Chung Tai-Chun, Jones Charles H, Gollakota Akhila, Kamal Ahmadi Mahmoud, Rane Snehal, Zhang Guojian, Pfeifer Blaine A
Department of Chemical and Biological Engineering, University at Buffalo, State University of New York, Buffalo, New York 14260-4200, United States.
Mol Pharm. 2015 May 4;12(5):1691-700. doi: 10.1021/acs.molpharmaceut.5b00172. Epub 2015 Apr 16.
Bactofection offers a gene delivery option particularly useful in the context of immune modulation. The bacterial host naturally attracts recognition and cellular uptake by antigen presenting cells (APCs) as the initial step in triggering an immune response. Moreover, depending on the bacterial vector, molecular biology tools are available to influence and/or overcome additional steps and barriers to effective antigen presentation. In this work, molecular engineering was applied using Escherichia coli as a bactofection vector. In particular, the bacteriophage ΦX174 lysis E (LyE) gene was designed for variable expression across strains containing different levels of lysteriolysin O (LLO). The objective was to generate a bacterial vector with improved attenuation and delivery characteristics. The resulting strains exhibited enhanced gene and protein release and inducible cellular death. In addition, the new vectors demonstrated improved gene delivery and cytotoxicity profiles to RAW264.7 macrophage APCs.
细菌转染提供了一种在免疫调节背景下特别有用的基因递送选择。作为触发免疫反应的第一步,细菌宿主天然地吸引抗原呈递细胞(APC)的识别和细胞摄取。此外,根据细菌载体的不同,可以利用分子生物学工具来影响和/或克服有效抗原呈递的其他步骤和障碍。在这项工作中,以大肠杆菌作为细菌转染载体进行了分子工程改造。具体而言,设计了噬菌体ΦX174裂解E(LyE)基因,使其在含有不同水平溶血素O(LLO)的菌株中实现可变表达。目的是生成一种具有改善的减毒和递送特性的细菌载体。所得菌株表现出增强的基因和蛋白质释放以及可诱导的细胞死亡。此外,新载体对RAW264.7巨噬细胞APC显示出改善的基因递送和细胞毒性特征。