title: Production of large numbers of plasmacytoid dendritic cells with functional activities from CD34(+) hematopoietic progenitor cells: use of interleukin-3.
abstract: Plasmacytoid dendritic cells (pDC), a subset of dendritic cells characterized by a rapid and massive type-I interferon secretion through the Toll-like receptor pathway in response to viral infection, play important roles in the pathogenesis of several diseases, such as chronic viral infections (e.g., hepatitis C virus, human immunodeficiency virus), autoimmunity (e.g., psoriasis, systemic lupus erythematosus), and cancer. As pDC represent a rare cell type in the peripheral blood, the goal of this study was to develop a new method to efficiently generate large numbers of cells from a limited number of CD34(+) cord blood progenitors to provide a tool to resolve important questions about how pDC mediate tolerance, autoimmunity, and cancer. Human CD34(+) hematopoietic progenitor cells isolated from cord blood were cultured with a combination of Flt3-ligand (Flt3L), thrombopoietin (TPO), and one of the following cytokine: interleukin (IL)-3, interferon-β(IFN-β), or prostaglandin E2(PGE(2)). Cells obtained in the different culture conditions were analyzed for their phenotype and functional characteristics. The addition of IL-3 cooperates with Flt3L and TPO in the induction of pDC from CD34(+) hematopoietic progenitor cells. Indeed, Flt3L/TPO alone or supplemented with prostaglandin E2 or interferon-β produced smaller amounts of pDC from hematopoietic progenitor cells. In addition, pDC generated in Flt3L/TPO/IL-3 cultures exhibited morphological, immunohistochemical, and functional features of peripheral blood pDC. We showed that IL-3, in association with Flt3L and TPO, provides an advantageous tool for large-scale generation of pDC. This culture condition generated, starting from 2 × 10(5) CD34(+) cells, up to 2.6 × 10(6) pDC presenting features of blood pDC.
authors.affiliation: Laboratory of Experimental Pathology, GIGA-Cancer (Center for Experimental Cancer Research), University of Liege, Liege, Belgium. stephanie.demoulin@ulg.ac.be
authors.forename: Stéphanie
authors.initials: S
authors.lastname: Demoulin
authors.forename: Patrick
authors.initials: P
authors.lastname: Roncarati
authors.forename: Philippe
authors.initials: P
authors.lastname: Delvenne
authors.forename: Pascale
authors.initials: P
authors.lastname: Hubert
mesh_terms: D000328:Adult; D018929:Cell Culture Techniques; D002454:Cell Differentiation / Q000187:drug effects; D002478:Cells, Cultured / Q000166:cytology / Q000187:drug effects; D003470:Culture Media / Q000494:pharmacology; D003713:Dendritic Cells / Q000166:cytology* / Q000276:immunology; D015232:Dinoprostone / Q000494:pharmacology; D004357:Drug Synergism; D005312:Fetal Blood / Q000166:cytology; D006412:Hematopoietic Stem Cells / Q000166:cytology / Q000187:drug effects*; D006801:Humans; D016130:Immunophenotyping; D007231:Infant, Newborn; D016899:Interferon-beta / Q000494:pharmacology; D007377:Interleukin-3 / Q000494:pharmacology*; D007963:Leukocytes, Mononuclear / Q000166:cytology / Q000187:drug effects; D008565:Membrane Proteins / Q000494:pharmacology; D009838:Oligodeoxyribonucleotides / Q000494:pharmacology; D011994:Recombinant Proteins / Q000494:pharmacology; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013926:Thrombopoietin / Q000494:pharmacology
chemical_list: D003470:Culture Media; C508596:IL3 protein, human; D007377:Interleukin-3; D008565:Membrane Proteins; D009838:Oligodeoxyribonucleotides; D011994:Recombinant Proteins; C084881:flt3 ligand protein; D016899:Interferon-beta; D013926:Thrombopoietin; D015232:Dinoprostone