Harrisonet al.,Science 363 , eaat6280 (2019) 4 January 2019 3of11
0.7±0.3
0.8±0.3
51±5
ROR
γt
Isotype
48±5
9±2
5±2
ROR
γt
GATA-3
A
% max
G ATA - 3
S. epidermidis C. albicans
Iso
Th17
Iso
Th17
S. epidermidis - CD8+ T cells
B
Foxp3 DAPI CD8α
C
Ski
n
VA
T
LungcLPMusclesiLP
Thymus
MATSL
N
Spleen
PPPLNLiverBloodMLNBM
0
20
40
60
80
100
GATA-3
RORγt
T-bet
***
Foxp3
+ LDTF
+ (%)
D
21±3 13±5
8±4
27±4 36±5
3±2
Foxp3YFP-CreGata3fl/wtFoxp3YFP-CreGata3fl/fl
Foxp3
CD25
E
MLN
SLN
PLN
Spleen
Foxp3YFP-Cre
Gata3fl/wt
Foxp3YFP-Cre
Gata3fl/fl
F G
0
5
10
15
**
CD4
+ IL-5
+ (%)
Skin
0
5
10
15
**
CD4
+ IL-13
+ (%)
Skin
Foxp3YFP-CreGata3fl/wt Foxp3YFP-CreGata3fl/fl
H
Eosinophils (%)^10
0
20
30
40
50
**
Skin
0
5
10
15
Basophils (%)
**
Skin^0100200300
0
50
100
Age (days)
Skin inflammationpenetrance (%)
Foxp3YFP-CreGata3fl/fl
Foxp3YFP-CreGata3fl/wt
I
J
Foxp3YFP-CreGata3fl/wt
Foxp3YFP-CreGata3fl/fl
20 ±3 0.4±0.3
3 ±1
26 ±5 7 ±2
3 ±2
Foxp3YFP-CreGata3fl/wtFoxp3YFP-CreGata3fl/fl
CD8
+ IL-5
+ (x10
3 )
Naïve S. epi
*
***
0
0.5
1.0
1.5
2.0
2.5
CD8
+ IL13
+ (x10
3 )
Naïve S. epi
*
***
0
0.5
1.0
1.5
2.0
2.5
K
CD8
β
IL-5
L
Foxp3YFP-CreGata3fl/wt Foxp3YFP-CreGata3fl/fl
95±4
4±2
Host
BowieTg
Foxp3YFP-CreGata3fl/wtFoxp3YFP-CreGata3fl/fl
CCR6
IL-5
Recipient
Bowie
DonorTg
51±5 0.6±0.3
0.3±0.1
54±4 0.5±0.2
0.3±0.1
52±6 5±2
0.5±0.3
50±7 4±3
0.2±0.1
CD8+T cells
IL-13
44±5
0.5±0.4
0.7±0.5 46±6
0.7±0.5
4±3
49±7
0.7±0.4
0.5±0.4 48±9
0.3±0.1
4±2
CCR6
Foxp3YFP-CreGata3fl/wtFoxp3YFP-CreGata3fl/fl
Recipient
Bowie
DonorTg
M
Fig. 2. Local defects
in immunoregulation
unleash type 2
immunity from
commensal-specific
T cells.(A)Represent-
ative contour plots of
RORgtandGATA-3
expression by skin
CD8+T cells from
S. epidermidis–colonized
wild-type mice. (B)Rep-
resentative histogram
plots of GATA-3
expression by RORgt+
CD4+TH17 cells from
the skin of commensal-
colonized wild-type
mice. (C) Representa-
tive confocal imaging
volume-projected
along thezaxis of
epidermal skin from
S. epidermidis–colonized
Foxp3gfpmice. (D)Fre-
quencies of Foxp3+Treg
cells coexpressing
lineage-defining
transcription factors
(LDTFs) within indicated
tissues of naïve wild-
type mice. VAT, visceral
adipose tissue; cLP,
colonic lamina propria;
siLP, small intestinal
lamina propria; MAT,
mesenteric adipose
tissue; SLN, skin-
draining lymph node;
PP, Peyer’spatch;PLN,
para-aortic lymph
node; MLN, mesenteric
lymph node; BM, bone
marrow. (E)Represent-
ative contour plots of
Foxp3 and CD25
expression by skin
CD4+T cells from naïve
Foxp3YFP-CreGata3fl/wt
andFoxp3YFP-CreGata3fl/fl
mice. (F)Representative
cutaneous lymphadeno-
pathy inFoxp3YFP-Cre-
Gata3fl/flcompared to
Foxp3YFP-CreGata3fl/wt
control mice. (G)Fre-
quencies of IL-5–and IL-13–producing skin CD4+T cells from naïve
Foxp3YFP-CreGata3fl/wtandFoxp3YFP-CreGata3fl/flmice. (H) Frequencies
of skin eosinophils and basophils from naïveFoxp3YFP-CreGata3fl/wtand
Foxp3YFP-CreGata3fl/flmice. (I) Cumulative incidence of skin inflammation
among naïveFoxp3YFP-CreGata3fl/wtandFoxp3YFP-CreGata3fl/flmice.
(J) Representative histological micrograph of skin tissue from naïve
Foxp3YFP-CreGata3fl/wtandFoxp3YFP-CreGata3fl/flmice. Scale bars, 250mm.
(K) Representative contour plots of CD8bexpression and IL-5 production
potential by TCRb+T cells from the skin ofS. epidermidis–colonized
Foxp3YFP-CreGata3fl/wtandFoxp3YFP-CreGata3fl/flmice. (L) Total numbers
of IL-5–and IL-13–producing CD8+Tcells from the skin ofS. epidermidis–
colonizedFoxp3YFP-CreGata3fl/wtandFoxp3YFP-CreGata3fl/flmice.
(M) Representative contour plots of IL-5 and IL-13 production by CD8+Tcells in
Foxp3YFP-CreGata3fl/wtandFoxp3YFP-CreGata3fl/flmice adoptively transferred
with BowieTgT cells before colonization withS. epidermidis.Numbersin
representative plots indicate means ± SD. Each dot represents an individual
mouse. Data represent at least two experiments with three to seven mice per
group. Cumulative skin inflammation data (I) represent 25 mice per genotype.
*P<0.05,**P< 0.01, ***P< 0.001 as calculated using Studentttest [(G), (H)]
or one-way ANOVA with Holm-Šidák multiple comparison test [(D), (L)].
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