Warfield Kelly L, Posten Nichole A, Swenson Dana L, Olinger Gene G, Esposito Dominic, Gillette William K, Hopkins Ralph F, Costantino Julie, Panchal Rekha G, Hartley James L, Aman M Javad, Bavari Sina
US Army Medical Research Institute of Infectious Diseases, Fort Detrick, MD 21702, USA.
J Infect Dis. 2007 Nov 15;196 Suppl 2:S421-9. doi: 10.1086/520612.
Virus-like particles (VLPs) of Ebola virus (EBOV) and Marburg virus (MARV) produced in human 293T embryonic kidney cells have been shown to be effective vaccines against filoviral infection. In this study, we explored alternative strategies for production of filovirus-like particle-based vaccines, to accelerate the development process. The goal of this work was to increase the yield of VLPs, while retaining their immunogenic properties.
Ebola and Marburg VLPs (eVLPs and mVLPs, respectively) were generated by use of recombinant baculovirus constructs expressing glycoprotein, VP40 matrix protein, and nucleoprotein from coinfected insect cells. The baculovirus-derived eVLPs and mVLPs were characterized biochemically, and then the immune responses produced by the eVLPs in insect cells were studied further.
The baculovirus-derived eVLPs elicited maturation of human myeloid dendritic cells (DCs), indicating their immunogenic properties. Mice vaccinated with insect cell-derived eVLPs generated antibody and cellular responses equivalent to those vaccinated with mammalian 293T cell-derived eVLPs and were protected from EBOV challenge in a dose-dependent manner.
Together, these data suggest that filovirus-like particles produced by baculovirus expression systems, which are amenable to large-scale production, are highly immunogenic and are suitable as safe and effective vaccines for the prevention of filoviral infection.
在人293T胚胎肾细胞中产生的埃博拉病毒(EBOV)和马尔堡病毒(MARV)的病毒样颗粒(VLPs)已被证明是预防丝状病毒感染的有效疫苗。在本研究中,我们探索了生产基于丝状病毒样颗粒疫苗的替代策略,以加速开发进程。这项工作的目标是提高VLPs的产量,同时保留其免疫原性。
通过使用表达糖蛋白、VP40基质蛋白和核蛋白的重组杆状病毒构建体感染昆虫细胞来产生埃博拉和马尔堡VLPs(分别为eVLPs和mVLPs)。对杆状病毒衍生的eVLPs和mVLPs进行生化表征,然后进一步研究昆虫细胞中eVLPs产生的免疫反应。
杆状病毒衍生的eVLPs诱导人髓样树突状细胞(DCs)成熟,表明其具有免疫原性。用昆虫细胞衍生的eVLPs接种的小鼠产生的抗体和细胞反应与用哺乳动物293T细胞衍生的eVLPs接种的小鼠相当,并以剂量依赖的方式免受EBOV攻击。
总之,这些数据表明,杆状病毒表达系统产生的丝状病毒样颗粒易于大规模生产,具有高度免疫原性,适合作为预防丝状病毒感染的安全有效疫苗。