Design and delivery of nucleic acid drugs for optimization of DNA vaccination

用于优化 DNA 疫苗接种的核酸药物的设计和递送

• 批准号: 17390041
• 负责人: TAKAKURA Yoshinobu
• 金额: $ 9.6万
• 依托单位: Kyoto University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2005
• 资助国家: 日本
• 起止时间: 2005 至 2006
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-17390041/
• 关键词: DNA vaccination; Dendritic cell; Plasmid DNA; Delivery; Cytotoxic T cell; Intracellular trafficking; CpG motif; Hsp70; プラスミド; 抗原提示細胞; MHC class I; エレクトロポレーション

DNA vaccination, which can induce humoral immunity as well as cellular immunity, is a promising approach for treatment of various infectious diseases and tumors. However, its optimization in terms of design and delivery is required to improve the efficacy of this strategy. In the present study, we examined cellular activation of dendritic cells, the most important cells affecting the efficacy of DNA vaccination, by plasmid DNA in naked and liposomal complex form. We found that dendritic cell activation takes place depending on CpG motifs contained in DNA in both naked and complex forms, which was quite contrast to the observation in macrophages in our previous studies. We also designed novel plasmid constructs encoding antigenic peptide-heat shock protein 70 fusion proteins for controlled intracellular trafficking and subsequent improved activity in cytotoxic T cell activation. We found that intradermal injection of the constructs in combination with electroporation could effectively induce antigen specific cytotoxic T cells and antitumor effect in mice. These findings would provide the useful basis for optimization of delivery and design of plasmid DNA for DNA vaccination.

DNA疫苗接种可以诱导体液免疫和细胞免疫，是治疗各种感染性疾病和肿瘤的一种很有前途的方法。然而，为了提高该策略的有效性，需要在设计和交付方面进行优化。在本研究中，我们通过裸质粒DNA和脂质体复合物形式检测了影响DNA疫苗接种效果的最重要细胞树突状细胞的细胞活化。我们发现树突状细胞的激活依赖于DNA中包含的CpG基序，无论是裸形式还是复杂形式，这与我们之前在巨噬细胞中的观察结果形成鲜明对比。我们还设计了一种新的质粒结构，编码抗原肽-热休克蛋白70融合蛋白，用于控制细胞内运输和随后提高细胞毒性T细胞活化的活性。我们发现皮内注射这些构建物联合电穿孔可以有效地诱导抗原特异性细胞毒性T细胞，并具有抗肿瘤作用。这些发现将为DNA疫苗的传递和质粒设计的优化提供有益的依据。

项目成果

DNA and its cationic lipid complexes induce CpG motif-dependent activation of murine dendritic cells.

DNA 及其阳离子脂质复合物诱导小鼠树突状细胞的 CpG 基序依赖性激活。

• 发表时间: 2007
• 期刊: Immunology 120
• 作者: Akutsu-Yamauchi H;Hitomi J;Satoh Y;Takaharu Yoshinaga et al.
• 通讯作者: Takaharu Yoshinaga et al.

Histogram analysis of pharmacokinetic parameters by bootstrap resampling from one-point data in animal experiments.

通过动物实验中单点数据的引导重采样对药代动力学参数进行直方图分析。

• 发表时间: 2006
• 期刊: Drug Metabolism and Pharmacokinetics 21(6)
• 作者: Park;H.-R.;Chijiwa;S.;Furihata;K.;Hayakawa;Y.;Shin-ya;K.;Takemoto S et al.
• 通讯作者: Takemoto S et al.

Cellular uptake and activation characteristics of naked plasmid DNA and its cationic liposome complex in human macrophages.

• DOI: 10.1016/j.ijpharm.2005.08.020
• 发表时间: 2005-11
• 期刊: International journal of pharmaceutics
• 影响因子: 5.8
• 作者: Ikuko Yamane;M. Nishikawa;Y. Takakura

The uptake and degradation of DNA is impaired in macrophages and dendritic cells from NZB/W F(1)mice.

NZB/W F(1) 小鼠的巨噬细胞和树突状细胞中 DNA 的摄取和降解受到损害。

• 发表时间: 2005
• 期刊: Immunology Letters 101(1)
• 作者: Nakamura Y;Oosaka M;Saino T;Satoh Y:;Ogawa Y et al.
• 通讯作者: Ogawa Y et al.

Plasmid DNA uptake and subsequent cellular activation characteristics in human monocyte-derived cells in primary culture

• DOI: 10.1002/jps.20816
• 发表时间: 2007-06-01
• 期刊: JOURNAL OF PHARMACEUTICAL SCIENCES
• 影响因子: 3.8
• 作者: Fukuhara, Yuga;Naoi, Tomoyuki;Takakura, Yoshinobu
• 通讯作者: Takakura, Yoshinobu