title: Loxoribine, a selective Toll-like receptor 7 agonist, induces maturation of human monocyte-derived dendritic cells and stimulates their Th-1- and Th-17-polarizing capability.
abstract: Recently, a guanosine analog, 7-allyl-7,8-dihydro-8-oxo-guanosine (loxoribine), has been identified as a selective Toll-like receptor (TLR)7 agonist. Bearing in mind the controversy regarding the expression of TLR7 by human myeloid dendritic cells (DCs) and its significance for functions of these cells, the goal of this study was to investigate the effect of loxoribine on differentiation, maturation and functions of human monocyte-derived (Mo)DCs. Immature MoDCs were obtained by cultivation of monocytes for 6 days with recombinant granulocyte macrophage-colony stimulating factor (GM-CSF) and interleukin (IL)-4. These cells were stimulated with loxoribine (250 μM) for an additional 48 h. Phenotypic properties of MoDCs were determined by flow cytometry, cytokine production was assayed by ELISA, whereas their allostimulatory capability was tested using a mixed leukocyte reaction. We showed that loxoribine up-regulated the expression of TLR7, CD40, CD54, CD80, CD83 and CCR7 and stimulated the production of IL-12, IL-23, IL-27 and IL-10 by MoDCs, whereas the level of interferon (IFN)-β was not modulated. Allogeneic CD4(+)T cells in co-culture with loxoribine-treated MoDCs proliferated more strongly, at lower DC/CD4(+)T-cell ratio (1:80), and secreted significantly higher levels of IL-17 and IFN-γ compared to the cultures with control MoDCs. The stimulatory effect of loxoribine on T helper (Th)1 polarization capability of MoDCs was further potentiated by ligation of CD40. In conclusion, our results show that loxoribine stimulated differentiation, maturation, allostimulatory as well as Th1 and Th17 polarization capability of human MoDCs and suggests that these effects might be associated with up-regulation of TLR7 expression, but not increased IFN-β production.
authors.affiliation: Institute for Medical Research, Military Medical Academy, Belgrade, Serbia.
authors.forename: Tanja
authors.initials: T
authors.lastname: Dzopalic
authors.forename: Ana
authors.initials: A
authors.lastname: Dragicevic
authors.forename: Sasa
authors.initials: S
authors.lastname: Vasilijic
authors.forename: Dragana
authors.initials: D
authors.lastname: Vucevic
authors.forename: Ivana
authors.initials: I
authors.lastname: Majstorovic
authors.forename: Biljana
authors.initials: B
authors.lastname: Bozic
authors.forename: Bela
authors.initials: B
authors.lastname: Balint
authors.forename: Miodrag
authors.initials: M
authors.lastname: Colic
mesh_terms: D015703:Antigens, CD / Q000032:analysis / Q000276:immunology; D018122:B7-1 Antigen / Q000032:analysis / Q000276:immunology; D015496:CD4-Positive T-Lymphocytes / Q000187:drug effects / Q000276:immunology; D019013:CD40 Antigens / Q000032:analysis / Q000276:immunology; D002454:Cell Differentiation / Q000187:drug effects / Q000276:immunology; D002478:Cells, Cultured; D018920:Coculture Techniques; D003713:Dendritic Cells / Q000187:drug effects* / Q000276:immunology; D016178:Granulocyte-Macrophage Colony-Stimulating Factor / Q000494:pharmacology; D006151:Guanosine / Q000031:analogs & derivatives* / Q000494:pharmacology; D006801:Humans; D007136:Immunoglobulins / Q000032:analysis / Q000276:immunology; D018799:Intercellular Adhesion Molecule-1 / Q000032:analysis / Q000276:immunology; D007371:Interferon-gamma / Q000032:analysis / Q000276:immunology / Q000378:metabolism; D015847:Interleukin-4 / Q000276:immunology / Q000494:pharmacology; D007378:Interleukins / Q000032:analysis / Q000096:biosynthesis / Q000276:immunology; D007963:Leukocytes, Mononuclear / Q000276:immunology / Q000378:metabolism; D008562:Membrane Glycoproteins / Q000032:analysis / Q000276:immunology; D054400:Receptors, CCR7 / Q000032:analysis / Q000276:immunology; D011994:Recombinant Proteins; D018417:Th1 Cells / Q000187:drug effects* / Q000276:immunology; D058504:Th17 Cells / Q000187:drug effects* / Q000276:immunology; D051199:Toll-Like Receptor 7 / Q000819:agonists* / Q000032:analysis / Q000276:immunology; D015854:Up-Regulation; D000097802:CD83 Antigen
chemical_list: D015703:Antigens, CD; D018122:B7-1 Antigen; D019013:CD40 Antigens; D016178:Granulocyte-Macrophage Colony-Stimulating Factor; D006151:Guanosine; D007136:Immunoglobulins; D018799:Intercellular Adhesion Molecule-1; D007371:Interferon-gamma; D015847:Interleukin-4; D007378:Interleukins; D008562:Membrane Glycoproteins; D054400:Receptors, CCR7; D011994:Recombinant Proteins; D051199:Toll-Like Receptor 7; D000097802:CD83 Antigen; C515370:CCR7 protein, human; C068562:loxoribine