1. +/- antigen specific T cells
A) Antigen targeting to murine DC subpopulations induces different T cell responses in vivo
CD8+
T cells
CD11c+CD8+
Lehmann et al., Vaccines, 2016
Heidkamp et al., Encycl. of Cell Biology, 2016
Neubert et al., JI, 2014
Eissing et al., Immunoly Letters, 2014
Milo et al., Blood, 2013
Lindmark et al., JAutoimmun, 2012
Loschko et al., JI,
Loschko et al., JI,
O’Kamphorst et al., JI, 2010
Do et al., EJI, 2010
Yamazaki and Dudziak, JI, 2008
Dudziak et al., Science, 2007
Soares et al., JEM, 2007
Shakhar et al., Nat. Immunol. 2005
Lindquist et al., Nat. Immunol., 2004
DEC205-Ova
B220 DEC205 DCIR2
CD4+
T cells
CD11c+CD8-
Spleen
DCIR2-Ova (33D1)
1. Tumor responses?
2. APC presentation?
3. Stimulations differences?
4. Human?
B) Antigen presented by DC subpopulations leads to differentiation and expansion of antigen specific regulatory T cells in the steady state in vivo
Overview adapted from Gorelik and Flavell,
Nat. Rev. Immunol, 2002
1. Direct effect of TGF-β on self-reactive T cell
2. Indirect effect of TGF-β via Treg cells
3. Generation of regulatory DC via Treg cells
TGF-b
TGF-β -BP2
TGF-β Receptor 1
SMAD1
SMAD4
CD8–
CD8+ 2
SpDC
PBS
33D1-OVA
DEC-OVA
CFSE
FoxP3
0.25
2.0 95
3.0
2.5
0.26 97
4.5 16 22 58
CD4+CD25- T cells → CD4+CD25+FoxP3+ Treg
Dependend on CD8+ DCs, TGF-β
Expansion of FoxP3+ T cells by CD8- DCs
FoxP3+
expansion
FoxP3-
FoxP3+
TGF-β
CD8+ DCs
CD8- DCs
5. Differentiation of CD4+ naïve T cells into Treg cells
by simultaneous antigen presentation and secretion
of TGF-β by CD8+DCs
6. Expansion of Treg cells by antigen presenting CD8- DCs
(maybe also induction of regulatory DC)
Yamazaki, Dudziak et al., et al., JI, 2008
Experimental Design
TLR-Ligand
C57BL/6 WT
CD11c-YFP
TLR-/-, MyD88-/-
CD11c+CD8+
CD11c+CD8-
pDC
OT-I, OT-II
In vitro
In vivo
microarray and miRNA
protein changes (WB)
DC migration (Immunfluorescence, 2- Photon)
T cell proliferation (in vivo und in vitro)
memory (in vivo und in vitro)
antigen processing machinery
Injection of
TLR-ligands
+/- antigen specific T cells
Immature DC
Mature DC
C) Molecular analysis of DC subpopulations and their interacting T cells
B220/DCIR2/DEC205 B T
Baranska et al., in preparation