June 10-12th ,2013

Veyrier du Lak, France

Targeting Commensal Flora to better shape

protective immune responses for better disease

prevention and therapy

Hiroshi Kiyono Division of Mucosal Immunology

International Research and Development Center for Mucosal Vaccine The Institute of Medical Science

The University of Tokyo

Mucosal Connections between

Immune Regulation and Commensal Microbiota

Targeting Commensal Flora to Better Shape Protective Immune Responses for

Better Disease Prevention and Therapy

Thank You for Your Kind Invitation

J. Numazu

M. Igeta

Merci Pour Votre Invitaion de Denre

Prof. Nicolas Burdin Prof. Gerard Eberl Prof. Patricia Londono-Hayes Prof. Jacques Louis

Hydrangea

Cherry Blossoms

TLR5+ CD11cH

CD11bH cells

TLR5

Th1

Retinoic Acid(RA)

ATP

Bacterial ATP

Treg CD70H CD11cL

CD11b+ cells

ATP Receptor

IgA Th17 Mast cells

Peyer’s patch

Treg

Ba

Th1

DCs

Th2

Th2

M Cells

Ba

Ba Bm

Th3

TipDCs

Tfh

RA

SFB

Th9

TMP-IMSUT

LTi

IL-22 IL-33

The Mucosal Immune System Pathogenic and Commensal Bacteria

Inductive Site

Effector Site

Microflora: 100 Trillion Organisms with 300 Spices

DC

Mφ

M Cell

Nutrients Allergens Bacteria &

Virus

GP-2 UEA-1 NKM16-2-4 Spi-B RANK/RANKL TLR1,2 & 4

CCR5 CXCL16

C5aR (OmpH)

Enriched with IgA Plasma Cells GALT

Peyer’s patch

Peritoneal Cavity

Ba B-2

Ba B-1

Intestinal IgA Induction System

Intestine is a Major Source of IgA: Plasma Cells are Originated from Peyer’s Patches

and Peritoneal Cavity

Critical Roles of IgA for the Mucosal Protection and Symbiosis

Induction and Maintenance of IgA Plasma Cells: Effect of Commensal Microbiota ?

Diversity and Function of IgA? TMP-IMSUT

Presence of Two Subsets of IgA Cells Based on the CD11b Expression

6.6

14.0

CD

11b

IgA

Peyer’s Patch Dependency

14.7

0.8

Peyer’s Patch Deficient

14.4

4.3

CD

11b

IgA

Wild Type

CD11b+ IgA+

CD11b- IgA+

Two Subsets of IgA Cells Based on the Expression of CD11b

2.7

13.3

Antibiotics Treatment

Reduction of CD11b+IgA+B Cells

Influence of Commensal Microflora

Intestinal Flora and Peyer’s Patch Dependent Induction of CD11b+ IgA Cells

1.4

MyD88 KO

J. Kunisawa, et al., Nat. Commun. (2013)

NIBIO

Two Subsets of IgA Plasma Cells

Ba

Oral Antigens

Bm

Class Switch

CD11b+

CD11b-

High Proliferation High IgA Secretion

Commensal Flora・MyD88- and PP- Dependency

Importance of Peyer’s Patch- & Commensal Flora- dependent CD11b+IgA Plasma Cells for the Intestinal Immunity

GALT Peyer’s patch

TMP-IMSUT

Commensal Mirobiota

CD11b Expressions Specificity Function (?)

Symbiosis

Pathogens

Protection

Stomach

Cecum

Large Intestine

Small Intestine

Gut-associated Lymphoid Tissues as the Intra-tissue Cohabitation Niche for Commensal Bacteria in

the Control of Mucosal Homeostatic Condition

DCs B Cells T Cells

Peyer’s patch

Presence of Alcaligenes in Mouse, Primate and Human PPs

Alcaligenes-specific IgA

Possible Contribution of IgA for Intra-tissue Co-habitation

Analysis of Indigenous Bacteria in PP

Alcaligenes Ochrobactrum

Serratia Burkholderia Other bacteria

72%

10% 4%

6%

10%

OD 4

50

Normal IBD (CD)

TMP-IMSUT Obata et. al., Proc. Natl. Acad. Sci. 107: 7419-7424, 2010

72%

Anti - Alcaligenes Fecal IgA

SPF

GF

＊ ＊

＊ ＊

＊ ＊

＊ ＊

3 4 5 6 7 8 0.0

0.2

0.4

0.6

0.8

Dilution (log 2 )

OD

450

SPF

GF

＊ ＊

＊ ＊

＊ ＊

＊ ＊

3 4 5 6 7 8 0.0

0.2

0.4

0.6

0.8

Dilution (log 2 )

OD

450

＊ ＊

＊ ＊

＊ ＊

＊ ＊

3 4 5 6 7 8 0.0

0.2

0.4

0.6

0.8

Dilution (log 2 )

OD

450

Anti - Alcaligenes Serum IgG

SPF

GF

1 2 3 4 5 6 7 0.0

0.1

0.2

Dilution (log 2 )

OD

450

SPF

GF

1 2 3 4 5 6 7 0.0

0.1

0.2

Dilution (log 2 )

OD

450

SPF

GF

0.0

0.2

0.4

0.6

0.8

0.0

0.1

0.2

Dilution (log 2 )

3 4 5 6 7 8

Dilution (log 2 )

1 2 3 4 5 6 7

OD

45

0

OD

45

0 SPF

GF

＊ ＊

＊ ＊

＊ ＊ ＊ ＊

Anti - Alcaligenes

SPF

GF

＊ ＊

＊ ＊

＊ ＊

＊ ＊

3 4 5 6 7 8 0.0

0.2

0.4

0.6

0.8

Dilution (log 2 )

OD

450

SPF

GF

＊ ＊

＊ ＊

＊ ＊

＊ ＊

3 4 5 6 7 8 0.0

0.2

0.4

0.6

0.8

Dilution (log 2 )

OD

450

＊ ＊

＊ ＊

＊ ＊

＊ ＊

3 4 5 6 7 8 0.0

0.2

0.4

0.6

0.8

Dilution (log 2 )

OD

450

Anti - Alcaligenes

SPF

GF

1 2 3 4 5 6 7 0.0

0.1

0.2

Dilution (log 2 )

OD

450

SPF

GF

1 2 3 4 5 6 7 0.0

0.1

0.2

Dilution (log 2 )

OD

450

SPF

GF

0.0

0.2

0.4

0.6

0.8

0.0

0.1

0.2

Dilution (log 2 )

3 4 5 6 7 8

Dilution (log 2 )

1 2 3 4 5 6 7

OD

45

0

OD

45

0 SPF

GF

＊ ＊

＊ ＊

＊ ＊ ＊ ＊

Goto Y. and Kiyono H. Immunol. Rev. 245:147-63,2012

5 μm 20 μm

Alcaligenes GP2 DAPI

M Cell

Transcytosis of PP-resident Bacteria, Alcaligenes by M Cells

TMP-IMSUT

GFP-Alcaligenes Intestinal Loop Assay

5 Hours

Analysis

Sato, S. et al. Mucosal Immunol. Epub ahead of print, 2012.

DC

Critical Role of Innate Lymphoid Cells for the Control of Intra-tissue Cohabitated Commensal Bacteria in

Gut-associated Lymphoid Tissues

TMP-IMSUT

Ba Th

Alcaligenes

Regulation of Homeostasis between Intra-tissue Bacteria and Mucosal Immunity

Ba

ILC

Obata et. al., Proc. Natl. Acad. Sci. 107:7491-7424, 2010 Ｓｏｎｎｅｎｂｅｒｇ et. al., Ｓｃｉｅｎｃｅ 336：１３２１-１３２５、 201２

ILC TFH

TReg ILC

CD127+ CD45INT RORgt+ IL-22+ CD90.2+

TNFa ILC

_ +

Commensal Bacteria (Alcaligenes)

10５ 102 100 0

(CFU/g)

(pg/ml)

90 0

_

+

(Weight: g)

Spleen

DC

The Mucosal Immune System for the Creation of Immunological Co-habitation Niche for

Intra-tissue Commensal Bacteria

TMP-IMSUT

SIgA

Ba T

Induction & Regulation of

IgA Alcaligenes

Ba

Ba

ILC

ILC

ILC

Calprotectine

IL-22

Inflammation

Breakdown of ILC System

Obata et. al., Proc. Natl. Acad. Sci. 107:7491-7424, 2010 Ｓｏｎｎｅｎｂｅｒｇ et. al., Ｓｃｉｅｎｃｅ 336：１３２１-１３２５、 201２

ILC for Immunological Balance with Intra-tissue Commensal Bacteria

IgA

Peyer’s patch

Treg

Ba

Th1

DCs

Th2

M Cells

Ba

Ba Bm Th3

TipDCs

Tfh

RA

SFB

TMP-IMSUT

IL-22

IL-33

Commensal and Pathogenic Bacteria

Inductive Site Effector Site

ILC ILC

ILC

ILC

CD11b+

CD11b-

ILC

ILC

Fut2 Fut2

Alcaligenes Intra-tissue Habitation

Mucosal Connections for Integrated Regulation among Innate –Acquired Immune Cells , Epithelial Cells and Commensal Microbiota

University of Tokyo (IMSUT) Hiroshi Sagara

Stanford University Anson W. Lowe (GP2-/-Mice)

Acknowledgement

University of Michigan Steven E. Domino (Fut1lacZ/lacZ Mice)

Kitazato Univ. Hidenori Matsui

Riken Yoshimi Benno

Mitsuo Sakamoto

Yakult Central Institute Yoshinori Umesaki

Takahiro Matsuki

Hiromi Setoyama

Columbiana University Ivaylo I. Ivanov

University of Pennsylvania Gregory F. Sonnenberg David Artis M. Celeste Simon (Spib-/-Mice)

Kyushu University

Masatoshi Goto

Hideki Iijima

Akira Mukai

Shinichiro Shinzaki

Osaka University

Understanding of Mucosal Immunity for Vaccine Development

Tokyo Mucosal Patches (TMP)

2013

2009-2010

M. Igeta

Jun Kunisawa Shintaro Sato Yoshiyuki Goto

Naoko Shibata Satoshi Kaneto Yoshikazu Yuki

Yuko Takahashi Hideaki Okura Takashi Obata

Makuhari Messe

The 42nd Annual Meeting of Japanese Society for Immunology

National, but Open to All Over the World

December 11th-13th 2013

Immunology Bring US Together

We Welcome All of You

Joint Symposium with SFI, AAI, DGFI, CSI, KAI and SIgN