title: Development of Nonaggregating Poly-A Tailed Immunostimulatory A/D Type CpG Oligodeoxynucleotides Applicable for Clinical Use.
abstract: Immunostimulatory CpG ODNs have been developed and utilized as TLR9-dependent innate immune activators and vaccine adjuvants. Four different types of immunostimulatory CpG ODNs (A/D, B/K, C, and P type) have been reported. A/D type ODNs are characterized by high IFN-α production but intrinsically form aggregates, hindering its good manufacturing practice grade preparation. In this study, we developed several D35-derived ODNs (a commonly used A/D type ODN), which were modified with the addition of a phosphorothioate polynucleotide tail (such as dAs40), and examined their physical properties, solubility in saline, immunostimulatory activity on human PBMCs, and vaccine adjuvant potential in monkeys. We found that two modified ODNs including D35-dAs40 and D35core-dAs40 were immunostimulatory, similar to original D35 in human PBMCs, resulting in high IFN-α secretion in a dose-dependent manner. Physical property analysis by dynamic light scattering revealed that both D35-dAs40 and D35core-dAs40 did not form aggregates in saline, which is currently impossible for the original D35. Furthermore, D35-dAs40 and D35core-dAs40 worked as better vaccine adjuvant in monkeys. These results suggested that D35-dAs40 and D35core-dAs40 are two promising prototypes of nonaggregating A/D type ODN with advantages of ease of drug preparation for clinical applications as vaccine adjuvants or IFN-α inducing immunomodifiers.
authors.affiliation: Laboratory of Adjuvant Innovation, National Institute of Biomedical Innovation, Health and Nutrition, Ibaraki, Osaka 567-0085, Japan ; Laboratory of Vaccine Science, Immunology Frontier Research Center (iFReC), Osaka University, Suita, Osaka 565-0871, Japan ; Vaccine Dynamics Project, BIKEN Innovative Vaccine Research Alliance Laboratories, Research Institute for Microbial Diseases (RIMD), Osaka University, Suita, Osaka 565-0871, Japan.
authors.forename: Taiki
authors.initials: T
authors.lastname: Aoshi
authors.affiliation: Laboratory of Adjuvant Innovation, National Institute of Biomedical Innovation, Health and Nutrition, Ibaraki, Osaka 567-0085, Japan ; Vaccine Dynamics Project, BIKEN Innovative Vaccine Research Alliance Laboratories, Research Institute for Microbial Diseases (RIMD), Osaka University, Suita, Osaka 565-0871, Japan.
authors.forename: Yasunari
authors.initials: Y
authors.lastname: Haseda
authors.affiliation: Laboratory of Adjuvant Innovation, National Institute of Biomedical Innovation, Health and Nutrition, Ibaraki, Osaka 567-0085, Japan ; Laboratory of Vaccine Science, Immunology Frontier Research Center (iFReC), Osaka University, Suita, Osaka 565-0871, Japan.
authors.forename: Kouji
authors.initials: K
authors.lastname: Kobiyama
authors.affiliation: Laboratory of Supramolecular Crystallography, Institute for Protein Research, Osaka University, Suita, Osaka 565-0871, Japan.
authors.forename: Hirotaka
authors.initials: H
authors.lastname: Narita
authors.affiliation: GeneDesign Inc., Ibaraki, Osaka 567-0085, Japan.
authors.forename: Hideaki
authors.initials: H
authors.lastname: Sato
authors.affiliation: GeneDesign Inc., Ibaraki, Osaka 567-0085, Japan.
authors.forename: Hirokazu
authors.initials: H
authors.lastname: Nankai
authors.affiliation: Department of Chemistry and Biochemistry, University of Kitakyushu, Kitakyushu, Fukuoka 808-0135, Japan.
authors.forename: Shinichi
authors.initials: S
authors.lastname: Mochizuki
authors.affiliation: Department of Chemistry and Biochemistry, University of Kitakyushu, Kitakyushu, Fukuoka 808-0135, Japan.
authors.forename: Kazuo
authors.initials: K
authors.lastname: Sakurai
authors.affiliation: Corporation for Production and Research of Laboratory Primates, Tsukuba, Ibaraki 305-0843, Japan.
authors.forename: Yuko
authors.initials: Y
authors.lastname: Katakai
authors.affiliation: Tsukuba Primate Research Center, National Institute of Biomedical Innovation, Tsukuba, Ibaraki 305-0843, Japan.
authors.forename: Yasuhiro
authors.initials: Y
authors.lastname: Yasutomi
authors.affiliation: Laboratory of Vaccine Science, Immunology Frontier Research Center (iFReC), Osaka University, Suita, Osaka 565-0871, Japan.
authors.forename: Etsushi
authors.initials: E
authors.lastname: Kuroda
authors.affiliation: Laboratory of Malaria Immunology, Immunology Frontier Research Center (iFReC), Osaka University, Suita, Osaka 565-0871, Japan.
authors.forename: Cevayir
authors.initials: C
authors.lastname: Coban
authors.affiliation: Laboratory of Adjuvant Innovation, National Institute of Biomedical Innovation, Health and Nutrition, Ibaraki, Osaka 567-0085, Japan ; Laboratory of Vaccine Science, Immunology Frontier Research Center (iFReC), Osaka University, Suita, Osaka 565-0871, Japan.
authors.forename: Ken J
authors.initials: KJ
authors.lastname: Ishii
mesh_terms: D000276:Adjuvants, Immunologic; D000818:Animals; D016207:Cytokines / Q000096:biosynthesis; D004305:Dose-Response Relationship, Drug; D004306:Dose-Response Relationship, Immunologic; D005229:Fatty Acids, Monounsaturated / Q000737:chemistry; D006801:Humans; D007252:Influenza Vaccines / Q000276:immunology; D007963:Leukocytes, Mononuclear / Q000187:drug effects / Q000276:immunology / Q000378:metabolism; D008252:Macaca fascicularis; D009838:Oligodeoxyribonucleotides / Q000008:administration & dosage / Q000737:chemistry / Q000276:immunology*; D011061:Poly A / Q000737:chemistry; D000644:Quaternary Ammonium Compounds / Q000737:chemistry; D012995:Solubility
chemical_list: D000276:Adjuvants, Immunologic; C408982:CPG-oligonucleotide; D016207:Cytokines; D005229:Fatty Acids, Monounsaturated; D007252:Influenza Vaccines; D009838:Oligodeoxyribonucleotides; D000644:Quaternary Ammonium Compounds; D011061:Poly A; C070046:1,2-dioleoyloxy-3-(trimethylammonium)propane