Broderick Kate E, Khan Amir S, Sardesai Niranjan Y
Inovio Pharmaceuticals, 1787 Sentry Parkway West, Blue Bell PA, 19422, USA,
Methods Mol Biol. 2014;1143:123-30. doi: 10.1007/978-1-4939-0410-5_8.
DNA vaccines are a next generation branch of vaccines which offer major benefits over their conventional counterparts. However, to be effective in large mammals and humans, an enhancing delivery technology is required. Electroporation is a physical technique which results in improved delivery of large molecules through the cell membrane. In the case of plasmid DNA, electroporation enhances both the uptake and expression of the delivered DNA. The skin is an attractive tissue for DNA vaccination in a clinical setting due to the accessibility of the target, the ease of monitoring, and most importantly the immunocompetent nature of the dermis. Electroporation in the skin has the benefit of being minimally invasive and generally well tolerated. Previous studies have determined that optimized electroporation parameters (such as electrical field intensity, pulse length, pulse width, and plasmid formulation) majorly impact the efficiency of DNA delivery to the skin. We provide an overview of DNA vaccination in skin and muscle. In addition, we detail a protocol for the successful intradermal electroporation of plasmid DNA to guinea pig skin, an excellent dermatological animal model. The work detailed here suggests that the technique is safe and effective and could be highly applicable to a clinical setting.
DNA疫苗是疫苗的新一代分支，与传统疫苗相比具有诸多主要优势。然而，要在大型哺乳动物和人类中发挥作用，需要一种增强递送技术。电穿孔是一种物理技术，可使大分子更有效地通过细胞膜递送。对于质粒DNA而言，电穿孔可增强递送DNA的摄取和表达。在临床环境中，由于靶点易于接近、易于监测，最重要的是真皮具有免疫活性，皮肤是进行DNA疫苗接种的理想组织。皮肤电穿孔具有微创且通常耐受性良好的优点。先前的研究已确定，优化的电穿孔参数（如电场强度、脉冲长度、脉冲宽度和质粒配方）对DNA递送至皮肤的效率有重大影响。我们概述了皮肤和肌肉中的DNA疫苗接种。此外，我们详细介绍了将质粒DNA成功皮内电穿孔至豚鼠皮肤的方案，豚鼠是一种优秀的皮肤病学动物模型。此处详述的工作表明该技术安全有效，且可能高度适用于临床环境。
