Structural and immunostimulatory properties of Y-shaped DNA consisting of phosphodiester and phosphorothioate oligodeoxynucleotides

JOURNAL OF CONTROLLED RELEASE

卷:148

Article

Matsuoka, Nao¹  Nishikawa, Makiya¹  Mohri, Kohta¹  Rattanakiat, Sakulrat¹  Takakura, Yoshinobu¹ 

[1] Kyoto Univ, Grad Sch Pharmaceut Sci, Dept Biopharmaceut & Drug Metab, Sakyo Ku, Kyoto 6068501, Japan

关键词: Immunostimulatory DNA;    Phosphorothioate modification;    Nano-assembly;    Thermostability;    Macrophage;    CpG motif;   

DOI  :  10.1016/j.jconrel.2010.09.019

来源: Elsevier

【 摘 要 】

V-shape formation increased the immunostimulatory activity of phosphodiester (PO) oligodeoxynucleotides (ODNs) containing CpG motif. In this study, PO CpG ODN or CpG ODN containing nuclease-resistant phosphorothioate (PS) linkages, i.e., PS CpG ODN or PO CpG ODN with three PS linkages at the both ends (PS3), was mixed with two PO- or PS ODNs to prepare Y-shaped DNA (Y-DNA) containing a potent CpG motif. The melting temperature of Y-DNA decreased with increasing number of PS linkages. Y(PS/PO/PO), which contained PS CpG ODN, showed the greatest activity to induce tumor necrosis factor-alpha release from macrophage-like RAW264.7 cells, followed by Y(PS3/PO/PO). However, the high activity of Y(PS/PO/PO) was due to that of PS CpG ODN, and Y-shape formation had no significant effect on the activity. Furthermore, PS CpG ODN of Y(PS/PO/PO) was efficiently taken up by cells, but other PO ODNs in the Y-DNA were not, indicating that PS CpG ODN in Y-DNA behave like single stranded PS CpG ODN. In quite contrast, the immunostimulatory activity of PS3 CpG ODN was significantly increased by Y-shape formation. In conclusion, Y-shape formation and PS substitution can be used simultaneously to increase the immunostimulatory activity of CpG ODN, but extensive substitution should be avoided because it diminishes the benefits of Y-shape formation. (C) 2010 Elsevier B.V. All rights reserved.

【 授权许可】

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