ICI 2016_Abstract Book

13 / 827

13
International Congress of Immunology 2016 Abstract Book
into conditional MC-deficient Cpa3-Cre; Mcl-1fl/fl mice. In this in
vivo setting, loss of MC-Nedd4-2 or -Ndfip1 resulted in a striking
sustained inflammatory influx at the reaction site. Collectively,
our findings reveal a previously unknown negative regulatory
mechanism whereby Nedd4-2 and Ndfip1 are essential to
restrain MC function.
1908
Humanized allergic NOD-SCID IL-2Rγc-/- mice as in vivo
model for novel allergy vaccines
Vizzardelli, C.1 , Nagl, B.1, Neunkirchner, A.2, Zimmann, F.1,
Kitzmüller, C.1, Jahn-Schmid, B.1, Pickl, W.F. 2, Bohle, B.1
1Medical University Vienna, Department of Pathophysiology and
Allergy Research, Vienna, Austria,2Medical University Vienna,
Institute of Immunology, Vienna, Austria
Birchpollen allergy affects around100millionpeopleworldwide.
The only treatment curing IgE-mediated allergy with long-term
benefit is allergen-specific immunotherapy (AIT). Unfortunately,
AIT is not successful in all treated patients. Therefore, one major
focus of allergy research is the development of AIT vaccines
with improved efficacy. To study such vaccines in an in vivo
model imitating the human allergic response we humanized
NOD-SCID IL-2Rγc-/- (NSG) mice with cells from birch pollen-
allergic patients. Briefly, PBMC were injected intraperitonally
(i.p.) together with birch pollen extract (BPE). After seven
days, mice were boosted once i.p. with BPE. In addition, mice
were humanized with BPE-specific T cell lines (TCL) expanded
from PBMC of birch pollen-allergic patients. NSG-mice were
challenged thrice intranasally (i.n.) with BPE or PBS. Thereafter,
airway hyperresponsiveness (AHR) and lung inflammation was
monitored. Human CD45+ cells were assessed in lungs and
spleens by flowcytometry.
MicehumanizedwithPBMCandBPE-specificTCL and challenged
with BPE showed significantly higher AHR and higher numbers
of eosinophils, neutrophils, and basophils, in bronchoalveolar-
lavage-fluids (BALF) than mice challenged with PBS. Human
CD45+ cells were detected in lungs and spleens in both
humanization conditions. As expected, higher percentages of
CD4+ T cells were found in mice humanized with BPE-specific
TCL. NSG-mice humanized with cells from birch pollen-allergic
donors developed allergic lung inflammation to birch pollen.
We will now use this in vivo model for testing of novel vaccines
for birch pollen allergy developed in our laboratory.
Supported by Austrian Science Fund, projects SFB 4609, 4610
and W1248
3791
Role of interleukin-36γ in regulating lung inflammation
Tay, H., Yang, M., Hsu, A., Nguyen, T.-H., Plank, M., Maltby, S.,
Bartlett, N., Hansbro, P., Foster, P.
The University of Newcastle, Immunology & Microbiology,
Newcastle, Australia
Respiratory infections are thought to be one of the major risk
factors that trigger the steroid resistant asthma exacerbations.
In these patients innate inflammatory mediators and cells are
linked to pathogenesis and disease severity. Our investigations
have identified novel roles of interleukin (IL)-36 family
cytokines in regulating airway inflammation and alterations
in lung function during infection and pathogen-induced
exacerbation of asthma. We found that the expression of IL-36γ
was significantly increased by pathogen-associated molecular
patterns derived from bacteria and viruses in macrophages.
Administration of recombinant IL-36γ in vivo leads to increase
inflammatory genes expression (gene profiling) and recruitment
of innate immune cells that are linked to asthma pathogenesis.
We next sought to determine the importance of IL-36γ in asthma
using a well-characterised murine model of allergic airway
disease. Mice were sensitised and challenged with ovalbumin
(OVA) to induce hallmark features of asthma. During allergen
challenge, recombinant IL-36γ was administered intranasally
and pulmonary function was assessed. Mice treated with IL-36γ
became insensitive to corticosteroid therapy. In conclusion,
we show that infections that trigger asthma exacerbations are
linked to IL-36productionand that deliveryof this cytokine to the
airways results in steroid-resistant airway hyperresponsiveness
and neutrophilic inflammation. Our results suggest that IL-36γ
could be a potential therapeutic target for neutrophilic and
severe asthmatics.
2813
Parvalbumin from Atlantic cod interacts with bronchial
epithelial cells and induces differential expression of
cytokines
Kalic, T.1, Ellinger, I.1, Gepp, B.1, Radauer, C. 1, Waltl, E.2, Niederberger,
V.2, Breiteneder, H.1
1 Medical University of Vienna, Department of Pathophysiology and
Allergy Research, Vienna, Austria, 2Medical University of Vienna,
Department of Otorhinolaryngology, Vienna, Austria
Background: Inhalation of aerosolized fish allergens and fish
matrix components is often associatedwith severe IgE-mediated
reactions in sensitized individuals. We explored interactions of
bronchial epithelial cells with the major codfish allergen Gad m
1 and the cod food matrix.
Methods: We used the human bronchial epithelial cell line
16HBE14o-. Polarized cells were treated with natural Gad m 1
with or without the codfish-derived food matrix. Fluorescently
labelled allergen was detected by confocal microscopy.
mRNA levels of IL-6 and CCL2 were determined by qRT-PCR.
Concentration of cytokines in the basolateral cell culture
medium was measured by the Luminex assay.
Results: Apical exposure of the cells to Gad m 1 resulted in
binding of the allergen to the lateral membrane domain (bellow
ZO-1 level). The allergen was not internalized by the cells nor
detected in the basolateral medium. IL-6 mRNA and protein
levels were increased by 30% after treatment with codfish
matrix but not with the allergen alone. The CCL2 concentration
was 50% higher in the basolateral medium of samples treated
with Gad m 1 compared to fish matrix-treated samples and the
negative control.
Conclusion: The major codfish allergen Gad m 1 interacts
with the plasma membrane of bronchial epithelial cells. This
interaction leads to changes in gene and protein expression