title: Selective and efficient generation of functional Batf3-dependent CD103+ dendritic cells from mouse bone marrow.
abstract: Multiple subsets of FMS-like tyrosine kinase 3 ligand (FLT3L)-dependent dendritic cells (DCs) control T-cell tolerance and immunity. In mice, Batf3-dependent CD103(+) DCs efficiently enter lymph nodes and cross-present antigens, rendering this conserved DC subset a promising target for tolerance induction or vaccination. However, only limited numbers of CD103(+) DCs can be isolated with current methods. Established bone marrow culture protocols efficiently generate monocyte-derived DCs or produce a mixture of FLT3L-dependent DC subsets. We show that CD103(+) DC development requires prolonged culture time and continuous action of both FLT3L and granulocyte macrophage colony-stimulating factor (GM-CSF), explained by a dual effect of GM-CSF on DC precursors and differentiating CD103(+) DCs. Accordingly, we established a novel method to generate large numbers of CD103(+) DCs (iCD103-DCs) with limited presence of other DC subsets. iCD103-DCs develop in a Batf3- and Irf8-dependent fashion, express a CD8α/CD103 DC gene signature, cross-present cell-associated antigens, and respond to TLR3 stimulation. Thus, iCD103-DCs reflect key features of tissue CD103(+) DCs. Importantly, iCD103-DCs express high levels of CCR7 upon maturation and migrate to lymph nodes more efficiently than classical monocyte-derived DCs. Finally, iCD103-DCs induce T cell-mediated protective immunity in vivo. Our study provides insights into CD103(+) DC development and function.
authors.affiliation: Institute of Infection Immunology, TWINCORE, Centre for Experimental and Clinical Infection Research, Medical School Hannover and the Helmholtz Centre for Infection Research, Hannover, Germany; and.
authors.forename: Christian Thomas
authors.initials: CT
authors.lastname: Mayer
authors.affiliation: Institute of Infection Immunology, TWINCORE, Centre for Experimental and Clinical Infection Research, Medical School Hannover and the Helmholtz Centre for Infection Research, Hannover, Germany; and.
authors.forename: Peyman
authors.initials: P
authors.lastname: Ghorbani
authors.affiliation: Institute of Infection Immunology, TWINCORE, Centre for Experimental and Clinical Infection Research, Medical School Hannover and the Helmholtz Centre for Infection Research, Hannover, Germany; and.
authors.forename: Amrita
authors.initials: A
authors.lastname: Nandan
authors.affiliation: Institute of Infection Immunology, TWINCORE, Centre for Experimental and Clinical Infection Research, Medical School Hannover and the Helmholtz Centre for Infection Research, Hannover, Germany; and.
authors.forename: Markus
authors.initials: M
authors.lastname: Dudek
authors.affiliation: Institute of Infection Immunology, TWINCORE, Centre for Experimental and Clinical Infection Research, Medical School Hannover and the Helmholtz Centre for Infection Research, Hannover, Germany; and.
authors.forename: Catharina
authors.initials: C
authors.lastname: Arnold-Schrauf
authors.affiliation: Institute of Infection Immunology, TWINCORE, Centre for Experimental and Clinical Infection Research, Medical School Hannover and the Helmholtz Centre for Infection Research, Hannover, Germany; and.
authors.forename: Christina
authors.initials: C
authors.lastname: Hesse
authors.affiliation: Institute of Infection Immunology, TWINCORE, Centre for Experimental and Clinical Infection Research, Medical School Hannover and the Helmholtz Centre for Infection Research, Hannover, Germany; and.
authors.forename: Luciana
authors.initials: L
authors.lastname: Berod
authors.affiliation: Institute of Infection Immunology, TWINCORE, Centre for Experimental and Clinical Infection Research, Medical School Hannover and the Helmholtz Centre for Infection Research, Hannover, Germany; and.
authors.forename: Philipp
authors.initials: P
authors.lastname: Stüve
authors.affiliation: Institute of Infection Immunology, TWINCORE, Centre for Experimental and Clinical Infection Research, Medical School Hannover and the Helmholtz Centre for Infection Research, Hannover, Germany; and.
authors.forename: Franz
authors.initials: F
authors.lastname: Puttur
authors.affiliation: Department of Oncological Sciences, Mount Sinai School of Medicine, New York, NY.
authors.forename: Miriam
authors.initials: M
authors.lastname: Merad
authors.affiliation: Institute of Infection Immunology, TWINCORE, Centre for Experimental and Clinical Infection Research, Medical School Hannover and the Helmholtz Centre for Infection Research, Hannover, Germany; and.
authors.forename: Tim
authors.initials: T
authors.lastname: Sparwasser
mesh_terms: D000818:Animals; D015703:Antigens, CD / Q000032:analysis / Q000276:immunology*; D050976:Basic-Leucine Zipper Transcription Factors / Q000032:analysis / Q000276:immunology*; D001854:Bone Marrow Cells / Q000166:cytology*; D018929:Cell Culture Techniques / Q000379:methods*; D002454:Cell Differentiation; D002478:Cells, Cultured; D003713:Dendritic Cells / Q000166:cytology* / Q000276:immunology*; D016178:Granulocyte-Macrophage Colony-Stimulating Factor / Q000276:immunology; D007111:Immunity, Cellular; D039001:Integrin alpha Chains / Q000032:analysis / Q000276:immunology*; D008565:Membrane Proteins / Q000276:immunology; D051379:Mice; D012097:Repressor Proteins / Q000032:analysis / Q000276:immunology*; D013601:T-Lymphocytes / Q000276:immunology; D051196:Toll-Like Receptor 3 / Q000276:immunology
chemical_list: D015703:Antigens, CD; D050976:Basic-Leucine Zipper Transcription Factors; D039001:Integrin alpha Chains; D008565:Membrane Proteins; D012097:Repressor Proteins; C533289:SNFT protein, mouse; D051196:Toll-Like Receptor 3; C091367:alpha E integrins; C084881:flt3 ligand protein; D016178:Granulocyte-Macrophage Colony-Stimulating Factor
references.citation: Nat Immunol. 2006 Jun;7(6):663-71
references.pmid: 16680143
references.citation: J Immunol. 2009 Aug 1;183(3):1514-7
references.pmid: 19570827
references.citation: J Immunol Methods. 1999 Feb 1;223(1):77-92
references.pmid: 10037236
references.citation: J Immunol. 2011 Feb 15;186(4):2422-9
references.pmid: 21242525
references.citation: J Exp Med. 2010 Jun 7;207(6):1261-71
references.pmid: 20479117
references.citation: J Exp Med. 2009 Dec 21;206(13):3115-30
references.pmid: 20008528
references.citation: Cell Mol Immunol. 2012 Nov;9(6):446-54
references.pmid: 23085949
references.citation: Science. 2008 Nov 14;322(5904):1097-100
references.pmid: 19008445
references.citation: EMBO J. 2011 May 17;30(13):2690-704
references.pmid: 21587207
references.citation: Nat Rev Immunol. 2006 Apr;6(4):295-307
references.pmid: 16557261
references.citation: Blood. 2012 Jun 21;119(25):6052-62
references.pmid: 22442345
references.citation: Blood. 2009 Apr 9;113(15):3418-27
references.pmid: 19176316
references.citation: Blood. 2009 Jul 23;114(4):835-43
references.pmid: 19465690
references.citation: J Immunol. 2010 Sep 15;185(6):3313-25
references.pmid: 20702727
references.citation: Eur J Immunol. 2011 Sep;41(9):2585-95
references.pmid: 21660938
references.citation: J Immunol. 2011 May 1;186(9):5184-92
references.pmid: 21422244
references.citation: J Clin Invest. 2012 Nov;122(11):4037-47
references.pmid: 23041628
references.citation: Nat Rev Cancer. 2012 Mar 22;12(4):265-77
references.pmid: 22437871
references.citation: Immunity. 2012 Jun 29;36(6):1031-46
references.pmid: 22749353
references.citation: Annu Rev Immunol. 2013;31:563-604
references.pmid: 23516985
references.citation: Immunity. 2004 Aug;21(2):279-88
references.pmid: 15308107
references.citation: J Immunol. 2008 Nov 15;181(10):6923-33
references.pmid: 18981112
references.citation: Methods Mol Biol. 2010;595:177-86
references.pmid: 19941112
references.citation: Immunity. 2009 Sep 18;31(3):513-25
references.pmid: 19733489
references.citation: J Exp Med. 2005 Oct 17;202(8):1051-61
references.pmid: 16216886
references.citation: Nat Immunol. 2012 Nov;13(11):1118-28
references.pmid: 23023392
references.citation: Proc Natl Acad Sci U S A. 2004 Jun 15;101(24):8981-6
references.pmid: 15184678
references.citation: J Exp Med. 2008 Nov 24;205(12):2839-50
references.pmid: 18981237
references.citation: J Exp Med. 2003 Aug 18;198(4):615-21
references.pmid: 12925677
references.citation: Immunol Rev. 2010 Mar;234(1):45-54
references.pmid: 20193011
references.citation: J Immunol. 2001 May 1;166(9):5448-55
references.pmid: 11313382
references.citation: J Immunol. 2010 Feb 15;184(4):2116-23
references.pmid: 20083654
references.citation: J Exp Med. 2010 Jun 7;207(6):1283-92
references.pmid: 20479118
references.citation: Nat Immunol. 2012 Dec;13(12):1145-54
references.pmid: 23160217
references.citation: Cell. 2013 Aug 15;154(4):843-58
references.pmid: 23953115
references.citation: J Immunol. 2005 Jun 1;174(11):6592-7
references.pmid: 15905497
references.citation: J Immunol. 2008 Mar 1;180(5):3019-27
references.pmid: 18292524
references.citation: PLoS One. 2011;6(10):e25660
references.pmid: 22065991
references.citation: J Immunol. 2005 May 1;174(9):5444-55
references.pmid: 15845457
references.citation: Eur J Immunol. 1998 Jun;28(6):2045-54
references.pmid: 9645386
references.citation: Nat Immunol. 2009 May;10(5):488-95
references.pmid: 19349986
references.citation: Nat Immunol. 2012 Sep;13(9):888-99
references.pmid: 22797772
references.citation: Front Immunol. 2012 Jul 03;3:183
references.pmid: 22783257
references.citation: Mucosal Immunol. 2012 Mar;5(2):150-60
references.pmid: 22166938
references.citation: J Immunol. 2004 Aug 1;173(3):1826-33
references.pmid: 15265914
references.citation: Nat Rev Immunol. 2007 Jan;7(1):19-30
references.pmid: 17170756
references.citation: J Exp Med. 2007 Aug 6;204(8):1757-64
references.pmid: 17620361
references.citation: J Immunol. 2007 Feb 15;178(4):2000-7
references.pmid: 17277103
references.citation: J Exp Med. 2010 Jan 18;207(1):189-206
references.pmid: 20038600
references.citation: J Immunol. 2008 Nov 1;181(9):6178-88
references.pmid: 18941208