Department of Biopharmaceutics and Drug Metabolism, Graduate School of Pharmaceutical Sciences, Kyoto University, Sakyo-ku, Kyoto, Japan.
Department of Biopharmaceutics and Drug Metabolism, Graduate School of Pharmaceutical Sciences, Kyoto University, Sakyo-ku, Kyoto, Japan; Institute for Integrated Cell-Material Sciences (iCeMS), Kyoto University, Sakyo-ku, Kyoto, Japan.
Nanomedicine. 2014 May;10(4):765-74. doi: 10.1016/j.nano.2013.11.017. Epub 2013 Dec 10.
Investigation of mouse macrophage-like RAW264.7 cells showed that the immunostimulatory activity of CpG DNA is increased by formation of polypod-like structured DNA (polypodna), an assembly consisting of three or more oligodeoxynucleotides. To apply CpG polypodna to immunotherapy, its activity was examined in murine dendritic DC2.4 cells, splenic macrophages, and bone marrow-derived dendritic cells (BMDCs). In all cell types, increasing the pod number increased the cellular uptake of DNA and cytokine release. No significant release of cytokines was observed in macrophages lacking Toll-like receptor 9. Similar results were obtained after intradermal injection of polypodna. The polypodna preparations produced significantly higher amounts of interferon α in human peripheral blood mononuclear cells (PBMCs) compared with single-stranded DNA. The conditioned medium of hexapodna-treated human PBMCs effectively inhibited the activity of a hepatitis C virus subgenomic replicon reporter system. These results indicate that polypodna preparations are useful as an immunostimulator.
This study demonstrates the utility of polypoid-like structured DNA (polypodna) preparations as potent immunostimulators in a murine model.
对小鼠巨噬细胞样 RAW264.7 细胞的研究表明，CpG DNA 的免疫刺激活性通过形成多足样结构 DNA（多足 DNA）而增加，该多足结构由三个或更多的寡脱氧核苷酸组成。为了将 CpG 多足 DNA 应用于免疫治疗，研究了其在小鼠树突状 DC2.4 细胞、脾巨噬细胞和骨髓来源的树突状细胞（BMDC）中的活性。在所有细胞类型中，增加足的数量都会增加细胞对 DNA 的摄取和细胞因子的释放。在缺乏 Toll 样受体 9 的巨噬细胞中未观察到细胞因子的显著释放。在皮内注射多足 DNA 后也获得了类似的结果。与单链 DNA 相比，多足 DNA 制剂在人外周血单核细胞（PBMC）中产生了明显更高量的干扰素-α。用六足 DNA 处理的人 PBMC 的条件培养基可有效抑制丙型肝炎病毒亚基因组复制子报告系统的活性。这些结果表明，多足 DNA 制剂可用作有效的免疫刺激剂。
本研究证明了多足样结构 DNA（多足 DNA）制剂在小鼠模型中作为有效免疫刺激剂的用途。
