Science - USA (2019-01-04)

Harrisonet al.,Science 363 , eaat6280 (2019) 4 January 2019 2of11

IL-17A

IFN-γ

A

C

E

Polyclonal

CD8+ T cells

23±3

22±5

BowieTg T cells

20±4

24±2

18±2

26±4

f-MIIINA:H2-M3+ T cells

CD103

CD69

f-MIIINA:H2-M3+ T cells

7±2 88±2

0 7±1

Spleen

f-MIIINA:H2-M3SLNSkin

+ (% Host)

0

25

50

75

100

D

CD103

CD69

Polyclonal

CD8+ T cells f-MIIINA:H2-M3+ T cells

18±2 66±6

10±4

BowieTg T cells

19±3 73±8

7±5

11±1 72±5

12±4

FG

ROR

γt

CCR6

47±5

Tc17

CD8+ T cells

16±5

34±8

IL-17A

CCR6

CD8+ T cells

I

CCR6

IL-13

43±6 0.2±0.1

0.1±0.1

41±4 2±1

0.4±0.2

CCR6

IL-5

i.d. PBS i.d. Chitin

41±7 0.4±0.3

0.2±0.1

43±5 4±2

0.4±0.3

H

0

10

20

30

Ctrl

i.d. PBSi.d. ChitinSand fly

Ctrl

i.d. PBSi.d. ChitinSand fly

IFN-

+γ

(%)

Tc1 Tc17

0

2

4

6

8

10

Ctrl

i.d. PBSi.d. ChitinSand fly

Ctrl

i.d. PBSi.d. ChitinSand fly

IL-5

+ (%)

**

**

Tc1 Tc17

0

2

4

6

IL-13

+ (%)

Ctrl

i.d. PBSi.d. ChitinSand fly

Ctrl

i.d. PBSi.d. ChitinSand fly

**

**

Tc1 Tc17

B

CD49f DAPI CD8a CD45.1 (BowieTg)

150μm

CD8α CD45.1 CD8α CD45.1

50μm

0

10

20

30

Ctrl

i.d. PBSi.d. ChitinSand fly

Ctrl

i.d. PBSi.d. ChitinSand fly

Tc1 Tc17

IL-17A

+ (%)

CD8+ T cells

i.d. PBS i.d. Chitin

CD8+ T cells

2±1 2±1 3±1

45±4

41±5

ROR

γt

T-bet

CD8+ T cells

Tc17

Tc1

Fig. 1. Acute injury licenses type 2
cytokine production from commensal-
specific type 17 cells.(AtoC)S. epidermidis–
specific TCR-transgenic CD8+T cells (BowieTg)
were adoptively transferred to wild-type mice
before colonization withS. epidermidis.(A)
Representative contour plots of IL-17A and
IFN-gproduction potential; (B) expression of
tissue residency markers CD69 and CD103 by
indicated CD8+T cell populations; (C) repre-
sentative confocal imaging volume projected
along thezaxis of epidermal skin fromS. epidermidis–colonized mice. (DandE)
Conjoined pairs ofS. epidermidis–colonized CD45.1 and CD45.2 mice were
analyzed 90 days after parabiosis surgery for cellular origin and phenotype.
(D) Frequency of host-derived f-MIIINA:H2-M3+CD8+T cells in indicated
tissues; SLN, skin-draining lymph nodes. (E) Representative contour plot of
CD69 and CD103 expression by skin f-MIIINA:H2-M3+CD8+T cells. (F)
Representative contour plot of RORgtandT-betexpressionbyCD8+T cells
from the skin ofS. epidermidis–colonized wild-type mice. (G) Representative
contour plots of RORgt, CCR6, and IL-17A expression by CD8+Tcells from the
skin ofS. epidermidis–colonized wild-type mice. (HandI)S. epidermidis–

colonized wild-type mice were exposed to bites from sand flies (L. longipalpis)

or injected intradermally (i.d.) with PBS or chitin. (H) Frequencies of TC 1

and TC17 cells with cytokine-producing potential from the skin ofS.

epidermidis–colonized wild-type mice after skin injury. (I) Representative

contour plots of IL-5 and IL-13 production potential by CD8+T cells from

the skin ofS. epidermidis–colonized wild-type mice after skin injury.

Numbers in representative plots indicate means ± SD. Bar graphs show

means ± SD. Data represent at least two experiments with four to six mice

per group. **P< 0.01 (one-way ANOVA with Holm-Šidák multiple-

comparison test).

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