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CUTTING EDGE |
* Immunology Division and the Cooperative Research Centre for Vaccine Technology, Walter and Eliza Hall Institute of Medical Research, Melbourne, Australia; Now at Bavarian Nordic, Munich, Germany; and Department of Immunology, Peking University Health Science Center, Beijing, China
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 CD45RAhigh plasmacytoid DC, two distinct CD24high and CD11bhigh 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-
; 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-, MIP-1, 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 CD24high 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.
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