title: Murine plasmacytoid pre-dendritic cells generated from Flt3 ligand-supplemented bone marrow cultures are immature APCs.
abstract: The putative counterparts of human plasmacytoid pre-dendritic cells (pDCs) have been described in vivo in mouse models and very recently in an in vitro culture system. In this study, we report that large numbers of bone marrow-derived murine CD11c(+)B220(+) pDCs can be generated with Flt3 ligand (FL) as the sole exogenous differentiation/growth factor and that pDC generation is regulated in vivo by FL because FL-deficient mice showed a major reduction in splenic pDC numbers. We extensively analyzed bone marrow-derived CD11c(+)B220(+) pDCs and described their immature APC phenotype based on MHC class II, activation markers, and chemokine receptor level of expression. CD11c(+)B220(+) pDCs showed a nonoverlapping Toll-like receptor pattern of expression distinct from that of classical CD11c(+)B220(-) dendritic cells and were poor T cell stimulators. Stimulation of CD11c(+)B220(+) pDCs with oligodeoxynucleotides containing certain CpG motifs plus CD40 ligand plus GM-CSF led to increased MHC class II, CD80, CD86, and CD8alpha expression levels, to a switch in chemokine receptor expression that affected their migration, to IFN-alpha and IL-12 secretion, and to the acquisition of priming capacities for both CD4(+) and CD8(+) OVA-specific TCR-transgenic naive T cells. Thus, the in vitro generation of murine pDCs may serve as a useful tool to further investigate pDC biology as well as the potential role of these cells in viral immunity and other settings.
authors.affiliation: Amgen Inc., Seattle, WA 98101, USA. brawandp@amgen.com
authors.forename: Pierre
authors.initials: P
authors.lastname: Brawand
authors.forename: David R
authors.initials: DR
authors.lastname: Fitzpatrick
authors.forename: Brad W
authors.initials: BW
authors.lastname: Greenfield
authors.forename: Kenneth
authors.initials: K
authors.lastname: Brasel
authors.forename: Charles R
authors.initials: CR
authors.lastname: Maliszewski
authors.forename: Thibaut
authors.initials: T
authors.lastname: De Smedt
mesh_terms: D000818:Animals; D000938:Antigen-Presenting Cells / Q000166:cytology* / Q000276:immunology / Q000378:metabolism; D001854:Bone Marrow Cells / Q000166:cytology* / Q000276:immunology / Q000378:metabolism; D039521:CD11c Antigen / Q000096:biosynthesis; D016827:CD8 Antigens / Q000096:biosynthesis; D002454:Cell Differentiation / Q000235:genetics / Q000276:immunology; D002455:Cell Division / Q000235:genetics / Q000276:immunology; D002465:Cell Movement / Q000276:immunology; D002478:Cells, Cultured; D017077:Culture Media, Conditioned / Q000494:pharmacology; D003713:Dendritic Cells / Q000166:cytology* / Q000276:immunology / Q000378:metabolism; D029721:Drosophila Proteins; D005260:Female; D006133:Growth Substances / Q000172:deficiency / Q000235:genetics / Q000502:physiology*; D016130:Immunophenotyping; D016898:Interferon-alpha / Q000378:metabolism; D018664:Interleukin-12 / Q000378:metabolism; D017493:Leukocyte Common Antigens / Q000096:biosynthesis; D008562:Membrane Glycoproteins / Q000096:biosynthesis; D008565:Membrane Proteins / Q000172:deficiency / Q000235:genetics / Q000502:physiology*; D051379:Mice; D008807:Mice, Inbred BALB C; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D008822:Mice, Transgenic; D011956:Receptors, Cell Surface / Q000096:biosynthesis; D013234:Stem Cells / Q000166:cytology* / Q000276:immunology* / Q000378:metabolism; D051193:Toll-Like Receptors
chemical_list: D039521:CD11c Antigen; D016827:CD8 Antigens; C431698:CD8alpha antigen; D017077:Culture Media, Conditioned; D029721:Drosophila Proteins; D006133:Growth Substances; D016898:Interferon-alpha; D008562:Membrane Glycoproteins; D008565:Membrane Proteins; D011956:Receptors, Cell Surface; D051193:Toll-Like Receptors; C084881:flt3 ligand protein; D018664:Interleukin-12; D017493:Leukocyte Common Antigens