Enhanced immunostimulatory activity of oligodeoxynucleotides by Y-shape formation.
DNA containing unmethylated CpG dinucleotides (CpG DNA) is a potent activator of innate and acquired immune responses. Although the sequence-specific immunostimulatory activity of CpG DNA has been extensively explored, little information is available about the importance of the stereochemical properties of CpG DNA. In this study, Y-shaped oligodeoxynucleotides (Y-ODNs) were prepared using three ODNs with the halves of each ODN being partially complementary to a half of the other two ODNs. Y-ODN induced greater amounts of tumour necrosis factor-alpha and interleukin-6 from RAW264.7 macrophage-like cells than did conventional single-stranded ODN (ssODN) or double-stranded ODN (dsODN). The Y-ODN was less stable in serum than dsODN, but greater amounts of Y-ODN were taken up by macrophage-like cells compared with dsODN. A newly designed Y-ODN containing three potent CpG motifs generated significantly higher levels of cytokines compared with dsODN containing the identical sequences. These results indicate that the Y-shaped form of ODN is a novel, reproducible and reliable approach to enhancing the immunostimulatory activity of ODNs.
Nishikawa
Makiya
M
Department of Biopharmaceutics and Drug Metabolism, Graduate School of Pharmaceutical Sciences, Kyoto University, Sakyo-ku, Kyoto, Japan.
Matono
Mitsuhiro
M
Rattanakiat
Sakulrat
S
Matsuoka
Nao
N
Takakura
Yoshinobu
Y
D000276:Adjuvants, Immunologic / Q000737:chemistry*; D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D055598:Chemical Phenomena; D002627:Chemistry, Physical; D018899:CpG Islands / Q000276:immunology; D017077:Culture Media, Conditioned; D016207:Cytokines / Q000378:metabolism; D008264:Macrophages / Q000276:immunology / Q000378:metabolism; D051379:Mice; D008969:Molecular Sequence Data; D009838:Oligodeoxyribonucleotides / Q000737:chemistry / Q000276:immunology* / Q000493:pharmacokinetics; D013329:Structure-Activity Relationship; D051217:Toll-Like Receptor 9 / Q000378:metabolism
D000276:Adjuvants, Immunologic; D017077:Culture Media, Conditioned; D016207:Cytokines; D009838:Oligodeoxyribonucleotides; C495370:Tlr9 protein, mouse; D051217:Toll-Like Receptor 9