University of South Florida, College of Medicine, 12901 Bruce B. Downs Blvd., Tampa, FL 33612, USA; Old Dominion University, Center for Bioelectrics, 4211 Monarch Way, Suite 300, Norfolk, VA 23508, USA.
Bioelectrochemistry. 2013 Dec;94:1-6. doi: 10.1016/j.bioelechem.2013.04.004. Epub 2013 Apr 26.
Gene electrotransfer (GET) enhances delivery of DNA vaccines by increasing both gene expression and immune responses. Our lab has developed the multi-electrode array (MEA) for DNA delivery to skin. The MEA was used at constant pulse duration (150 ms) and frequency (6.67 Hz). In this study, delivery parameters including applied voltage (5-45 V), amount of plasmid (100-300 μg), and number of treatments (2-3) were evaluated for delivery of a DNA vaccine. Mice were intradermally injected with plasmid expressing Bacillus anthracis protective antigen with or without GET and αPA serum titers measured. Within this experiment no significant differences were noted in antibody levels from varying dose or treatment number. However, significant differences were measured from applied voltages of 25 and 35 V. These voltages generated antibody levels between 20,000 and 25,000. Serum from animals vaccinated with these conditions also resulted in toxin neutralization in 40-60% of animals. Visual damage was noted at MEA conditions of 40 V. No damage was noted either visually or histologically from conditions of 35 V or below. These results reflect the importance of establishing appropriate electrical parameters and the potential for the MEA in non-invasive DNA vaccination against B. anthracis.
基因电转移(GET)通过增加基因表达和免疫反应来增强 DNA 疫苗的传递。我们的实验室已经开发出用于向皮肤输送 DNA 的多电极阵列(MEA)。MEA 的脉冲持续时间(150 ms)和频率(6.67 Hz)保持不变。在这项研究中,评估了包括应用电压(5-45 V)、质粒量(100-300 μg)和处理次数(2-3)在内的输送参数,以输送 DNA 疫苗。用表达炭疽芽孢杆菌保护性抗原的质粒对小鼠进行皮内注射,并用 GET 和 αPA 血清滴度进行测量。在该实验中,从不同剂量或处理次数变化中,抗体水平没有明显差异。然而,从 25 和 35 V 的应用电压中测量到了显著差异。这些电压产生的抗体水平在 20,000 到 25,000 之间。用这些条件接种的动物的血清也导致 40-60%的动物毒素中和。在 40 V 的 MEA 条件下观察到视觉损伤。在 35 V 或以下的条件下,无论是从视觉上还是组织学上都没有观察到损伤。这些结果反映了确定适当的电参数的重要性以及 MEA 在针对炭疽芽孢杆菌的非侵入性 DNA 疫苗接种中的潜力。