Cutting edge: generation of splenic CD8+ and CD8- dendritic cell equivalents in Fms-like tyrosine kinase 3 ligand bone marrow cultures.
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.
Naik
Shalin H
SH
Immunology Division and the Cooperative Research Centre for Vaccine Technology, Walter and Eliza Hall Institute of Medical Research, Melbourne, Australia. naik@wehi.edu.au
Proietto
Anna I
AI
Wilson
Nicholas S
NS
Dakic
Aleksandar
A
Schnorrer
Petra
P
Fuchsberger
Martina
M
Lahoud
Mireille H
MH
O'Keeffe
Meredith
M
Shao
Qi-xiang
QX
Chen
Wei-feng
WF
Villadangos
José A
JA
Shortman
Ken
K
Wu
Li
L
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
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