Suitable Promoter for DNA Vaccination Using a pDNA Ternary Complex

Pharmaceutics Pub Date : 2024-05-17 DOI:10.3390/pharmaceutics16050679

Tomoaki Kurosaki, Hiroki Nakamura, Hitoshi Sasaki, Yukinobu Kodama

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Abstract

In this study, we evaluated the effect of several promoters on the transfection activity and immune-induction efficiency of a plasmid DNA (pDNA)/polyethylenimine/γ-polyglutamic acid complex (pDNA ternary complex). Model pDNAs encoding firefly luciferase (Luc) were constructed with several promoters, such as simian virus 40 (SV40), eukaryotic elongation factor 1 alpha (EF1), cytomegalovirus (CMV), and chicken beta actin hybrid (CBh) (pSV40-Luc, pEF1-Luc, pCMV-Luc, and pCBh-Luc, respectively). Four types of pDNA ternary complexes, each with approximately 145-nm particle size and −30-mV ζ-potential, were stably constructed. The pDNA ternary complex containing pSV40-Luc showed low gene expression, but the other complexes containing pEF1-Luc, pCMV-Luc, and pCBh-Luc showed high gene expression in DC2.4 cells and spleen after intravenous administration. After immunization using various pDNA encoding ovalbumin (OVA) such as pEF1-OVA, pCMV-OVA, and pCBh-OVA, only the pDNA ternary complex containing pCBh-OVA showed significant anti-OVA immunoglobulin G (IgG) induction. In conclusion, our results showed that the CBh promoter is potentially suitable for use in pDNA ternary complex-based DNA vaccination.

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使用 pDNA 三元复合物接种 DNA 疫苗的合适启动子

本研究评估了几种启动子对质粒 DNA（pDNA）/聚乙烯亚胺/γ-聚谷氨酸复合物（pDNA 三元复合物）转染活性和免疫诱导效率的影响。用几种启动子构建了编码萤火虫荧光素酶（Luc）的模型 pDNA（分别为 pSV40-Luc、pEF1-Luc、pCMV-Luc 和 pCBh-Luc），这些启动子包括猿猴病毒 40（SV40）、真核延伸因子 1 alpha（EF1）、巨细胞病毒（CMV）和鸡 beta 肌动蛋白杂交体（CBh）。我们稳定地构建了四种 pDNA 三元复合物，每种复合物的粒径约为 145 纳米，ζ电位为-30 毫伏。含有 pSV40-Luc 的 pDNA 三元复合物的基因表达量较低，而含有 pEF1-Luc、pCMV-Luc 和 pCBh-Luc 的其他复合物在静脉注射后在 DC2.4 细胞和脾脏中的基因表达量较高。使用 pEF1-OVA、pCMV-OVA 和 pCBh-OVA 等各种编码卵清蛋白（OVA）的 pDNA 进行免疫后，只有含有 pCBh-OVA 的 pDNA 三元复合物显示出显著的抗 OVA 免疫球蛋白 G（IgG）诱导作用。总之，我们的研究结果表明，CBh 启动子可能适用于基于 pDNA 三元复合物的 DNA 疫苗接种。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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Pharmaceutics

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