title: Development of CpG-oligodeoxynucleotides for effective activation of rabbit TLR9 mediated immune responses.
abstract: CpG-oligodeoxynucleotides (CpG-ODN) are potent immune stimuli being developed for use as adjuvants in different species. Toll-like receptor 9 (TLR9) is the cellular receptor for CpG-ODN in mammalian cells. The CpG-ODN with 18-24 deoxynucleotides that are in current use for human and mouse cells, however, have low activity with rabbit TLR9. Using a cell-based activation assay, we developed a type of CpG-ODN containing a GACGTT or AACGTT motif in 12 phosphorothioate-modified deoxynucleotides with potent stimulatory activity for rabbit TLR9. The developed CpG-ODN have higher activities than other developed CpG-ODN in eliciting antigen-nonspecific immune responses in rabbit splenocytes. When mixed with an NJ85 peptide derived from rabbit hemorrhagic disease virus, they had potent activities to boost an antigen-specific T cell activation and antibody production in rabbits. Compared to Freund's adjuvant, the developed CpG-ODN are capable of boosting a potent and less toxic antibody response. The results of this study suggest that both the choice of CpG-motif and its length are important factors for CpG-ODN to effectively activate rabbit TLR9 mediated immune responses.
authors.affiliation: Immunology Research Center, National Health Research Institutes, Miaoli, Taiwan; Program in Environmental and Occupational Medicine, Kaohsiung Medical University, Kaohsiung, Taiwan.
authors.forename: Tsung-Hsien
authors.initials: TH
authors.lastname: Chuang
authors.affiliation: Immunology Research Center, National Health Research Institutes, Miaoli, Taiwan.
authors.forename: Chao-Yang
authors.initials: CY
authors.lastname: Lai
authors.affiliation: Institute of Biochemistry and Molecular Biology, National Yang-Ming University, Taipei, Taiwan.
authors.forename: Ping-Hui
authors.initials: PH
authors.lastname: Tseng
authors.affiliation: Department of Biological Science and Technology, National Chiao Tung University, Hsinchu, Taiwan.
authors.forename: Chiun-Jye
authors.initials: CJ
authors.lastname: Yuan
authors.affiliation: Institute of Molecular Medicine, National Taiwan University, Taipei, Taiwan.
authors.forename: Li-Chung
authors.initials: LC
authors.lastname: Hsu
mesh_terms: D000276:Adjuvants, Immunologic / Q000737:chemistry / Q000494:pharmacology*; D000818:Animals; D000917:Antibody Formation / Q000187:drug effects; D001483:Base Sequence; D016207:Cytokines / Q000378:metabolism; D057809:HEK293 Cells; D017929:Hemorrhagic Disease Virus, Rabbit / Q000378:metabolism; D006801:Humans; D007075:Immunoglobulin M / Q000378:metabolism; D008213:Lymphocyte Activation / Q000187:drug effects*; D009838:Oligodeoxyribonucleotides / Q000737:chemistry / Q000494:pharmacology*; D010455:Peptides / Q000737:chemistry / Q000494:pharmacology; D010710:Phosphates / Q000737:chemistry; D011817:Rabbits; D013154:Spleen / Q000187:drug effects / Q000378:metabolism; D013601:T-Lymphocytes / Q000276:immunology / Q000378:metabolism; D051217:Toll-Like Receptor 9 / Q000378:metabolism*
chemical_list: D000276:Adjuvants, Immunologic; C408982:CPG-oligonucleotide; D016207:Cytokines; D007075:Immunoglobulin M; D009838:Oligodeoxyribonucleotides; D010455:Peptides; D010710:Phosphates; D051217:Toll-Like Receptor 9; C083120:phosphorodithioic acid