex induces a robust antigen-specific antibody responses and cellular immune responses. We propose that EMV have great potential as protein antigens delivery vehicle for preventing different infectious diseases.
作为天然纳米颗粒，细菌外膜囊泡（OMV）因其自身佐剂特性和良好的生物相容性，在蛋白质疫苗方面具有巨大潜力。然而，其固有的免疫毒性严重阻碍了OMV作为蛋白质抗原递送载体的应用。在此，通过基因编辑策略从其表面消除鞭毛蛋白并去除脂多糖（LPS）第一位的磷酸基团，构建了减毒OMV。与野生型OMV（WT-OMV）相比，基因编辑后的外膜囊泡（EMV）可有效降低小鼠血液中促炎因子TNF-α和IL-6的水平，分别至少降低10倍和15倍。重要的是，通过采用即插即显方法，可方便地在EMV上展示蛋白质抗原，由此生物素化的EMV外部可轻松用由与生物素结合蛋白融合的靶抗原组成的合成蛋白进行修饰。EMV大大增加了树突状细胞（DCs）对抗原的摄取并促进其成熟。EMV-抗原复合物诱导强烈的抗原特异性抗体反应和细胞免疫反应。我们认为，EMV作为预防不同传染病的蛋白质抗原递送载体具有巨大潜力。
