title: Cutting edge: generation of splenic CD8+ and CD8- dendritic cell equivalents in Fms-like tyrosine kinase 3 ligand bone marrow cultures.
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.
authors.affiliation: Immunology Division and the Cooperative Research Centre for Vaccine Technology, Walter and Eliza Hall Institute of Medical Research, Melbourne, Australia. naik@wehi.edu.au
authors.forename: Shalin H
authors.initials: SH
authors.lastname: Naik
authors.forename: Anna I
authors.initials: AI
authors.lastname: Proietto
authors.forename: Nicholas S
authors.initials: NS
authors.lastname: Wilson
authors.forename: Aleksandar
authors.initials: A
authors.lastname: Dakic
authors.forename: Petra
authors.initials: P
authors.lastname: Schnorrer
authors.forename: Martina
authors.initials: M
authors.lastname: Fuchsberger
authors.forename: Mireille H
authors.initials: MH
authors.lastname: Lahoud
authors.forename: Meredith
authors.initials: M
authors.lastname: O'Keeffe
authors.forename: Qi-xiang
authors.initials: QX
authors.lastname: Shao
authors.forename: Wei-feng
authors.initials: WF
authors.lastname: Chen
authors.forename: José A
authors.initials: JA
authors.lastname: Villadangos
authors.forename: Ken
authors.initials: K
authors.lastname: Shortman
authors.forename: Li
authors.initials: L
authors.lastname: Wu
mesh_terms: D000818:Animals; D017951:Antigen Presentation / Q000235:genetics / Q000276:immunology; D001854:Bone Marrow Cells / Q000166:cytology / Q000276:immunology* / Q000378:metabolism; D015704:CD4 Antigens / Q000096:biosynthesis / Q000235:genetics; D016827:CD8 Antigens / Q000096:biosynthesis* / Q000235:genetics; D002454:Cell Differentiation / Q000235:genetics / Q000276:immunology*; D002478:Cells, Cultured; D018925:Chemokines / Q000096:biosynthesis; D045142:Cross-Priming / Q000235:genetics / Q000276:immunology; D055316:Cystatin C; D015891:Cystatins / Q000096:biosynthesis; D016207:Cytokines / Q000096:biosynthesis; D003713:Dendritic Cells / Q000166:cytology* / Q000276:immunology* / Q000378:metabolism; D016130:Immunophenotyping; D050835:Interferon Regulatory Factors; D008024:Ligands; D008562:Membrane Glycoproteins / Q000096:biosynthesis; D008565:Membrane Proteins / Q000378:metabolism*; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D008822:Mice, Transgenic; D011518:Proto-Oncogene Proteins / Q000378:metabolism*; D020794:Receptor Protein-Tyrosine Kinases / Q000378:metabolism*; D011956:Receptors, Cell Surface / Q000096:biosynthesis; D019707:Receptors, Chemokine / Q000096:biosynthesis; D012097:Repressor Proteins / Q000096:biosynthesis / Q000235:genetics / Q000502:physiology; D013154:Spleen / Q000166:cytology / Q000276:immunology* / Q000378:metabolism; D051193:Toll-Like Receptors; D051941:fms-Like Tyrosine Kinase 3; D000099185:Interferon Regulatory Factor-8
chemical_list: D015704:CD4 Antigens; D016827:CD8 Antigens; D018925:Chemokines; D055316:Cystatin C; D015891:Cystatins; D016207:Cytokines; D050835:Interferon Regulatory Factors; D008024:Ligands; D008562:Membrane Glycoproteins; D008565:Membrane Proteins; D011518:Proto-Oncogene Proteins; D020794:Receptor Protein-Tyrosine Kinases; D011956:Receptors, Cell Surface; D019707:Receptors, Chemokine; D012097:Repressor Proteins; D051193:Toll-Like Receptors; D051941:fms-Like Tyrosine Kinase 3; D000099185:Interferon Regulatory Factor-8; C525031:Cst3 protein, mouse; C084881:flt3 ligand protein; C497971:Flt3 protein, mouse