Skin Tregs generated early in life are crucial for adult ...
1
Results Skin Tregs generated early in life are crucial for adult skin homeostasis Inchul Cho 1,2 , Jessie Xu 1,2 , Niwa Ali 1,2 1. Centre for Stem Cells and Regenerative Medicine, UK; 2. The Francis Crick Institute, UK References 1. Ali, N., Zirak, B., Rodriguez, R., Pauli, M., Truong, H., Lai, K., Ahn, R., Corbin, K., Lowe, M., Scharschmidt, T., Taravati, K., Tan, M., Ricardo-Gonzalez, R., Nosbaum, A., Bertolini, M., Liao, W., Nestle, F., Paus, R., Cotsarelis, G., Abbas, A. and Rosenblum, M., 2017. Regulatory T Cells in Skin Facilitate Epithelial Stem Cell Differentiation. Cell, 169(6), pp.1119-1129.e11. 2. Mathur, A., Zirak, B., Boothby, I., Tan, M., Cohen, J., Mauro, T., Mehta, P., Lowe, M., Abbas, A., Ali, N. and Rosenblum, M., 2019. Treg-Cell Control of a CXCL5-IL-17 Inflammatory Axis Promotes Hair-Follicle-Stem-Cell Differentiation During Skin-Barrier Repair. Immunity, 50(3), pp.655-667.e4. Introduction Regulatory T cells (Tregs) govern hair growth and wound healing by controlling the activity of stem cells in the mouse skin 1,2 . However, their functions in the neonatal skin during postnatal development is unknown. Methods We use Foxp3-EGFP-DTR transgenic mice to deplete Tregs during early stages of postnatal skin development. The skin from Treg-depleted mice are profiled by flow cytometric profiling, histology and whole skin RNA-sequencing 1. Neonatal Tregs are required for adult skin homeostasis Conclusion 2. RNA-seq analysis suggests neuronal defect in the skin immediately after Treg depletion 3. Tregs are required during early phase of postnatal skin development DT PBS Ventral DT PBS PBS DT 0 10 20 30 40 Follicular density ✱ PBS DT 0 2000 4000 6000 8000 10000 PIgment index ✱ Treg-depleted Treg-sufficient DGE Analysis GO Analysis Late DT DT PBS Harvest P6 P8 P10 P12 P28 PBS PBS Harvest Early DT DT PBS Harvest Full DT Harvest DT Phenotype PBS Late DT Early DT Full DT Lack of pigmentation No Not much Yes Yes Less hair No Not much Yes Yes Scaling skin No No Not much Yes PBS Late DT Early DT Full DT 4. Early DT and Full DT groups have high effector T cell (Teff), Langerin- myeloid numbers and LC activation on postnatal day 28. PBS Early DT Late DT Full DT 0 2000 4000 6000 Teffs (Abs No) Absolute number (per 6 SQCM) ✱✱ ns ✱ PBS Early DT Late DT Full DT 0 100 200 300 400 Langerin- (Abs No) Absolute number (per 6 SQCM) ✱✱✱ ✱✱ ns ✱✱ PBS Early DT Late DT Full DT 4000 5000 6000 7000 8000 9000 MHC2 (MFI) ✱✱ ✱ ns PBS Early DT Late DT Full DT 1. Neonatal Tregs are required for pigmentation and maintenance of hair follicles 2. Depletion of Tregs causes immediate transcriptomic changes associated with neurobiology. 3. Absence of Tregs during P6-P8, but not P10-P12, causes skin defects. 4. Elevated Teff and Langerin- myeloid number, as well was activated LCs, may contribute to skin defects. 5. We will assess how cytokines, secreted by Teffs, affect neuronal innervation of hair follicles. Dorsal P6 P8 P10 P12 P28 Harvest LCs
Transcript of Skin Tregs generated early in life are crucial for adult ...
Results
Skin Tregs generated early in life arecrucial for adult skin homeostasis
Inchul Cho1,2, Jessie Xu1,2, Niwa Ali1,21. Centre for Stem Cells and Regenerative Medicine, UK; 2. The Francis Crick Institute, UK
References1. Ali, N., Zirak, B., Rodriguez, R., Pauli, M., Truong, H., Lai, K., Ahn, R., Corbin, K., Lowe, M., Scharschmidt, T., Taravati, K., Tan, M., Ricardo-Gonzalez, R., Nosbaum, A., Bertolini, M., Liao,
W., Nestle, F., Paus, R., Cotsarelis, G., Abbas, A. and Rosenblum, M., 2017. Regulatory T Cells in Skin Facilitate Epithelial Stem Cell Differentiation. Cell, 169(6), pp.1119-1129.e11.2. Mathur, A., Zirak, B., Boothby, I., Tan, M., Cohen, J., Mauro, T., Mehta, P., Lowe, M., Abbas, A., Ali, N. and Rosenblum, M., 2019. Treg-Cell Control of a CXCL5-IL-17 Inflammatory Axis
Promotes Hair-Follicle-Stem-Cell Differentiation During Skin-Barrier Repair. Immunity, 50(3), pp.655-667.e4.
IntroductionRegulatory T cells (Tregs) govern hair growth and woundhealing by controlling the activity of stem cells in the mouseskin1,2. However, their functions in the neonatal skin duringpostnatal development is unknown.
MethodsWe use Foxp3-EGFP-DTR transgenic mice to deplete Tregsduring early stages of postnatal skin development. The skinfrom Treg-depleted mice are profiled by flow cytometricprofiling, histology and whole skin RNA-sequencing
1. Neonatal Tregs are required for adult skinhomeostasis
Conclusion
2. RNA-seq analysis suggests neuronal defect inthe skin immediately after Treg depletion
3. Tregs are required during early phase ofpostnatal skin development
DT
PBS
Ventral
DT
PBS
PBS DT0
10
20
30
40
Folli
cula
r de
nsity
✱
PBS DT0
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10000
PIg
men
t ind
ex
✱
Treg-depleted Treg-sufficient
DGE Analysis GO Analysis
Late DT DTPBS
Harvest
P6 P8 P10 P12 P28
PBSPBS
Harvest
Early DT DT PBS
Harvest
Full DT
HarvestDT
Phenotype PBS Late DT Early DT Full DTLack of pigmentation No Not much Yes YesLess hair No Not much Yes YesScaling skin No No Not much Yes
PBS Late DT
Early DT Full DT
4. Early DT and Full DT groups have high effector Tcell (Teff), Langerin- myeloid numbers and LCactivation on postnatal day 28.
PBS
Early
DT
Late D
T
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CD3- (Abs No)
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olut
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mbe
r (p
er 6
SQ
CM
) ns
PBS
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DT
Late D
T
Full DT
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Teffs (Abs No)
Abs
olut
e nu
mbe
r (p
er 6
SQ
CM
)
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ns
✱
PBS
Early
DT
Late D
T
Full DT
0
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DETCs (Abs No)
Abs
olut
e nu
mbe
r (p
er 6
SQ
CM
) ✱
✱
PBS
Early
DT
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T
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Tregs (Abs No)
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olut
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mbe
r (p
er 6
SQ
CM
)
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PBS
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Late D
T
Full DT
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dGDTCs (Abs No)
Abs
olut
e nu
mbe
r (p
er 6
SQ
CM
)
ns
PBS
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DT
Late D
T
Full DT
0
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CD8+ (Abs No)
Abs
olut
e nu
mbe
r (p
er 6
SQ
CM
)
✱
✱
✱✱✱✱
PBS
Early
DT
Late D
T
Full DT
0
100
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Langerin- (Abs No)
Abs
olut
e nu
mbe
r (p
er 6
SQ
CM
)
✱✱✱
✱✱
ns
✱✱
PBS
Early
DT
Late D
T
Full DT
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10000
20000
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DETCs (Abs No)
Abs
olut
e nu
mbe
r (p
er 6
SQ
CM
)
ns
PBS
Early
DT
Late D
T
Full DT
0
5000
10000
15000
20000
LCs (Abs No)
Abs
olut
e nu
mbe
r (p
er 6
SQ
CM
)
ns
PBS
Early
DT
Late D
T
Full DT
0
200
400
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CD4+ (Abs No)
Abs
olut
e nu
mbe
r (p
er 6
SQ
CM
)
✱✱✱
✱✱
✱✱✱
PBS
Early
DT
Late D
T
Full DT
0
1000
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ILCs (Abs No)
Abs
olut
e nu
mbe
r (p
er 6
SQ
CM
)
✱✱ ✱✱
✱
PBS
Early
DT
Late D
T
Full DT
0
1000
2000
3000
CD8+ (Abs No)
Abs
olut
e nu
mbe
r (p
er 6
SQ
CM
)
✱✱✱✱
✱✱✱
✱✱✱✱
PBS
Early
DT
Late D
T
Full DT
4000
5000
6000
7000
8000
9000
MH
C2
(MFI
)
✱✱ ✱
ns
PBS
Early DT
Late DT
Full DT
1. Neonatal Tregs are required for pigmentation and maintenance of hair follicles2. Depletion of Tregs causes immediate transcriptomic changes associated with neurobiology.3. Absence of Tregs during P6-P8, but not P10-P12, causes skin defects.4. Elevated Teff and Langerin- myeloid number, as well was activated LCs, may contribute to skin defects.5. We will assess how cytokines, secreted by Teffs, affect neuronal innervation of hair follicles.
DorsalP6 P8 P10 P12 P28
Harvest
LCs
