Download IL-31

Dysregulation of innate and adaptive
immunity in patients with atopic dermatitis:
Impact of IL-31/IL-31R & staphylococcal
α-toxin
Sadaf Kasraie, Ph.D.
Division of Immunodermatology and Allergy Research,
Department of Dermatology and Allergy
Activation
IL-31/IL-31R
CD 4+ T- Cells
Induces pro-inflammatory cytokines in some human
cell lines so far 1,2
(24 KDa)
Receptor 1: IL-31RA (GPL) (120 KDa)
Receptor 2: OSMR (180 KDa)
Epithelial cells
Keratinocytes
Jak/STAT, PI3 kinase/Akt and MAP kinase signalling pathway
Sadaf Kasraie
Division of Immunodermatology and Allergy Research
Activated
monocytes and
macrophages
Eosinophils
1 Dambacher J. et al. 2007. Gut 56:257-65.
Modified from Le
Saux S. et al. J Biol Chem. 2010. 285:3470-7
2 Yagi Y. et al. 2007. Int J Mol Med 19:941-6.
Role of IL-31
IL-31 overexpression may lead to :
• Pruritus
• Alopecia
• Skin lesions
• Airway hypersensitivity
• Atopic dermatitis (atopic eczema)
• Allergy
Sadaf Kasraie
Division of Immunodermatology and Allergy Research
Review
• Transgenic overexpression of IL-31 in lymphocytes induces severe
pruritus in mice (Dillon et al., Nat Immunol 2004).
• IL-31 is overexpressed in the skin of patients with atopic dermatitis
(Sonkoly et al., JACI 2006).
• Serum IL-31 levels are significantly higher in patients with AD and
correlated with disease activity in AD (Raap et al., JACI 2008).
• Staphylococcal superantigens rapidly induce IL-31 expression in
atopic individuals in vivo and in leukocytes in vitro (Sonkoly et al.,
JACI 2006).
Sadaf Kasraie
Division of Immunodermatology and Allergy Research
Approach
To investigate
• the expression,
• the regulation of IL-31R,
• functional effects of IL-31 in human primary
cells (monocytes, macrophages, keratinocytes
and eosinophils): Regulation of STAT and
MAPK signalling pathways as well as cytokine
secretion
Sadaf Kasraie
Division of Immunodermatology and Allergy Research
Expression of IL-31RA on Immune Cells & Keratinocytes
Non Stimulated
Stimulated*
Monocytes
-
++1,2
Macrophages
-
++1,2
DC: moDC
-
-
DC: IDEC
-
-
Lymphocytes
-
-
Keratinocytes
+3
++
Eosinophils
+4
+
1
Kasraie S. et al. 2010. Allergy. 65:712–721.
2 Kasraie S. et al. 2011. Allergy. 66:845-52.
3 Kasraie S. et al. 2013. Allergy. 68:739-47.
4 Raap U, Gehring M, Kasraie S. et al. Submitted.
Sadaf Kasraie
Division of Immunodermatology and Allergy Research
* SEB, α-toxin or TLR-2 agonists
Macrophages
Kasraie and Werfel. 2013. Mediators Inflamm 2013:942375
Modified from Werfel T. 2009. J Invest Dermatol 129:1878-91
Sadaf Kasraie
Division of Immunodermatology and Allergy Research
Staphylococcal exotoxins significantly up-regulate IL-31RA
expression in human monocytes and macrophages at the
mRNA level
A) Monocytes
Sadaf Kasraie
Division of Immunodermatology and Allergy Research
B) Macrophages
Staphylococcal exotoxins
exotoxins up-regulate
up-regulate IL-31RA
IL-31RA
Staphylococcal
expression
in human
monocytes
and
at the
expression
in human
monocytes
atmacrophages
the protein level
protein level
A) Monocytes
Sadaf Kasraie
Division of Immunodermatology and Allergy Research
B) Macrophages
Signalling pathways of IL-31 in human macrophages
pSTAT-1
β-actin
pSTAT-5
β-actin
pERK1/2
β-actin
Sadaf Kasraie
Division of Immunodermatology and Allergy Research
Summary of IL-31 effects upon staphylococcal
exotoxins stimulation in monocytes & macrophages
Cells
Cytokine
MΦ & PBMCs
IL-6
MΦ & PBMCs
IL-1β
MΦ
IL-18
MΦ
IL-8
MΦ
IL-10
MΦ
TNF-α
MΦ
MIP-1α
MΦ
MCP-1α
MΦ
IL-12p40
MΦ
IL-12p70
MΦ
IL-23p19/p40
Sadaf Kasraie
Division of Immunodermatology and Allergy Research
SEB
α-toxin
IL-31 induces pro-inflammatory cytokines in human
macrophages following up-regulation of the IL-31R with
staphylococcal exotoxins
Sadaf Kasraie
Division of Immunodermatology and Allergy Research
IL-31 down-regulates IL-12 in human macrophages
following up-regulation of the IL-31R with staphylococcal
exotoxins
Sadaf Kasraie
Division of Immunodermatology and Allergy Research
Keratinocytes
Kasraie and Werfel. 2013. Mediators Inflamm 2013:942375
Modified from Werfel T. 2009. J Invest Dermatol 129:1878-91
Sadaf Kasraie
Division of Immunodermatology and Allergy Research
TLR-2 ligand Pam3Cys up-regulates IL-31RA expression
in human primary keratinocytes at the mRNA level
*
*Pam3Cys-SK4: N-Palmitoyl-S-[2,3-bis (palmitoyloxy)-(2RS)-propyl]-[R]-cysteinyl-[S]-seryl-[S]-lysyl-[S]-lysyl-[S]-lysyl-[S]-lysine
Sadaf Kasraie
Division of Immunodermatology and Allergy Research
Pam3Cys or IFN-γ up-regulates IL-31R expression on
human primary keratinocytes at the protein level
NS= Not Stimulated
Sadaf Kasraie
Division of Immunodermatology and Allergy Research
Signalling pathway of IL-31 in human keratinocytes
pSTAT-3
STAT-3
Sadaf Kasraie
Division of Immunodermatology and Allergy Research
Summary of IL-31 effects upon Pam3Cys or IFN-γ
stimulation in keratinocytes
HPKs
Chemokine/Cytokine
Pam3cys
IFN-γ
Foreskin &
Hair
CCL2
+
+
Foreskin
VEGF
+
-
Foreskin
CCL22
+
-
Foreskin
CCL20
-
-
Foreskin
EGF
Foreskin
MMP-9
Foreskin &
Hair
IL-1β
-
-
Foreskin &
Hair
IL-6
-
-
Foreskin &
Hair
IL-8
-
-
Sadaf Kasraie
Division of Immunodermatology and Allergy Research
IL-31 induces CCL2 secretion in human primary
keratinocytes following up-regulation of the IL-31R with
Pam3Cys or IFN-γ
Sadaf Kasraie
Division of Immunodermatology and Allergy Research
IL-31
Staphylococcal exotoxins
TLR-2 ligands or IFN-γ
Activated
monocytes and
macrophages
STAT-1/5,
ERK1/2
pro-inflammatory cytokines
(IL-1β, IL-6, IL-18)
Sadaf Kasraie
Division of Immunodermatology and Allergy Research
Eosinophils
Keratinocytes
STAT-3
CCL2, VEGF
STAT-3,
ERK1/2
CCL26, release of
superoxide species
Atopic dermatitis (AD) vs healthy controls (HC)
Patient 1 with AD
Sadaf Kasraie
Division of Immunodermatology and Allergy Research
Patient 2 with AD
IFN-γ
leads
to significantly
higher
of IL-31RA
IL-31
induces
greater levels
of expression
IL-1β secretion
in
on monocytes
from
patients
compared
to
monocytes
from
patients
withwith
AD AD
compared
to HC
healthy controls (HC)
Sadaf Kasraie
Division of Immunodermatology and Allergy Research
Pam3Cys
up-regulates
expression
IL-31
induced
lower levelsIL-31RA
of CCL2
secretion in
keratinocytes
bothwith
fromAD
ADcompared
and HC to HC
keratinocytes
from patients
Sadaf Kasraie
Division of Immunodermatology and Allergy Research
Impaired TLR-2 expression in keratinocytes from
inflamed AD skin compared with healthy controls
Sadaf Kasraie
Division of Immunodermatology and Allergy Research
Conclusion
IL-31
IL-31
Homey et al. 2006. J Allergy Clin Immunol. Review.
•
•
•
•
The Th2 cytokine IL-31 induces itch.
IL-31 has inflammatory properties.
The IL-31R is expressed on immune cells and keratinocytes.
The IL-31R is regulated by staphylococcal-derived molecules.
Sadaf Kasraie
Division of Immunodermatology and Allergy Research
Conclusion & Outlook
• Evidence for a functional role of IL-31 on human eosinophils.
• Novel implications for treatment options in eosinophil associated
diseases including AD.
• Colonization with S. aureus and presence of IL-31 in the skin may
represent important trigger factors in the complex amplification
cycle of atopic skin inflammation.
• Antiseptic strategies as well as targeting IL-31 and its receptor in
atopic individuals may represent useful tools to support the
therapeutic management of patients suffering from AD.
Sadaf Kasraie
Division of Immunodermatology and Allergy Research
Acknowledgment
Ulrike Raap
Margarete Niebuhr
Kathrin Baumert
Gabriele Begemann
Maria Gschwandtner
Manuela Gehring
Jana Zeitvogel
Christina Hartmann
Hari Balaji
Susanne Mommert
Susanne Hradetzky
Thomas Werfel
Armin Braun
Wolfgang Baeumer
All members of the Dept. of Dermatology & Allergy Family!
Sadaf Kasraie
Division of Immunodermatology and Allergy Research
Acknowledgment
Ulrike Raap
Margarete Niebuhr
Kathrin Baumert
Gabriele Begemann
Maria Gschwandtner
Manuela Gehring
Jana Zeitvogel
Christina Hartmann
Hari Balaji
Susanne Mommert
Susanne Hradetzky
Thomas Werfel
Armin Braun
Wolfgang Baeumer
All members of the Dept. of Dermatology & Allergy Family!
Sadaf Kasraie
Division of Immunodermatology and Allergy Research
RE Schmidt
Susanne Kruse
Marlies Daniel
Sadaf Kasraie
Division of Immunodermatology and Allergy Research
HBRS et al. 2010. MHH info.
Overview
Atopic Dermatitis (atopic eczema) is a
chronically relapsing inflammatory skin
disease
Prevelance: 10 to 20% in children and 1% to
3% in adults
Causes: Although it is such a common
disease, the immunopathogenesis is not
completely understood
Sadaf Kasraie
Division of Immunodermatology and Allergy Research
Trigger factors in Atopic Dermatitis
Food allergens
Hormones & related
Factors
Autoantigens
Irritable
substances
Skin colonizing
microorganisms
Genetic
background
Inhalant allergens
Environment
Psychological stress
Werfel et
Sadaf Kasraie
Division of Immunodermatology and Allergy Research
Werfel et
al. Leitline Atopisches Ekzem www.awmf.de 2002 (Revision 2007)
al. HTA Bericht Neurodermitis www.egms.de 2006
IL-31 induces IL-12 suppression via ERK 1/2
phosphorylation in human macrophages following
up-regulation of the IL-31R with staphylococcal exotoxins
Sadaf Kasraie
Division of Immunodermatology and Allergy Research
Eosinophils
Modified from Werfel T. 2009. J Invest Dermatol 129:1878-91
Sadaf Kasraie
Division of Immunodermatology and Allergy Research
Human eosinophils express the IL-31RA
A) At the mRNA level
Sadaf Kasraie
Division of Immunodermatology and Allergy Research
B) At the protein level
Signalling pathways of IL-31 in human eosinophils
pSTAT-3
pERK1/2
STAT-3
Sadaf Kasraie
Division of Immunodermatology and Allergy Research
ERK1/2
Human eosinophils are activated by IL-31 with release of
CCL26 and reactive oxygen species
Sadaf Kasraie
Division of Immunodermatology and Allergy Research
Functional effect of IL-31 in eosinophils
220,00
4
3.0
***
200,00
180,00
*
**
**
1.5
**
0.0
Fluorescence intensity
Chemotactic Index
12
Medium
160,00
IL31 100ng
IL31 100ng + Block (aIL31RA)
140,00
C5a
120,00
100,00
M Co 1 10 50 100
IL-31
nAb 1 10 50 100
IL-31 + nAb
80,00
1
4
7
10 13 16
19 22 25 28
31 34 37
Tim e (seconds)
2 Raap
U, Gehring M, Kasraie S. et al. Submitted.
Sadaf Kasraie
Division of Immunodermatology and Allergy Research
40 43 46 49
Th2
Th1
Th1
Th1
Th1 Th1
MHC II
Th1
Exotoxins
APC
atopic dermatitis CCL2
Exotoxins
TLR2 ligand
IL-31RA
Th2
IL-4
IL-13
IL-31
Keratinocytes
Sadaf Kasraie
Division of Immunodermatology and Allergy Research