Abstract
We demonstrate that functional and phenotypic equivalents of mouse splenic CD8(+) and CD8(-) conventional dendritic cell (cDC) subsets can be generated in vitro when bone marrow is cultured with fms-like tyrosine kinase 3 (flt3) ligand. In addition to CD45RA(high) plasmacytoid DC, two distinct CD24(high) and CD11b(high) cDC subsets were present, and these subsets showed equivalent properties to splenic CD8(+) and CD8(-) cDC, respectively, in the following: 1) surface expression of CD11b, CD24, and signal regulatory protein-alpha; 2) developmental dependence on, and mRNA expression of, IFN regulatory factor-8; 3) mRNA expression of TLRs and chemokine receptors; 4) production of IL-12 p40/70, IFN-alpha, MIP-1alpha, and RANTES in response to TLR ligands; 5) expression of cystatin C; and 6) cross-presentation of exogenous Ag to CD8 T cells. Furthermore, despite lacking surface CD8 expression, the CD24(high) subset contained CD8 mRNA and up-regulated surface expression when transferred into mice. This culture system allows access to bona fide counterparts of the splenic DC subsets.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Animals
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Antigen Presentation / genetics
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Antigen Presentation / immunology
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Bone Marrow Cells / cytology
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Bone Marrow Cells / immunology*
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Bone Marrow Cells / metabolism
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CD4 Antigens / biosynthesis
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CD4 Antigens / genetics
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CD8 Antigens / biosynthesis*
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CD8 Antigens / genetics
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Cell Differentiation / genetics
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Cell Differentiation / immunology*
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Cells, Cultured
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Chemokines / biosynthesis
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Cross-Priming / genetics
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Cross-Priming / immunology
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Cystatin C
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Cystatins / biosynthesis
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Cytokines / biosynthesis
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Dendritic Cells / cytology*
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Dendritic Cells / immunology*
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Dendritic Cells / metabolism
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Immunophenotyping
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Interferon Regulatory Factors
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Ligands
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Membrane Glycoproteins / biosynthesis
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Membrane Proteins / metabolism*
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Mice
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Mice, Inbred C57BL
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Mice, Knockout
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Mice, Transgenic
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Proto-Oncogene Proteins / metabolism*
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Receptor Protein-Tyrosine Kinases / metabolism*
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Receptors, Cell Surface / biosynthesis
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Receptors, Chemokine / biosynthesis
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Repressor Proteins / biosynthesis
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Repressor Proteins / genetics
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Repressor Proteins / physiology
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Spleen / cytology
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Spleen / immunology*
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Spleen / metabolism
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Toll-Like Receptors
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fms-Like Tyrosine Kinase 3
Substances
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CD4 Antigens
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CD8 Antigens
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Chemokines
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Cst3 protein, mouse
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Cystatin C
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Cystatins
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Cytokines
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Interferon Regulatory Factors
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Ligands
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Membrane Glycoproteins
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Membrane Proteins
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Proto-Oncogene Proteins
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Receptors, Cell Surface
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Receptors, Chemokine
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Repressor Proteins
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Toll-Like Receptors
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flt3 ligand protein
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interferon regulatory factor-8
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Flt3 protein, mouse
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Receptor Protein-Tyrosine Kinases
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fms-Like Tyrosine Kinase 3