Lin, S.-Y.S.-Y.LinYao, B.-Y.B.-Y.YaoHu, C.-M.J.C.-M.J.HuHUI-WEN CHEN2020-10-152020-10-152020https://scholars.lib.ntu.edu.tw/handle/123456789/517085Background: Poultry vaccine has limited choices of adjuvants and is facing severe threat of infectious diseases due to ineffective of widely used commercial vaccines. Thus, development of novel adjuvant that offers safe and effective immunity is of urgent need. Materials and Methods: The present research engineers a novel chicken adjuvant with potent immune-potentiating capability by incorporating avian toll-like receptor 21 (TLR21) agonist CpG ODN 2007 with a poly(lactic-co-glycolic acid) (PLGA)-based hollow nanopar-ticle platform (CpG-NP), which subsequently assessed ex vivo and in vivo. Results: CpG-NPs with an average diameter of 164 nm capable of sustained release of CpG for up to 96 hours were successfully prepared. With the ex vivo model of chicken bone marrow-derived dendritic cells (chBMDCs), CpG-NP was engulfed effectively and found to induce DC maturation, promoting dendrite formation and upregulation of CD40, CD80 and CCR7. In addition to enhanced expression of IL-1β, IL-6, IL-12 and IFN-γ, 53/84 immune-related genes were found to be stimulated in CpG-NP-treated chBMDCs, whereas only 39 of such genes were stimulated in free CpG-treated cells. These upregulated genes suggest immune skewing toward T helper cell 1 bias and evidence of improved mucosal immunity upon vaccination with the CpG-NP. The CpG-NP-treated chBMDCs showed protective effects to DF-1 cells against avian influenza virus H6N1 infection. Upon subsequent coupling with infectious bronchitis virus subunit antigen administration, chickens were immunostimulated to acquire higher humoral immune response and protective response against viral challenge. Conclustion: This work presents a novel hollow CpG-NP formulation, demonstrating effective and long-lasting immunostimulatory ability ex vivo and in vivo for chickens, as systemically compared to free CpG. This enhanced immune stimulation benefits from high stability and controlled release of internal component of nanoparticles that improve cellular delivery, lymphoid organ targeting and sustainable DC activation. CpG-NP has broad application potential in antiviral and vaccine development. ? 2020 Lin et al.Adjuvant; Antiviral response; CpG; Infectious bronchitis virus; Influenza virus; Polymeric hollow nanoparticle; Vaccine[SDGs]SDG3B7 antigen; CD40 antigen; chemokine receptor CCR7; CpG oligodeoxynucleotide; gamma interferon; interleukin 12; interleukin 1beta; interleukin 6; nanoparticle; polyglactin; toll like receptor agonist; virus spike protein; virus vaccine; antivirus agent; CPG-oligonucleotide; immunological adjuvant; nanoparticle; oligodeoxyribonucleotide; polymer; vaccine; animal cell; animal experiment; animal tissue; Article; Avian infectious bronchitis virus; avian influenza virus; bone marrow derived dendritic cell; cell maturation; cell protection; chicken; controlled release formulation; controlled study; dendrite; drug delivery system; drug dosage form comparison; drug safety; embryo; ex vivo study; helper cell; humoral immunity; immune response; immune-related gene; immunostimulation; in vivo study; influenza; influenza vaccination; innate immunity; lymphoid organ; mucosal immunity; nonhuman; receptor binding; UMNSAH/DF-1 cell line; upregulation; animal; cell survival; chemistry; chicken; dendritic cell; dog; drug effect; immunity; immunization; immunology; MDCK cell line; ultrastructure; Adjuvants, Immunologic; Animals; Antiviral Agents; Cell Survival; Chickens; Dendritic Cells; Dogs; Immunity; Immunity, Humoral; Immunization; Infectious bronchitis virus; Madin Darby Canine Kidney Cells; Nanoparticles; Oligodeoxyribonucleotides; Polylactic Acid-Polyglycolic Acid Copolymer; Polymers; Vaccinesjournal article10.2147/IJN.S2414922-s2.0-85085583265