Yildiz, S.Alpdundar, E.Gungor, B.Kahraman, T.Bayyurt, B.Gursel, I.Gursel, M.2016-02-082016-02-0820150014-2980http://hdl.handle.net/11693/24013Recognition of pathogen-derived nucleic acids by immune cells is critical for the activation of protective innate immune responses. Bacterial cyclic dinucleotides (CDNs) are small nucleic acids that are directly recognized by the cytosolic DNA sensor STING (stimulator of IFN genes), initiating a response characterized by proinflammatory cytokine and type I IFN production. Strategies to improve the immune stimulatory activities of CDNs can further their potential for clinical development. Here, we demonstrate that a simple complex of cylic-di-GMP with a cell-penetrating peptide enhances both cellular delivery and biological activity of the cyclic-di-GMP in murine splenocytes. Furthermore, our findings establish that activation of the TLR-dependent and TLR-independent DNA recognition pathways through combined use of CpG oligonucleotide (ODN) and CDN results in synergistic activity, augmenting cytokine production (IFN-α/β, IL-6, TNF-α, IP-10), costimulatory molecule upregulation (MHC class II, CD86), and antigen-specific humoral and cellular immunity. Results presented herein indicate that 3′3′-cGAMP, a recently identified bacterial CDN, is a superior stimulator of IFN genes ligand than cyclic-di-GMP in human PBMCs. Collectively, these findings suggest that the immune-stimulatory properties of CDNs can be augmented through peptide complexation or synergistic use with CpG oligonucleotide and may be of interest for the development of CDN-based immunotherapeutic agents.EnglishArginine peptide (nona-arginine)CGAMPCpG ODNCyclic-di-GMPImmunostimulationAlpha interferonArginineBacterial proteinBeta interferonCD11b antigenCD14 antigenCD45 antigenCD86 antigenCell penetrating peptideCpG oligodeoxynucleotideCyclic diguanosine monophosphateDinucleotideDNAFluorescent dyeGamma interferon inducible protein 10Glycoprotein p 15095Immunostimulating agentInterferonInterleukin 12Interleukin 6LipofectamineMajor histocompatibility antigen class 2OvalbuminToll like receptor 9Trypan blueTumor necrosis factor alphaUnclassified drugBis(3',5')-cyclic diguanylic acidCell penetrating peptideCPG-oligonucleotideCyclic GMPCyclic guanosine monophosphate-adenosine monophosphateCyclic nucleotideCytokineImmunological adjuvantInterferonMembrane proteinMPYS protein, mouseOligodeoxyribonucleotidePeptideAnimal cellAnimal experimentAnimal modelAntigen presenting cellArticleCell maturationCellular immunityComplex formationControlled studyCytokine productionCytokine releaseDrug potentiationHumanHuman cellImmunostimulationInnate immunityInternalizationLow drug doseMouseNonhumanNormal humanPeripheral blood mononuclear cellPriority journalSpleen cellTumor volumeUpregulationAnalogs and derivativesAnimalBiosynthesisC57BL mouseChemistryCpG islandCytologyDrug effectsSpleenTumor cell cultureAdjuvants, ImmunologicAnimalsCell-Penetrating PeptidesCpG IslandsCyclic GMPCytokinesHumansImmunity, InnateInterferon Type IMembrane ProteinsMiceMice, Inbred C57BLNucleotides, CyclicOligodeoxyribonucleotidesPeptidesSpleenTumor Cells, CulturedArticle10.1002/eji.201445133