title: Potential immunological adjuvant of `K'-type CpG-oligodeoxynucleotides enhanced the cell proliferation and IL-6 mRNA transcription in canine B cells.
abstract: CpG oligodeoxynucleotides (CpG-ODNs) are ligands for toll-like receptor 9 (TLR9), signaling of which plays a role in innate immunity by inducing T helper 1 (TH1)-cell responses and pro-inflammatory cytokine production. The activation of TLR9 signaling is considered to be effective for the therapy of cancer, infectious diseases, and allergies and preclinical studies using CpG-ODNs have been performed in dogs and humans. In order to investigate the precise mechanisms responsible for the effect of CpG-ODNs in dogs, we examined their role in cell proliferation and cytokine gene expression in canine B cells. Canine B cells were collected by a magnetic cell isolation method using anti-CD21 antibody. Flow cytometric analysis for the intracellular CD79α revealed the purity of canine B cells to be as high as 90.2 ± 2.1%. Transcription of TLR2, TLR4, and TLR9 mRNA on canine CD21(+) cells was confirmed by reverse-transcript polymerase chain reaction (RT-PCR). CpG-ODNs induced dose-dependent proliferation of canine CD21(+) cells (P<0.05 compared with control-ODNs) detected by BrdU incorporation. Quantification of IL-6, IL-10, and IL-12p40 mRNA transcription on canine CD21(+) cells revealed that CpG-ODNs enhanced IL-6 mRNA transcription but not IL-10 and IL-12p40 mRNA transcription (P<0.05 compared with control-ODNs). These responses to CpG-ODNs in the canine B cells indicated that CpG-ODNs would be useful as an immunological adjuvant for vaccine in dogs.
authors.affiliation: Department of Veterinary Microbiology, Azabu University, Kanagawa, Japan.
authors.forename: Shinpei
authors.initials: S
authors.lastname: Kawarai
authors.forename: Kaoru
authors.initials: K
authors.lastname: Sato
authors.forename: Anna
authors.initials: A
authors.lastname: Horiguchi
authors.forename: Keigo
authors.initials: K
authors.lastname: Kurata
authors.forename: Akio
authors.initials: A
authors.lastname: Kiuchi
authors.forename: Hajime
authors.initials: H
authors.lastname: Tsujimoto
authors.forename: Masahiro
authors.initials: M
authors.lastname: Sakaguchi
mesh_terms: D000276:Adjuvants, Immunologic / Q000494:pharmacology*; D000818:Animals; D001402:B-Lymphocytes / Q000276:immunology*; D048708:Cell Growth Processes / Q000187:drug effects; D004285:Dogs / Q000276:immunology*; D005260:Female; D005434:Flow Cytometry; D007378:Interleukins / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology; D008297:Male; D009838:Oligodeoxyribonucleotides / Q000494:pharmacology*; D012313:RNA / Q000235:genetics / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000662:veterinary; D018709:Statistics, Nonparametric; D051193:Toll-Like Receptors / Q000235:genetics / Q000276:immunology
chemical_list: D000276:Adjuvants, Immunologic; C408982:CPG-oligonucleotide; D007378:Interleukins; D009838:Oligodeoxyribonucleotides; D051193:Toll-Like Receptors; D012313:RNA