Harrisonet al.,Science 363 , eaat6280 (2019) 4 January 2019 3of11
0.7±0.30.8±0.351±5RORγtIsotype48±59±25±2RORγtGATA-3A% maxG ATA - 3S. epidermidis C. albicans
Iso
Th17Iso
Th17S. epidermidis - CD8+ T cellsBFoxp3 DAPI CD8αCSki
n
VA
T
LungcLPMusclesiLP
ThymusMATSLN
SpleenPPPLNLiverBloodMLNBM020406080100
GATA-3RORγt
T-bet***Foxp3+ LDTF+ (%)D21±3 13±58±427±4 36±53±2Foxp3YFP-CreGata3fl/wtFoxp3YFP-CreGata3fl/flFoxp3CD25EMLNSLNPLN
SpleenFoxp3YFP-Cre
Gata3fl/wtFoxp3YFP-Cre
Gata3fl/flF G051015
**CD4+ IL-5+ (%)Skin051015
**CD4+ IL-13+ (%)Skin
Foxp3YFP-CreGata3fl/wt Foxp3YFP-CreGata3fl/flHEosinophils (%)^10
020304050
**Skin051015Basophils (%)**Skin^0100200300050100Age (days)Skin inflammationpenetrance (%)Foxp3YFP-CreGata3fl/flFoxp3YFP-CreGata3fl/wtIJ
Foxp3YFP-CreGata3fl/wtFoxp3YFP-CreGata3fl/fl20 ±3 0.4±0.33 ±126 ±5 7 ±23 ±2Foxp3YFP-CreGata3fl/wtFoxp3YFP-CreGata3fl/flCD8+ IL-5+ (x103 )Naïve S. epi****00.51.01.52.02.5CD8+ IL13+ (x103 )Naïve S. epi****00.51.01.52.02.5KCD8βIL-5L
Foxp3YFP-CreGata3fl/wt Foxp3YFP-CreGata3fl/fl95±44±2HostBowieTgFoxp3YFP-CreGata3fl/wtFoxp3YFP-CreGata3fl/flCCR6IL-5RecipientBowieDonorTg51±5 0.6±0.30.3±0.154±4 0.5±0.20.3±0.152±6 5±20.5±0.350±7 4±30.2±0.1CD8+T cellsIL-1344±50.5±0.40.7±0.5 46±60.7±0.54±349±70.7±0.40.5±0.4 48±90.3±0.14±2CCR6Foxp3YFP-CreGata3fl/wtFoxp3YFP-CreGata3fl/flRecipientBowieDonorTgMFig. 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)].RESEARCH | RESEARCH ARTICLE
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