Immun tolerans mekanizmaları ve regülatör

T hücreleriMübeccel Akdis, MD, PhD

Swiss Institute of Allergy and Asthma ResearchSIAF Davos

Akdis

antigen

naiveT cell

differentiation signals

Th2

Th2 switch and expansion

innateimmune responsestimulatingsubstances

Th17

Th17 switch and expansion

Th1

Th1 switch and expansion

Th9

Th9 switch and expansion

differentiationfactors

effector cytokines IFN-g IL-4,IL-5, IL-13 IL-9 IL-8, IL-17A, E,

IL-22, IL-26

IL-12 IL-4 IL-4, TGF-b

TGF-b, IL-6, IL21, IL-23

induction of IgG4suppression of IgE

TReg

mastcell

Th2

basophil

Direct and indirect suppressive effects

on mast cells, basophils

and eosinophils

endothelial cells

Suppression of effector T cell

migration to tissues

eosinophil

Th1

Suppression of Th2 effector

cells

Suppression of Th1

effectorcells

Suppression ofinflammatory

dendritic cells.Induction of IL-10-

producing dendritic cells

Suppression of Th17

effectorcells

Th17

remodelling in the tissues

akdis

1. Allergen-specific immunotherapy

2. Natural high dose allergen exposure

3. Antibody regulation

cyto

kine

(ng/

ml)

days

IL-13

IL -10

3

1

2

0

4

IL-5

7 280 70

IFN-g

TGF-b *

IL-10 and TGF-b in peripheral T cell tolerance

during aeroallergen SIT

n= 10, SIT cluster protocol, Der p 1-stimulated PBMC EJI.2003

B

A

u.s.Der p 1

4

8

0

4

8

0

**

before SIT after SIT

1x1 0

3

5x1 0

3

15x1

03

1x10

3

5x1 0

3

15x 1

03

PB

MC

CD4+CD25+ CD4+CD25+

[3 H] T

dR [c

pmx1

0 -3]

PB

MC

10

20

0

10

20

0

**

[3 H] T

dR [c

pmx1

0 -3]

Der p 1u.s.

CD4+CD25-

1x10

3

5x10

3

15x 1

03

PB

MC

CD4+CD25+

CD4+CD25-

1x10

3

5x1 0

3

15x 1

03

PB

MC

CD4+CD25+

IL-4 IL-100

2.5

5

7.5

10

frequ

ency

/ 10

.000

cel

ls Allergic

IFN-g

Der p 1Bet v 1

**

*

****

***

Aeroallergen-specific T cell frequency in health and allergy

Pyr c 5Cor a 1

IL-4 IL-10

Healthy

IFN-g

J Exp Med 2004

0

20

40

60

80

100

[3 H]T

dR c

pmx1

000

+Der p 1-IL-4+

+Der p 1

+Der p 1-IL-10+

+Bet v 1-IL-10+

+Bet v 1-IL-4+

+Bet v 1

+Bet v 1-IL-10+

+Der p 1-IL-10+

+ + +

+

+

+

+

+ +

+ + +

+

+

+

+

+ +

* *

TReg cells in allergy: a question of antigen specificity

J Exp Med 2004

Beekeepers show way to allergen tolerance

The Journal of Experimental Medicine, November 10, 2008• Nicole LeBrasseur

Nature Reviews Immunology

Beekeepers hold clues for T-cell tolerance By Sarah Allan, PhD

New Scientist MagazineA sting a day keeps the allergy away

18 November 2008 by Ewen Callaway

Antigen-specific peripheral T cell tolerance after live bee

stings

bee keepingseason

0

20

40

60

80

[3 H]-t

hym

idin

e in

corp

orat

ion

(cpm

x 1

000)

J F M A M Ju Jl A S O N Dmonths of the year

PLA

02468

101214

J F M A M Ju Jl A S O N Dmonths of the year

bee

stin

gs /

mon

th

J. Exp. Med., 2008

0

10

20

30firs

t be

e stin

gaft

er 7

daft

er 1

mfirs

t be

e stin

g

next

year

late phase swellinglesion size 2: > 3 cm1: < 3 cm0: no lesion

pain and immediate wheal and flare reactiondo not change

bee keepingseason

0

20

40

60

80

J F M A M Ju Jl A S O N D

[3 H]-t

hym

idin

e in

corp

orat

ion

(cpm

x 1

000)

months of the year

lesi

on s

ize

5 beekeepers three years follow up

Frequency of PLA-specific cytokine secreting T cells before and after live bee stings

*: p<0.001n:6 beekeepers

Decreased frequency of PLA-specific IL-4- and IFN-g-secreting T cells,increased frequency of IL-10-secreting T cells is observed 7 days after ≥20 bee stings

0

5

10

15

IL-4 IFN-g IL-10

before7 d after

*

*

*

spec

ific

CD

4+ T c

ells

frequ

ency

/ 10

4 cel

ls

Th2

mastcell

histamine

HR2 suppresses APC capacity, increases IL-10 and suppresses IL-12 in

maturing dendritic cells

HR2 on Th2 cellsuppresses IL-4, IL-13

and proliferation

HR2

IL-1

0

immature DC

mature DC

IL-10

HR2

Th1

HR1 on Th1 cellenhances IFN-g and proliferation,

increases effector functions and tissue injury

TReg

histamine

HR1

enhancedsuppressivecapacity of Treg cells

PLA

024

14

b a

HR1

PPD

0.51.01.5

b a0

PHA

b a

05

101520

b a

HR2

05

1015

b a05

1015

05

1015

00.51.01.52.0

HR4

b a

b a b a00.51.01.52.0

b a01.02.03.0

FoxP30

1.02.03.0

b a01.02.03.0

b a b a01.02.03.0

rela

tive

mR

NA

expr

essi

onA B

00.20.40.60.81.01.21.4

0 -6 -5 -4 0123456

0 -6 -5 -40

1

2

3

0 -6 -5 -4

IL-4 IL-13 IL-10

histamine+ranitidine

histamine [M] log10

cyto

kine

(ng/

ml)C

FoxP3

IL-4

IFN

-gIL

-10

IL-4

IFN

-gIL

-100

204060

HR2

0

5

10

15

rela

tive

mR

NA

ex

pres

sion

before after 7 d

CD

4+

CD

25+

CD

4+

CD

25-

CD

4+

CD

25+

CD

4+

CD

25-

HR2 is upregulated in specific T cells after multiple beestings and induces IL-10 production

J. Exp. Med., 2008

non-inflammatory Ab isotypes

IgAIgG4

inflammatory Ab isotypes

IgG1, IgG2, IgG3

IgM IgE

?

??

Th1

Th 17

Th2

Treg

PLA-specific IgE:IgG4 antibody ratios

anti-

PLA

IgE

: Ig

G4

[log1

0]

allergic SIT bee keeper

-5

-4

-3

-2

-1

0

IL-10 suppresses IgE

and induces IgG4

IgG4 is non inflammatory Does not exist in miceReflects dose of exposure and accompanies all high dose antigen exposure models, bee keepers, helminth infections (in the absence of high TLR triggers) Does not activate complementNo specific Fc receptors for IgG4 are defined so farIn vitro studies demonstrate nonanaphlactogenic activityby blocking of IgE binding and dual specificityMolar quantities are at least 1000 X higher than IgE

Allergy, 2008

Allergy, 2008

J Clin Invest 1996J Clin Invest 1998FASEB 1999FASEB 2001Immunology TodayImmunology 2001Nature 2001Eur J Imm 2003FASEB 2003Nature Rev Imm 2003J Exp Med 2004JACI 2005Nature Rev Imm 2006Curr Opin Imm 2006JACI 2007-2008JEM 2008

allergen-specificTreg cell

multiplesuppressor

factorsIL-10,TGF-b,IL-10R,TGF-bR,CTLA-4, PD-1, HR2

induction of IgG4, IgAsuppression of IgE

IL-10TGF-b

Immune tolerance to allergens in healthy immune response and successful SIT

eosinophil

IL-10TGF-b

decreased TH2 cytokines

suppression of effector cells

• Beate Rückert• Sven Klunker• Flurina Meiler• Willem Van De

Veen• Judith Zumkehr• Mario Ziegler• Tunc Akkoc• Umut

Kücüksezer• Günnur Deniz

• Cezmi A. Akdis• Reto Crameri• Kurt Blaser

• U. Müller, Bern• B. Wüthrich, Zurich• M. Jutel, Wroclaw,

SIAF• R. Valenta, Vienna• E. Jensen-Jarolim,

Vienna• M. van Hage-Hamsten,

Stockholm• G. Gavfelin,

Stockholm• R. Kroczek, Berlin• M. Colonna, St Louis• E. Flory, Langen• S. Viehts, Langen• H. Fiebig, Reinbek• C. Heusser, Basel• E. Hamelmann, Berlin• M. Larchè, London• A. Verhoef, London• S. Alkan,

Minneapolis• R. Lauener, Zurich• B. Ballmer-Weber,

Zurich

• R. Disch• B. Wüthrich• P. Schmid-

Grendelmeier• W. Kneist• M. Schliz• D. Kleeman• W. Deglmann• H. Behrendt, • C. Traidl-

Hoffmann, Munich

Immune tolerance Mechanisms of atopy

• T. Watanabe, Osaka

• R. Koga, Fukuoka• Marek Jutel

Immune regulation by histamine

World Immune RegulationDavos29 March- 1 April 2009

1

1. Basophil, mast cell desensitisation, early events related to APC (DC, B cell),effect of adjuvants

3. Regulation of B cells, antibodies and related phenomena

32

2. Regulation of T cells and related phenomena

hoursdays weeks months years

mechanisms of SIT

start

day 1

day 7

1 mon

th

6 mon

ths 1-3 years

rela

tive

chan

geEarly decrease in mast cell and basophil activityfor degranulationand systemic anaphylaxis

T cell tolerance induction of IL-10 and sometimes TGF-b secreting Tr1 cellssuppression of Th2-Th1 cells

Mechanisms of allergen-specific immunotherapy

decrease in tissue mast cells and eosinophilsand decreased release of their mediators

start

day 1

day 7

1 mon

th

6 mon

ths 1-3 years

rela

tive

chan

ge

Type I skin test reactivity

Mechanisms of allergen-specific immunotherapy

specific IgE

specific IgG4

Stim

ulat

ion

Inde

x

0

5

10

15

20

before day 7 day 28

Stim

ulat

ion

Inde

x

IL-10 neutralizing

0

5

10

15

20

25

before day 7 day 28

IL-13IL-10IL-5

IL-4IFN- g3H-Thymidine incorporation

IL-10-induced peripheral T cell tolerancein bee venom specific-immunotherapy

J. Clin. Invest.1996; 98: 1676J. Clin. Invest.1998; 102: 98

day 28day 7day 0

IL-10 production in T cells during specific immunotherapy

100 103

CD4+ CD25+ T cells

even

ts

J. Clin. Invest.1998; 102: 98

IL-10

even

ts

10 0 10

PLA stimulated T cells

3

IL-10

Direct demonstration

TcRb240 - 285 bp

specimen control size ladder

before afterP

TcRb 240 - 285 bp

TcRb 285 - 325 bp

TCRb 170 - 210 bp

IFN-g IL-4IL-10 IFN-g IL-4 IL-10 N

PLA-specific IL-4-, IFN-g- and IL-10-secretingT cells are expanded and their clonality is determined

Same T cells skew to Tr1 cells after bee stings

J. Exp. Med., in press

TcR vb12

0.0

2.5

5.0

7.5

10.0

12.5

b a b a b a

TcR vb2

0

10

20

30

b a b aIL-4 IL-10 IFN-g IL-4 IL-10

Same T cells skew to Tr1 cellsafter bee stings

PLA-specific IL-4-, IFN-g- and IL-10-secretingT cells are expanded and TcR vb are determined

J. Exp. Med.2008

0

2

4

6

8

10

0 0.1 0.5

+sTGF-bR

+anti-IL-10R

PBMC before SIT+CD4+CD25 after SIT

PBMC before SIT

+anti-IL-10R +sTGF-bR

C[3 H

] TdR

[cpm

x10

-3]

Der p 1 [µM]

*

*

**

(IL-1

0/ IF

N-g

ng/

ml)

0

200

400

600

IL-13 IFN-g IL-10

(IL-4

/IL-1

3 pg

/ml)

IL-40

2

4

6

* * **

7d afterstings

beforestingsJan/Feb

A rapid in vivo switch to specific IL-10-producing T cells