Symbiotic microbiota and the development

of RORgt-expressing cells

Lymphoid Tissue Development Unit

Gérard Eberl

Self

~1012 cells ~ 104 genes The mammalian metabolome

The mammalian metabolome +

- Processing of nutrients

- Degradation of xenobiotics

- Protection from new microbes

- Regulation of epithelial homeostasis

The Superorganism

Optimal Fitness

Commensal

microbes

~1014 Bacteria

+ Archea

+ Fungi

+ Viruses

> 106 genes The microbial metabolome

+

The Host-Microbiota Superorganism

Eberl, Mucosal Immunology 2010

Bacterial Flora

Weak Immunity

Equilibrium between the microbiota

and the host

Bacterial Penetration

Systemic infection

Strong Immunity

Bacterial Penetration

Inflammation I BD

The host-microbiota equilibrium: robust homeostasis

The host-microbiota equilibrium: robust homeostasis

The tool of the host: the immune system

Mutualist Pathogen

Regulation Inflammation

MAMPs Ags

IL-2

IL-10

TGFb

...

IL-12

IL-23

TNF

…

Microbes

Immune

responses

Development of adaptive immunity

Lymphoid tissues

Lymphocyte subsets

Lymphocyte memory

G. Eberl, Mucosal Immunol 2010

The immune system to maintain homeostasis

RORgt+ lymphoid cells

Fetus Newborn / Suckling Adult

LTi cells

NK22 cells

Th17 cells

Tg cells

iNKT cells

LNs, PPs Cryptopatches ILFs

Symbiotic microbiota

IL-22 Epithelial cells

IL-17, IL-22 Epithelial cells

IL-17 Neutrophils

+

Anti-microbial peptides

Y Y Y

Y

Y Y

Y Y

Y

IgA-producing B cells

Anti-microbial peptides

Neutrophils G. Eberl, Immunological Reviews, 2011

Symbiotic bacteria of the human body

Clostridium

Bacillus

Lactobacillus

Enterococcus

(Listeria)

(Staphylococcus)

Bacteroides

Mycobacterium

Streptomyces

Bifidobacteriales

Escherichia

(Salmonella)

(Vibrio)

(Helicobacter)

(Bordetella)

(Neisseria)

Adaptive and Innate lymphoid cells

T cells

Th1

Th2

Treg

Th17

Th22

ILCs

ILC1

ILC2

ILCreg

ILC3

ILC3

NK cells

Nuocytes

?

LTi cells

LTi cells

NK22 cells

IFNg

IL-4,5,6,13

TGFb, IL-10

IL-35

IL-17

IL-22

Cherrier et al., Curr. Opin. Immunol., 2012 bis

RORgt

RORgt+ cells in the fetus: Lymphoid Tissue (LTi) cells

Cervical Mesenteric Cervical Mesenteric

Cervical Mesenteric Cervical Parathymic

RORgt ICAM-1 RORgt VCAM-1

E16.5

Eberl et al, Nat Immunol 2004

CP iILF mILF

RORgt B220 CD11c Bouskra et al, Nature 2008

Eberl et al, Science 2004

RORgt

RORgt+ cells in the adult intestine: cryptopatch LTi cells

Type 3 (RORgt+) ILCs

develop independently of microbiota

102

103

104

105

106

CD4+T

GFP+ LTi4 NK22

Nb

of

LP

L

SPF

GF

GFP+CD3-

Total Total

S. Sawa, Science 2010

days after birth

0

20

40

60

80

100

-28 -14 0 14 28 42 56

% i

n G

FP

+C

D3

-

C

D

NK22

LTi0

LTi4

2 wk

8 wk

IL-22

0 10 2 10 3 10 4 10 5

0

20

40

60

80

100

74

0 10 2 10 3 10 4 10 5

0

20

40

60

80

100

47

0 10 2 10 3 10 4 10 5

0

20

40

60

80

100

38

0 10 2 10 3 10 4 10 5

0

20

40

60

80

100

29

LTi4 NKp46+ Il22

1

10

102

103

Rela

tiv

e g

en

e e

xp

ressio

ns

LT

i 4

LT

i 4

E15 2 wk 8 wk

LT

i 4

NK

p4

6+

NK

p46

+

*

*

Il-22 production by RORgt+ ILCs decreases with age

S. Sawa, Nat Immunol, 2011

2 wk

0

1

2

3 43 3.6

LTi4 NKp46+

0 10 2

10 3

10 4

10 5

0 10 2

10 3

10 4

10 5

0 10 2

10 3

10 4

10 5

0

0.1

0.2

0.3

0.4

0

0 10 2

10 3

10 4

10 5

0

0.3

0.6

0.9

1.2

0

IL-17A

8 wk

Il17a

Rela

tiv

e g

en

e e

xp

ressio

ns

1

10

102

LT

i 4

LT

i 4

E15 2 wk 8 wk

LT

i 4

NK

p46

+

NK

p46

+

N.D

N.D

N.D

Il-17 production by RORgt+ ILCs decreases with age

S. Sawa, Nat Immunol, 2011

Rela

tiv

e g

en

e e

xp

ressio

ns

Il22

SPF GF

RORgt+ILCs CD4+T

- + - +

1

NK

p46

+

LT

i 4

NK

p46

+

10

102

103

*

*

N.D

N.D

LT

i 4

SPF GF

*

*

NKp46+

LTi4

GF SPF

0 10 2 10 3 10 4 10 5

0

20

40

60

80

100

7

0 10 2 10 3 10 4 10 5

0

20

40

60

80

100

27

0 10 2 10 3 10 4 10 5

0

20

40

60

80

100

15

0

20

40

60

80

100

0 10 2 10 3 10 4 10 5

38

IL-22

Il-22 production by RORgt+ ILCs is repressed by microbiota

S. Sawa, Nat Immunol, 2011

Development and function of RORgt+ innate lymphoid cells

days after birth -14 0 14 28 42 56

ckitlow

NK22

LTi0

LTi4

Lymph nodes

Peyer’s patches

Cryptopatches

ILFs

Epithelial

defense

Microbial

colonization

Clustered LTi4 and LTi0 cells

Effector P46 and Ckitlow cells

Pre-empt microbes

Epithelial cells IL-25 CPs/ILFs/B cells

IL-17/22

The function of innate lymphoid cells

Cherrier et al., Curr. Opin. Immunol., 2012 bis

CLP

IL-7R+ IL-7

HSC

4b7-

RORgtlo

4b7+

RORgt- NK Nuocyte

LTi NK22

Notch

Id2

Id2

Tox

RORgt

Id2

RORgt

Notch

AhR

B

T

Notch

IL-12

IL-15

IL-18

IL-25

IL-33 RORgt

AhR

IL-23

ROR Runx1

Tox

The development of ILCs

Cherrier et al., J. Exp.Med. 2012

RORgt+ lymphoid cells

Fetus Newborn / Suckling Adult

LTi cells

NK22 cells

Th17 cells

Tregs

Tg cells

iNKT cells

LNs, PPs Cryptopatches ILFs

Symbiotic microbiota

IL-22 Epithelial cells

IL-17, IL-22 Epithelial cells

IL-17 Neutrophils

+

Anti-microbial peptides

Y Y Y

Y

Y Y

Y Y

Y

IgA-producing B cells

Anti-microbial peptides

Neutrophils G. Eberl, Immunological Reviews, 2011

Lymphoid Tissue Development Unit

Sponsors

Excellence Team

Shinichiro SAWA Marie CHERRIER

Caspar OHNMACHT

Gnotobiology Platform Marion Bérard

Christophe Brézillon

José Perez

Paroi bactérienne Ivo Gomperts-Boneca

Catherine Werts

Immunité Innée Jim Di Santo

Naoko Satoh-Takayama

CIGM Francina Langa

Nadine Cerf-Bensussan

Valérie Gaboriau-Routhiau

Daniel Cua

Mélanie Kleinschek