Milan Plenary Epithelial response in atopy.ppt

The epithelial inflammatory response in an atopic environment

1. The epithelial inflammatory
response to viral infection in an
atopic environment
Nikos Papadopoulos
Allergy Dpt, 2
nd
Pediatric Clinic,
University of Athens

2. A conflict of interest is any situation in which a speaker or immediate family members have interests, and those may cause a conflict
with the current presentation. Conflicts of interest do not preclude the delivery of the talk, but should be explicitly declared. These
may include financial interests (eg. owning stocks of a related company, having received honoraria, consultancy fees), research
interests (research support by grants or otherwise), organisational interests and gifts.
Disclosure
In relation to this presentation, I declare the following, real or
perceived conflicts of interest:
Advisories: ABBOTT, Novartis, Menarini
Lectures: MSD, Uriach, GSK, Allergopharma, Stallergens, Menarini
MEDA
Grants: Nestle, MSD

3. Outline
•Virus-induced asthma – Rhinovirus is the most
relevant pathogen
•A central role for the airway epithelium
–Either because of inherent defects
–Or because of atopic conditioning
•Reciprocal interactions between virus-induced
epithelial and immune cell effects
•Attempts to disentangle

4. Viral infections are the major
trigger for acute exacerbations
•Human Rhinoviruses (RVs) are the most
relevant pathogens
5%
95%
C
h
i
l
d
r
e
n
Ruuskanen et al. (2006)
0%
20%
40%
60%
80%
100%
Children Adults
Other
RSV
Parainfluenza
Influenza
Corona
Rhinovirus
Johnston et al. BMJ (1995) 310:1225
Nicholson et al. BMJ (1993) 307:982
Papadopoulos et al. Allergy 2011;66:458-468

5. Lung function & inflammation in
preschool virus-induced asthma

0
0.5
1
1.5
2
R

5
H
z

b
e
f
o
r
e

b
r
o
n
c
h
o
d
i
l
a
t
o
r

(
k
P
a

/

l
t

/

s
e
c
)

p=0.0025
p=0.500
p=0.0008
Baseline Day 0 Day 10 Day 30


0

10

20

30

40
F
E
N
O

(
p
p
b
)

Baseline Day 0 Day 10 Day 30
p=0.0001

p=0.078
p=0.0009
R5 Hz
eNO
Konstantinou, Papadopoulos et al. JACI 2013 Jan;131(1):87-93

6. RVs may also initiate asthma of
make it persist
Infants with a symptomatic RV
infection have considerably increase
risk to continue wheezing after 3 years
Lemanske et al. J Allergy Clin Immunol 2005;116:571
Medium
RV
RV-induced VEGF and other
remodeling factors
Psaras, Papadopoulops et al. J Allergy Clin Immunol 2006;117:291

7. Understanding pathophysiology
•RV infects the bronchial epithelium inducing
inflammation
RV in the bronchial epithelium
Grunberg et al. Clin Exp Allergy (2000) 30:1015
Upregulation of epithelial ICAM-1
after experim
ental infection in hum
ans
Papadopoulos et al. J Inf Dis (2000) 181:1875

8. What makes asthmatics develop
symptoms following these
‘physiological’ responses ?

9. •A possible inherent epithelial defect, as a
‘core’ pathophysiology in asthma
•A possible deviation in the epithelial response
induced by/associated with atopy

10. Supported by an FP7 European
Commission Grant
Athens NG Papadopoulos
London SL Johnston
Vienna R Valenta
SIAF M Akdis
BRF AthensV Andreakos, D Thanos
Lodz M Kowalski
Ghent C Bachert
Marburg H Renz
Turku T Jartti, T Vuorinen
ErlangenS Finotto
www.predicta.eu

11. Airway remodeling
•A failure in the epithelial
injury – repair cycle
–Increased susceptibility to
injury
–Impaired wound healing

12. RV infection delays epithelial repair
 




Baseline
24h-Control
24h-RV infected
Bossios et al. Resp Res 2005;6:114
0
200
400
600
800
1000
1200
0 24 48 72
Hours
S
q

p
i
x
e
l
s
Control
RV

13. Response to RV is defective in epithelial cells
from atopic asthmatics
•Lower apoptosis,
higher cytotoxicity
and RV-titre in cells
from asthmatics
Wark et al. JEM (2005) 201:937-47

14. An IFN-–related effect
Wark et al. JEM (2005) 201:937-47

15. Type-3 interferons
•Deficient induction in atopic asthmatics
Contoli et al Nature Med (2006)

16. Cytokine environment affects RV
epithelial infection
Subauste et al, John Hopkins

17. •Exposure of PBMC from normal or atopic
asthmatic individuals to RV
•Collection & characterization of supernatant
•Infection of epithelial cells with RV, in the presence
of the above supernatants
•Assessment of mediators, cytotoxicity, virus
replication
; 2008

18. Effect of an atopic environment on virus
replication
RV TITER
0
1
2
3
0 6 48HOURS
L
O
G
ATOPIC
NORMAL
Xatzipsalti et al. Clin Exp Allergy. 2008;38:466-72.

19. 3
5
7
9
1
1
l
o
g

(
p
g
/
m
l
)
IL-6
Effect of an atopic environment on epithelial
inflammation – IL6
((--)) RVRV Atopic Atopic Normal Normal
p<0.001
Xatzipsalti et al. Clin Exp Allergy. 2008;38:466-72.

20. Immune protection of epithelial cytotoxicity is
hampered in atopy
Control RV + atopic + normal
p=0.001
p<0.001
Xatzipsalti, Papadopoulos et al. Clin Exp Allergy 2008

21. RV-induced angiogenic factor VEGF is
increased in an atopic environment
Psaras et al J Allergy Clin Immunol 2006;117:291

22. RV-induced FGF-2 and TGF-β are
upregulated in an atopic environment
Normal AtopicAsthmatic
0
25
50
75
100
125
b
F
G
F

(
p
g
/
m
l
)
*
Clinical Translational Allergy 2012; 2;14

23. Do RV-induced epithelial responses influence
T-cell differentiation ?
Makrynioti, Akdis, Johnston, Papadopoulos et al 2013 - in preparation

24. RV-infected epithelial supernatant conditioning
of Th0 cells
Intracellular Levels, n=5

25. Attempts to disentangle…

26. Papi et al J Allergy Clin Immunol 2012;130:1307-14
•47 children (5 ± .5 yrs) undergoing
bronchoscopy
•Asthmatic, atopic or both
•Bronchial biopsy and epithelial cells
from brushings
•RV infection and evaluation of the
innate IFN responses, inflammation
JACI 2012;130:1307-14

27. Airway pathology – asthma and atopy
Baraldo et al. JACI 2012;130:1307-14

28. Variability
•Children vs adults
•Disease severity
•Steroid treated vs non-treated
•Upper vs lower airway epithelium
–Including temperature differences (33° vs 37°)

29. Temperature-dependent differential
susceptibility to RV of nasal, tracheal and
bronchial epithelium
Othumpangat et al., 2012 Am J Physiol Lung Cell Mol Physiol 302(10):L1057-66
Nasal, tracheal, and bronchial epithelial cell
lines
Infected with RV at 33°C or 37°C
HRV-16 copy number measured by Q-PCR

30. RV replicates more in bronchial than
nasal epithelial cells
•Primary nasal & bronchial cells
from atopic asthmatics and
normal individuals – ALI culture
Nasal
control
Nasal
asthmatics
Bronchial
control
Bronchial
asthmatics
Lopez-Souza et al., 2010
*
Papadopoulos et al unpublished

31. PreDicta WP3
•Can the IFN deficiency attributed to asthma or atopy?
•Are IFN defects also present in the nose?
•Exposure of primary nasal epithelial cells to RV
•Assessment of viral load, IFN-β expression
•Groups:
–Controls (no atopy, rhinitis or asthma)
–Allergic Rhinitis (no asthma)
–Allergic Asthma (with rhinitis)
–Non-allergic asthma (with rhinitis)

32. IFN-β expression is deficient in the
nasal epithelium of asthmatics
*
*
But not in allergic rhinitis !
Papadopoulos et al. 2013 unpublished

33. Also dependent on disease severity
Papadopoulos et al. 2013 unpublished

34. Viral proliferation correlates to IFN
responses
lo
g
1
0
Δ
Δ
C
t
Papadopoulos et al. 2013 unpublished

35. Viral loads correlate to IFN levels
Papadopoulos et al. 2013 unpublished

36. Cytotoxicity at 48 hours
%
Papadopoulos et al. 2013 unpublished

37. Conclusions
•Epithelial RV infection and the resulting
inflammation are central pathophysiological
events in asthma
•The antiviral epithelial responses are defective
in (atopic) asthma, with innate interferons
being key mediators
•Epithelial and immune responses regulate one
another

38. Conclusions
•An atopic environment results in epithelial
antiviral defects and skew towards
remodeling
•The (asthmatic) epithelium can shift T-cells
responses towards a Th2 phenotype
•Interferon dysregulation is also present in
allergic rhinitis, but possibly not as simple as a
‘deficiency’

39. Acknowledgements
•Akis Megremis
•Stella Taka
•Heidi Makrynioti
•Chrysanthi Skevaki

40. NoOrganization name P.I. Country
1University of Athens N.G. Papadopoulos Greece
2Imperial S.L. Johnston UK
3Mediszinische Universitaet WienR. Valenta Austria
4SIAF M . & C. Akdis Switzerland
5BRF Athens E. Andreakos, D. ThanosGreece
6Uniwersytet Medyczny w LodziM. Kowalski Poland
7Universiteit Gent C. Bachert Belgium
8Philipps Universitaet MarburgH. Renz Germany
9Turun Yliopisto T. Jartti Finland
10Universitaetsklinikum ErlangenS. Finotto Germany
11BIOMAY AG F. Stoltz Austria
12INSERM TRANSFERT D. Fasquel France
www.predicta.eu
Acknowledgements