Papadopoulos, Nikolaos

Skevaki, Chrysanthi

Spyridaki, Irini

Taka, Styliani

Trochoutsou, Aikaterini

Clinical and Translational Allergy

Irini Spyridaki

Chrysanthi Skevaki

Aikaterini Trochoutsou

Styliani Taka

Nikolaos Papadopoulos

Springer - Publisher Connector

ORAL PRESENTATION Open Access
O04 - Effects of 5-lipoxygenase pathway
inhibition on rhinovirus-associated bronchial
epithelial inflammation
Irini Spyridaki*, Chrysanthi Skevaki, Aikaterini Trochoutsou, Styliani Taka, Nikolaos Papadopoulos
From 3rd Pediatric Allergy and Asthma Meeting (PAAM)
Athens, Greece. 17-19 October 2013
Background
Human bronchial epithelial cells produce a variety of
inflammatory mediators upon exposure to rhinovirus
(RV), a major precipitant of asthma exacerbations. We
hypothesized that anti-leukotriene (LT) treatment of
epithelial cells with or without exposure to supernatants
of RV-infected peripheral blood mononuclear cells
(PBMCs) may inhibit RV-induced up-regulation of
inflammatory cytokines.
Methods
PBMCs were isolated from a non-atopic, non-asthmatic
donor and exposed to 1MOI of RV-1B, and supernatants
were harvested at 48h post-infection. Subsequently,
BEAS-2B, a bronchial epithelial cell line, was infected
with RV, with or without conditioning with PBMC super-
natants. Treatment with anti-LT agents was performed
either on both PBMCs and BEAS-2B or at the bronchial
epithelial level only, with varying concentrations of mon-
telukast or MK-886. We evaluated the concentration of
inflammatory cytokines (IL-8, RANTES, IL-11, IL-6 and
IP-10) in culture supernatants at 48 hours after infection
of BEAS-2B cells with the use of commercially available
immunoassays.
Results
Treatment of PBMCs and BEAS-2B with montelukast at
concentrations of 10-4M to 10-6M, and with MK-886 at a
concentration of 10-4M significantly inhibited release of
IL-8 by RV-infected BEAS-2B. RANTES, IL-6 and IP-10
release was inhibited at all concentrations tested by both
drugs, while IL-11 release was inhibited only after
treatment with montelukast. Treatment of BEAS-2B cells
with montelukast or with MK-886 at a concentration
of 10-6M inhibited release of all cytokines measured,
irrespective of exposure to conditioned media of RV-
infected PBMCs, while 10-9M montelukast inhibited the
release of IL-8, IL-11 and IL-6, and 10-9M MK-886 sup-
pressed the release of IL-8 and RANTES.
Conclusion
Our results show that anti-LT treatment of RV-infected
bronchial epithelial cells, with or without exposure to con-
ditioned media of RV-infected PBMCs suppresses epithe-
lial RV-mediated inflammatory cytokine production. Our
observations may represent an indirect mode of action of
anti-leukotriene medication in virus-induced asthma.
Published: 28 February 2014
doi:10.1186/2045-7022-4-S1-O4
Cite this article as: Spyridaki et al.: O04 - Effects of 5-lipoxygenase
pathway inhibition on rhinovirus-associated bronchial epithelial
inflammation. Clinical and Translational Allergy 2014 4(Suppl 1):O4.
Allergy Research Laboratories, Second Department of Pediatrics, University of
Athens, Athens, Greece
Spyridaki et al. Clinical and Translational Allergy 2014, 4(Suppl 1):O4
http://www.ctajournal.com/content/4/S1/O4
© 2014 Spyridaki et al; licensee BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative
Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and
reproduction in any medium, provided the original work is properly cited. The Creative Commons Public Domain Dedication waiver
(http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.


O04 - Effects of 5-lipoxygenase pathway inhibition on rhinovirus-associated bronchial epithelial inflammation

Digital Repository of Hellenic Managing Authority of the Operational Programme &quot;Education and Lifelong Learning&quot; (EDULLL)

Effects of 5-lipoxygenase pathway inhibition on rhinovirus-associated bronchial epithelial inflammationIrini Spyridaki, Chrysanthi Skevaki, Aikaterini Trochoutsou, Styliani Taka (presenting), Nikolaos PapadopoulosAllergy Research Laboratories, Second Department of PediatricsUniversity of Athens This research has been co-financed by the European Union (European Social Fund – ESF)and Greek national funds through the Operational Program "Education and LifelongLearning" of the National Strategic Reference Framework (NSRF) - Research FundingProgram: Heracleitus II. Investing in knowledge society through the European Social Fund.This was an Investigator initiated study, also funded by Merck & Co., IncRVs and asthma exacerbationsRuuskanen et al. (2006)Wark et al. ERJ (2002) AdultsChildrenNG Papadopoulos and CL Skevaki. Asthma: An atlas of investigation and management, 1st ed, 2006RVs induce inflammation and remodelingLeukotriene pathway and inflammationHypothesisAnti-leukotriene treatment of epithelial cells with or without exposure to supernatants of RV-infected PBMCs may inhibit RV-induced up-regulation of inflammatory cytokines.Methods• PBMCs isolation: healthy donor – non atopic-non astmatic• BEAS-2B cell culture• In-vitro infection of BEAS-2B cells and/or PBMCs with A1B• In-vitro treatment of BEAS-2B cells and/or PBMCs  with Montelukast (cysLT receptor antagonist)or MK-886 (FLAP inhibitor)• Incubation at 33°C, 5% CO2 for 48 hours• Evaluation of the concentration of inflammatory cytokines: IL-8, RANTES, IL-11, IL-6, IP-10Results(I) IL-8050010001500200025003000IL-8 (pg/ml)BEAS-2b, IL-8 release (drug treated PBMCs)************ *** ***050010001500200025003000350040004500B2b-/PBMCs- B2b+/D/PBMCs+ B2b+/M6/PBMCs+ B2b+/M9/PBMCs+ B2b+/K6/PBMCs+ B2b+/K9/PBMCs+IL-8 (pg/ml)BEAS-2b, IL-8 release (PBMCs, drug treated)*** **********Results(II) RANTES05000100001500020000250003000035000RANTES (pg/ml)BEAS-2b, RANTES release (drug treated PBMCs)***** **********050001000015000200002500030000B2b-/PBMCs- B2b+/D/PBMCs+ B2b+/M6/PBMCs+ B2b+/M9/PBMCs+ B2b+/K6/PBMCs+ B2b+/K9/PBMCs+RANTES (pg/ml)BEAS-2b, RANTES release (PBMCs, drug treated)****** *****Results(III) IL-6010002000300040005000600070008000IL-6 (pg/ml)BEAS-2B, IL-6 release (drug treated PBMCs)*** *** *** ********** ***020004000600080001000012000IL-6 (pg/ml)BEAS-2B, IL-6 release (PBMC, drug treated)*** ** **Results(V) IP-10050010001500200025003000IP-10 (pg/ml)BEAS-2B, IP-10 release (drug treated PBMCs)**** ** ********05001000150020002500IP-10 (pg/ml)BEAS-2B, IP-10 release (PBMCs, drug treated)*********Results(IV) IL-110100200300400500600700IL-11 (pg/ml)BEAS-2b, IL-11 release (drug treated PBMCs)****** *020406080100120140B2b-/PBMCs- B2b+/D/PBMCs+ B2b+/M6/PBMCs+ B2b+/M9/PBMCs+ B2b+/K6/PBMCs+ B2b+/K9/PBMCs+IL-11 (pg/ml)BEAS-2b, IL-11 release (PBMCs, drug treated)*** ** *** *ConclusionAnti-leukotriene treatment of RV-infected bronchialepithelial cells suppresses epithelial RV-mediatedpro-inflammatory (IL-6, IL-8, RANTES, IP-10) andanti-inflammatory (IL-11) cytokine response.These observations may represent an indirectmode of action of anti-leukotriene medication invirus-induced asthma.