US Army Medical Research Institute for Infectious Disease, Fort Detrick, MD, USA.
Arcturus Therapeutics, San Diego, CA, USA.
Sci Rep. 2020 May 29;10(1):8764. doi: 10.1038/s41598-020-65059-0.
The use of nucleic acid as a drug substance for vaccines and other gene-based medicines continues to evolve. Here, we have used a technology originally developed for mRNA in vivo delivery to enhance the immunogenicity of DNA vaccines. We demonstrate that neutralizing antibodies produced in rabbits and nonhuman primates injected with lipid nanoparticle (LNP)-formulated Andes virus or Zika virus DNA vaccines are elevated over unformulated vaccine. Using a plasmid encoding an anti-poxvirus monoclonal antibody (as a reporter of protein expression), we showed that improved immunogenicity is likely due to increased in vivo DNA delivery, resulting in more target protein. Specifically, after four days, up to 30 ng/mL of functional monoclonal antibody were detected in the serum of rabbits injected with the LNP-formulated DNA. We pragmatically applied the technology to the production of human neutralizing antibodies in a transchromosomic (Tc) bovine for use as a passive immunoprophylactic. Production of neutralizing antibody was increased by >10-fold while utilizing 10 times less DNA in the Tc bovine. This work provides a proof-of-concept that LNP formulation of DNA vaccines can be used to produce more potent active vaccines, passive countermeasures (e.g., Tc bovine), and as a means to produce more potent DNA-launched immunotherapies.
核酸作为疫苗和其他基于基因的药物的药物物质的应用不断发展。在这里,我们使用了最初为体内 mRNA 传递开发的技术来增强 DNA 疫苗的免疫原性。我们证明,用脂质纳米颗粒(LNP)配制的安第斯病毒或寨卡病毒 DNA 疫苗注射的兔子和非人类灵长类动物产生的中和抗体高于未配制的疫苗。我们使用编码抗痘病毒单克隆抗体的质粒(作为蛋白表达的报告),表明免疫原性的提高可能是由于体内 DNA 传递增加,导致更多的靶蛋白。具体来说,在用 LNP 配制的 DNA 注射的兔子的血清中,四天后可检测到高达 30ng/mL 的功能性单克隆抗体。我们务实的将该技术应用于转染色体(Tc)牛中产生人中和抗体,用作被动免疫预防。在 Tc 牛中利用的 DNA 减少了 10 倍,但产生的中和抗体增加了 10 倍以上。这项工作提供了一个概念验证,即 DNA 疫苗的 LNP 配方可用于生产更有效的主动疫苗、被动对策(例如,Tc 牛),并可用于生产更有效的 DNA 启动的免疫疗法。