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Abstracts from the 3rd International Severe Asthma Forum (ISAF)

Abstracts from the 3rd International Severe Asthma Forum (ISAF) Clin Transl Allergy 2017, 7(Suppl 2):14 Clinical and DOI 10.1186/s13601-017-0149-8 Translational Allergy Open Access MEE TING ABSTR AC TS Abstracts from the 3rd International Severe Asthma Forum (ISAF) Manchester, United Kingdom. 17 November 2016–19 November 2016 Published: 24 May 2017 ORAL ABSTRACT SESSION 1—Asthma: from mechanisms (IL-1RL1-a alone: AUC = 0.65 [95 CI 0.52–0.79, P = 0.04], API + IL-1RL1- to management a: AUC = 0.70 [95 CI 0.56–0.84, P = 0.01]). Interestingly, IL-1RL1-a levels had a negative direction of effect. Conclusion: Our study shows that serum IL-1RL1–a levels measured in O01 wheezing children at age 2–3 years do not predict doctors’ diagnosed Serum IL‑1RL1‑A levels predict an eosinophilic subtype of asthma asthma as general phenotype at age 6 years, but negatively predict an in preschool wheezing children 1 2 1 3 eosinophilic subphenotype of asthma. M. E. Ketelaar , K. Van De Kant , F. N. Dijk , E. M. M. Klaassen , N. 4 4 2 1 This suggests that IL-1RL1 might play a protective role in the devel- Grotenboer , M. C. Nawijn , E. Dompeling , G. H. Koppelman 1 opment of eosinophilia in children who experience asthma at school University Medical Center Groningen, Beatrix Children’s Hospital, Gron- age and implies that IL-1RL1 targeted therapy could rather be further ingen Research Institute for Asthma and COPD (GRIAC), Groningen, The 2 explored in the subphenotype of asthmatic children with predomi- Netherlands; Department of Pediatric Pulmonology, School for Public nant eosinophilic inflammation. Health and Primary Care (CAPHRI), Maastricht University Medical Center 3 Keywords: Childhood Asthma, Eosinophilic Asthma, Prediction, (MUMC+), Maastricht, The Netherlands; Department of General Practice, IL-1RL1, Serum School for Public Health and Primary Care (CAPHRI), Maastricht University Medical Center (MUMC+), Maastricht, The Netherlands; University Medi- cal Center Groningen, Department of Pathology and Medical Biology, Laboratory of Experimental Pulmonology and Inflammation Research (EXPIRE), Groningen Research Institute for Asthma and COPD (GRIAC), Groningen, The Netherlands Correspondence: Maria Elizabeth Ketelaar - m.e.ketelaar@student.rug.nl Clinical and Translational Allergy 2017, 7(Suppl 2):O01 Introduction: Respiratory symptoms are common in preschool chil- dren. However, which of these wheezers will develop asthma at school age, and what phenotype they will develop remains difficult to pre - dict. Current models such as the asthma prediction index (API) are based on clinical parameters and have only modest predictive accu- racy. Expression levels of well replicated asthma genes could poten- tially form novel biomarkers for asthma prediction. IL1RL1 is an asthma susceptibility gene, and has also been linked to eosinophilia. Therefore, we hypothesized that expression levels of IL1RL1 in the form of soluble IL-1RL1-a measured in serum from wheezing preschool chil- dren contribute to the prediction of asthma at school age. Moreover, since IL1RL1 was previously associated with blood eosinophilia, our second aim was to determine whether serum IL-1RL1-a levels predict eosinophilic asthma. Method: We used logistic predictive modeling in a prospective Dutch Figure 1 Prediction of eosinophilic childhood using IL-1 RL1-a serum birth cohort (n  =  202 wheezers), and calculated the area under the levels and the API curve (AUC) of the sensitivity/1-specificity curves of potential models. Results: Neither IL-1RL1-a serum levels at age 2–3  years alone nor its combination with the API had predictive value for doctors’ diag- O03 nosed asthma at age 6y (IL-1RL1-a alone: AUC = 0.50 [95 CI 0.41–0.59, Diagnosing asthma in symptomatic children using lung function: P = 0.98], API + IL-1RL1-a: AUC = 0.57 [95 CI 0.49–0.66, P = 0.12]). evidence from a birth cohort study However, IL-1RL1-a serum levels at age 2–3  years correlated with the 1 1 1 1 Clare Murray , Philip Foden , Lesley Lowe , Hannah Durrington , severity of airway eosinophilia (determined by levels of exhaled frac- 2 1 Adnan Custovic , Angela Simpson tion of NO, [FeNO]) in children who had developed asthma at age 6y 1 2 University of Manchester, Manchester, United Kingdom; Imperial (Pearson’s R  =  −0.24, P  =  0.046, N  =  59). Logistic predictive mod- College, London, United Kingdom eling of eosinophilic asthma at age 6y (asthma with FeNO ≥  20  ppb) Correspondence: Angela Simpson - angela.simpson@manchester.ac.uk showed that IL-1RL1-a serum levels itself and in combination with Clinical and Translational Allergy 2017, 7(Suppl 2):O03 the API could predict this eosinophilic subphenotype of asthma © The Author(s) 2017. This article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. 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. Clin Transl Allergy 2017, 7(Suppl 2):14 Page 2 of 17 Introduction: In the UK, new national draft guidance for the diagno- Hospital of South Manchester, NHS Foundation Trust, Manchester, United sis of childhood asthma proposes algorithms based on four tests of Kingdom; Respiratory Research Unit, University of Nottingham, Notting- lung function, each used as a dichotomous variable (FE V /FVC ratio ham, United Kingdom; Respiratory Therapeutic Unit, GSK, Stockley Park, less than the lower limit of normal [LLN], bronchodilator reversibility London, United Kingdom; National Heart and Lung Institute, Imperial [BDR] ≥ 12%, FeNO ≥ 35 ppb and PEFR variability). However, accuracy College, London, United Kingdom; NIHR Southampton RespiratoryBio- of these tests in diagnosing asthma in children is unknown, as the medical Research Unit, Clinical and Experimental Sciences and Human evidence is largely derived from studies of adults. Within the setting Development and Health, Southampton, United Kingdom; Data Science of a population-based birth cohort (Manchester Asthma and Allergy Institute, South Kensington Campus, Imperial College London, London, Study—MAAS), we investigated the value of FE V /FVC, BDR and FeNO United Kingdom; Acclarogen Ltd, St John’s Innovation Centre, Cam- 8 9 in diagnosing asthma in children. bridge, United Kingdom; AstraZeneca R&D, Mölndal, Sweden; BioSci Method: Using validated questionnaires we assessed study par- Consulting, Maasmechelen, Belgium; Dept of Respiratory Medicine, ticipants at age 16  years. Current asthma was defined as all three Academic Medical Centre, University of Amsterdam, Amsterdam, The of: (1) doctor-diagnosed asthma ever, (2) wheezing in the previous Netherlands 12 months and (3) current use of asthma treatment. We assigned chil- Correspondence: Andrew J. Simpson - Andrew.Simpson-2@Manchester. dren negative to all three features as non-asthmatic controls. Using ac.uk ATS/ERS guidelines, we measured spirometry and FeNO (NIOX chemi- Clinical and Translational Allergy 2017, 7(Suppl 2):O04 luminescence analyser; Sweden). BDR was considered positive if FEV increased by > 12% following administration of 400 mg of salbutamol. Introduction: Individuals with severe asthma may remain uncon- PEFR variability was not measured. To test the diagnostic algorithms trolled and exacerbation-prone despite intensive guideline-directed simulating the clinic situation, we selected only children reporting treatment, and management options are limited. The concept of treat- recent symptoms of wheeze, cough or breathlessness who were not able traits, based on the identification of treatable disease-associated on regular inhaled corticosteroids (ICS). characteristics, may thus be a particularly useful framework in this Results: Of the 630 MAAS children with full data available, 163 context. In the Unbiased Biomarkers for the Prediction of Respiratory reported recent symptoms, but were not using regular ICS; 34 of Disease Outcomes (U-BIOPRED) project we have recruited individuals these met our definition of current asthma, with 55 as non-asth- with severe asthma (SA) under specialist care, and controls with mild matic controls. In the multivariable logistic regression analysis, to moderate asthma (MMA). increasing FeNO was associated with an increased risk of asthma Aim: To identify and quantify treatable traits within the U-BIOPRED (OR 1.02, 95% CI 1.01–1.04, p  =  0.006), with a trend for FE V /FVC adult asthma cohorts. ratio (OR 0.95, 95% CI 0.87–1.02, p  =  0.17), and no association for Method: We defined criteria for treatable traits based on Agusti (Eur BDR (p  =  0.94). The proportion of those with each combination of Respir J, 2016) and identified prevalence rates within the U-BIOPRED positive tests is show as a Venn diagram (Figure  1). Of 58 children database. Chi Square tests were used to examine differences in fre - with three negative tests, 29.3% had current asthma, accounting quency between individuals with SA and MMA for 50% of those with asthma. Only 5.9% of those with asthma were Results: Data from 509 individuals with asthma were included in the positive to all three tests. analysis; 421 with SA and 88 with MMA. Twenty-nine treatable traits were Conclusion: Applying 3 tests of lung function to children with symp- identified, including 13 pulmonary, 13 extra-pulmonary and three behav - toms and a diagnosis of asthma failed to detect 50% of asthma cases. ioral traits. Pulmonary treatable traits such as airflow limitation (SA 50% Proposed algorithms for the diagnosis of asthma in symptomatic chil- vs. MMA 6%, P  <  0.001), reversibility (SA 58% vs. MMA 39%, P =  0.002), dren need to be tested prospectively. eosinophilia (SA 54% vs. MMA 43%, P = 0.067), exercise-induced asthma Keywords: Asthma, Diagnosis, FeNO, Lung Function, Children (SA 77% vs. MMA 54%, P  =  0.002), allergic rhinitis (SA 47% vs. MMA 44%, P = 0.641), cough (SA 63% vs. MMA 19%, P < 0.001) and bronchitis (SA 51% Vs. MMA 16%, P  =  0.000) were highly prevalent in the asthma cohorts, and typically more common in SA versus MMA. The most com- mon extra-pulmonary treatable traits were; atopy (SA 74% vs. MMA 92%, P < 0.001), obesity (SA 30% vs. MMA 38%, P = 0.178), reflux (SA 36% vs. MMA 11%, P  <  0.001), hypertension (SA 25% vs. MMA 9%, P  =  0.001) and obstructive sleep apnea (SA 26% vs. MMA 11%, P  =  0.003). Behav- ioral traits included low medication adherence (SA 39% vs. MMA 52%, P = 0.031) and smoking (SA 8%, smokers not eligible in MMA cohort). Par- ticipants with SA had mean (SD) 8 ± (2), and MMA 5 ± (2) treatable traits. Conclusion: We have applied a new approach for the characterisation of severe asthma, based on treatable traits. In general these traits were found more commonly in severe than non-severe asthma; the very high prevalence of many of these traits suggests that there are poten- tial targets for treatment even in such severe patients, and supports the need for specialist management. Keywords: Phenotypes, Treatable Traits Figure 2 Venn diagram showing number of children with symptoms (n163) who were positive for each combination of tests ORAL ABSTRACT SESSION 2—Molecular mechanisms O05 O04 Discovery, development and validation of blood‑based RNA Treatable traits in the European U‑BIOPRED adult severe asthma biomarker panels to predict the late‑phase asthmatic response 1 1 2 1 1 cohort S. J. Tebbutt , A. Singh , C. P. Shannon , Y. W. Kim , C. X. Yang , G. M. 1 2 3 4 3 1 4 3 Andrew J. Simpson , Dominick E. Sha w , Ana R. Sousa , Louise J. Fleming , Gauvreau , J. M. Fitzgerald , L. P. Boulet , P. M. O’Byrne 5 6 7 8 1 2 Graham Roberts , Ioannis Pandis , Aruna T. Bansal , Julie C orfield , Scott University of British Columbia, Vancouver, Canada; PROOF Centre 9 5 4 10 3 Wagers , Ratko Djukanovic , Kian Fan Chung , Peter J. Sterk , Jorgen of Excellence, Vancouver, Canada; McMaster University, Hamilton, 1 1 4 Vestbo , Stephen J. Fowler Canada; Université Laval, Québec City, Canada Division of Infection, Immunity and Respiratory Medicine, School Correspondence: Scott J. Tebbutt - scott.tebbutt@hli.ubc.ca of Biological Sciences, The University of Manchester and University Clinical and Translational Allergy 2017, 7(Suppl 2):O05 Clin Transl Allergy 2017, 7(Suppl 2):14 Page 3 of 17 Introduction: We have previously demonstrated significant molecu- Measurements of dynamic resistance were performed using a Flex- lar changes in the blood between mild allergic asthmatic individuals ivent system (Scireq, Montreal, Canada). After induction of anaesthesia who develop isolated early responses (early responders, ERs) com- with an intrperitoneal injection of ‘Hypnorm’ (0.315 mg fentanyl; 10 mg pared to those who also develop late-phase responses (dual respond- fluanisone) and midazolam (5  mg) in water, at a dose of 0.1  ml/10  g, ers, DRs) after allergen inhalation challenge. Identifying individuals mice were tracheostomized and connected to the flexivent ventilator likely to develop dual responses may aid in the screening of subjects via an 18-guage needle. Mice were ventilated. Changes in resistance for clinical trials that test drugs for the attenuation of the late-phase were measured in response to increasing concentrations of nebulized asthmatic response, which shares hallmark features of chronic disease. methacholine from 0 to 300  mg/ml. All measurements were made in The objective of this study was to develop blood-based biomarker the morning. −/− panels that could identify asthmatic individuals with high probability Results: Rev-erb alpha mice exhibit significantly (p  <  0.01) greater of developing a late-phase response. AHR to increasing concentrations of methacholine compared to their Method: The discovery cohort consisted of 36 mild asthmatic subjects wildtype and heterozygote litter-mate controls (Figure 1). −/− (15 ERs and 21 DRs) and a validation cohort consisted of 45 mild asth- Conclusion: Rev-erb alpha mice retain circadian rhythmicity yet matic subjects (9 ERs and 36 DRs). Blood samples were collected prior demonstrate increased AHR to methacholine (as is seen in human to allergen challenge. Following RNA extraction, total RNA (globin-/ asthma). REV-ERB alpha may play an important role in the pathogen- ribo-depleted) was sequenced using an Illumina HiSeq 2000 as 100 esis of asthma. base paired-end reads. Both genome-guided datasets (UCSC genes, Next steps: UCSC gene-isoforms, and Ensembl) and de novo assembled transcripts using the Trinity software were constructed. Top-ranked biomarker −/− 1. Investigate a time of day difference in AHR in Rev -erb alpha candidates were transferred to the high-precision and clinically- mice approved NanoString nCounter platform. Final biomarker panels were −/− 2. Expose Rev-erb alpha mice to the house dust mite model of identified, and statistical algorithms were locked down prior to testing asthma in the validated cohort. 3. Employ REV-ERB alpha ligands to ‘modify’ the phenotype Results: Predictive biomarker panels had classification performance (based on the area under the receiver operating curve, AUC) rang- Keywords: Airway Hyperresponsiveness, Rev-Erb Alpha, NR1D1, Clock ing between 60 and 70% in the discovery cohort. 87 transcripts Gene identified on the RNA-Seq platform were transferred to NanoString Elements assay chemistry. The transcripts were split with respect to their dataset of origin and tested in the validation cohort. The UCSC gene-isoforms and Trinity biomarker panels had AUCs of 68% and 72%, respectively. The biomarker transcripts were enriched for bio- logical pathways such as NF-ĸB signaling (up-regulated in DRs) and apoptosis, decoy receptors and formyl peptide receptors (down-reg- ulated in DRs). Conclusion: RNA transcript biomarker panels in the blood were successful at predicting asthmatic subjects likely to develop dual responses upon allergen inhalation challenge. Pathways and networks represented by these biomarker panels may reveal new avenues for therapeutic targeting of the pathobiology involved in chronic asthma. Keywords: Predictive Biomarkers, RNA-Seq, NanoString Elements, Sta- tistical Algorithms O06 The clock Gene REV‑ERBalpha regulates airway Figure 3 Airway Hypersensitivity to Methacholine hyperresponsiveness: implications for asthma H. J. Durrington, N. Begley, A. Loudon, D. W. Ray University of Manchester, Manchester, United Kingdom Correspondence: Hannah J. Durrington - hannah.durrington@manches- O07 ter.ac.uk Asthma biomarkers: methylation and correlation with functional Clinical and Translational Allergy 2017, 7(Suppl 2):O06 parameters 1 1 1 2 Selene Baos , Lucía Cr emades , David Calzada , Carlos Lahoz , Blanca Introduction: Asthma is an inflammatory disease with a strong circa- Cárdaba 1 2 dian signature. Symptoms of asthma are worse overnight and in the IIS-Fundación Jiménez Díaz-UAM, Madrid, Spain; IIS-Fundación Jiménez early hours of the morning. Measurements of peak expiratory flow Díaz-UAM; Ciber de Enfermedades Respiratorias, CIBERES, Madrid, Spain rate and forced expiratory volume in 1  s, are lower in early morn- Correspondence: Selene Baos - selene.baos@fjd.es ing compared to afternoon. It is likely that circadian biology plays Clinical and Translational Allergy 2017, 7(Suppl 2):O07 a role in the pathogenesis of asthma. A better understanding of the circadian nature of asthma may lead to better treatments through Introduction: In a previous study (submitted to publish) we defined chronotherapy (taking medication at the most beneficial time of day). specific genes related with asthma and allergic diseases, by study - Clock genes control circadian rhythms within every cell in the body. ing the gene-expression of 94 genes in a population composed by 4 The clock gene, REV-ERB alpha (NR1D1), connects the ‘core’ clock and groups of subjects: healthy control, nonallergic asthmatic, asthmatic the immune system. We employed a REV-ERB alpha global knockout allergic and nonasthmatic allergic patients. The analysis of differen- −/− mouse (rev-erb alpha , these mice retain circadian rhythmicity, but tial gene-expression between control and patients with nonallergic show loss of temporal variation of innate immunity) to investigate the asthma revealed a set of statistically relevant genes mainly associated effect of REV-ERB alpha on airway hyper-responsiveness (AHR). AHR is with the disease’s severity: IL10, MSR1, PHLDA1, SERPINB2, CHI3L1, a key pathophysiological feature of asthma. IL8, and PI3. In this project we analyzed if methylation takes part in Method: All experimental procedures were carried out in accordance the regulation of the gene-expression of these potential asthma bio- −/− with the Animals (Scientific Procedures) Act,1986. Rev-erb alpha markers and their correlation with protein expression and functional mice were provided by Ueli Schibler (University of Geneva). parameters (%FVE1,  %FVC and  %PBD). Clin Transl Allergy 2017, 7(Suppl 2):14 Page 4 of 17 Method: DNA extracted from PBMCs of control and nonallergic O09 asthma subjects was treated with sodium bisulfite and amplified by Hyaluronan‑induced lipid mediators influence antiviral PCR with primers designed to amplify CpG islands near the promotor and antibacterial immunity in severe asthmatics 1 2 2 region of the most significant genes: MSR1 (5 CpG islands), SERPINB2 Milena Sokolowska , Li-Yuan Chen, Yueqin Liu , Asuncion 2 2 2 3 (5), PHLDA1 (35), CHI3L1 (8) and PI3 (5). The methylation analysis was Martinez-Anton , Carolea Logun , Sara Alsaaty , Rosemarie Cuento , 2 2 4 5 done with the Sequenom EpiTYPER approach. Protein quantification Rongman Cai , Junfeng Sun , Oswald Quehenberger , Aaron Armando , 5 3 2 was determined by ELISA or Western Blot. Statistical analyses were Edward Dennis , Stewart Levine , James Shelhamer performed with Graph-Pad program. Critical Care Medicine Department, Clinical Center, NIH; Swiss Institute Results: The methylation analysis showed statistically significant dif- of Allergy and Asthma Research, University of Zurich; CK-CARE, Davos, ferences between groups in 4 genes: MSR1, PHLDA1, CHI3L1 and PI3. Switzerland; Critical Care Medicine Department, Clinical Center, NIH, The results of the methylation correlated with the gene-expression, Bethesda, Maryland, United States; Laboratory of Asthma and Lung the protein levels and respiratory parameters, specifically in MSR1, Inflammation, Cardiovascular and Pulmonary Branch, National Heart, CHI3L1 and PI3. Lung and Blood Institute, NIH, Bethesda, Maryland, United States; Conclusion: A possible regulatory mechanism of molecular biomark- Department of Medicine, Department of Pharmacology, San Diego, ers of asthma has been defined, being methylation a possible key fac - California, United States; Department of Chemistry and Biochemistry, tor for the differential gene-expression of asthma patients. San Diego, California, United States Keywords: DNA Methylation, Biomarkers, Epigenetics Correspondence: Milena Sokolowska - milena.sokolowska@siaf.uzh.ch Clinical and Translational Allergy 2017, 7(Suppl 2):O09 O08 Critical role for Arginine metabolism in a combined Th2 and Th17 Introduction: Hyaluronan (HA) is the major glycosaminoglycan in the airway inflammation in the house dust mite model extracellular matrix involved in the pathogenesis of asthma and other Kewal Asosingh, Chris Lauruschkat, Kimberly Queisser, Nicholas Wanner, chronic inflammatory diseases. During inflammation in asthma there is Kelly Weiss, Weiling Xu, Serpil Erzurum an increased breakdown of HA, resulting in the local and systemic accu- Cleveland Clinic, Cleveland, Ohio, United States mulation of low molecular weight (LMW) HA. Eicosanoids, derived from Correspondence: Kewal Asosingh - asosink@ccf.org cytosolic phospholipase A group IVA (cPLA alpha) activation, are potent 2 2 Clinical and Translational Allergy 2017, 7(Suppl 2):O08 lipid mediators also attributed to acute and chronic inflammation. Method: We investigated the effect of LMW HA on the lipidomic pro - file and global gene expression and their functional interactions in Introduction: Inducible nitric oxide synthase (iNOS) and arginase-2 peripheral blood mononuclear cells of patients with mild-to-moderate (ARG2) share a common substrate arginine. We recently showed (n = 7) and severe asthma (n = 6) as compared to controls (n = 6). that both enzymes are highly expressed in asthmatic airway epi- Results: We found that LMW HA increased production of 68 unique thelial cells where ARG2 plays a critical role in the regulation of lipid species, among which PGE , PGB, PGD, 15-HETE, TxB , 11(12)- 2 2 2 2 mitochondrial bioenergetic driven Th2 response, as shown by the EET, 14(15)-EET, 13-HOTrE(y) and 16(17)-EpDPE were significantly development of severe eosinophilic airway inflammation in mice upregulated only in severe asthmatics. We also performed a genome- deficient for ARG-2 (ARG-2 KO). The role of iNOS and ARG2 in a com- wide expression analysis of LMW HA signaling, confirming its highly bined Th2 and Th17 response is unknown. Here we hypothesize immunostimulatory potential. However, in severe asthmatics the LMW that Th17 airway inflammation is also regulated by ARG2 driven HA-induced global gene expression profile showed a comprehen- metabolism. sive impairment in interferon signaling, cell apoptosis and cell move- Method: Commercially available Wildtype (WT) and iNOS knockout ment, leading to diminished antiviral and antibacterial responses. We (KO) were used. ARG2 KO was donated to us and ARG2iNOS double KO confirmed these findings at the protein level, finding that LMW HA- (dKO) strain was generated in house. Standard house dust mite extract induced production of IL-12 p40, CXCL10, CXCL11 and CCL8 in severe (HDME) mouse model of Th2/Th17 asthma endotype was used. Air- asthmatics was markedly reduced. Importantly, upon cPLA alpha inhi- way inflammation was quantified by analysis of inflammatory cells in bition, there was a significant decrease in lipid mediator production bronchoalveolar lavage fluid (BALF). Airway angiogenic remodeling accompanied by a significant increase in IL-12 p40 and CXCL9 protein was measured by quantification of lung microvessel density on tis- expressions in each phenotype. sue sections stained for the endothelial marker von Willebrand Factor. Conclusion: In summary, we demonstrated here that fragmented hya- Epithelial and immune cell cytokines were measured by ELISA or flow luronan increased production of several unique lipid species in severe cytometric multiplex assay. asthmatics. Moreover, through analysis of the whole genome pro- Results: HDME induced airway inflammation in all genotypes, but file, captured simultaneously with the lipidomic profile, we observed was the highest in ARG2 KO as showed by increased number inflam- decreases in antiviral gene and protein expression upon LMW HA treat- matory cells in the BALF. Both eosinophilic (Th2) and neutrophilic ment in severe asthmatics, which was partially reversed upon inhibition (Th17) inflammation peaked in ARG2 KO mice but were attenuated of lipid production. Therefore, our current findings provide new evidence in ARG2iNOS dKO animals. Airway inflammation was similar among on the connection between extracellular matrix, global lipid mediator HDME exposed WT, iNOS KO and ARG2iNOS dKO mice. Angiogenic production and decreased antiviral responses in severe asthma. airway remodeling was also highest in ARG2 KO mice and comparable Keywords: Severe Asthma, Eicosanoid, Virus, Lipidomics, among WT, iNOS KO and ARG2iNOS dKO genotypes. Airway epithelial Transcriptomics or immune cell-derived Th2 cytokines IL-5 and eotaxin-2 and, Th17 cytokine IL-17 were maximal in lungs of HDME exposed ARG2 KO mice, but decreased in ARG2iNOS dKO. POSTER DISCUSSION SESSION 1—Epidemiology Conclusion: The data show that ARG2 deficiency induces a severe combined Th2 and Th17 airway inflammation. Deletion of iNOS in P01 addition to ARG2 inhibited disease severity, but iNOS deletion alone, Fungal communities in house dust associated with the had no effect, indicating that ARG2 is the most critical of the two argi- development of Atopic Dermatitis in infant nine consuming enzymes in the regulation of inflammation and angi- Kilyong Choi ogenesis in asthma. Overall, the results expand our previous findings Seoul Medical Center, Seoul, South Korea by demonstrating that in addition to Th2, Th17 endotype of airway Correspondence: Kilyong Choi - bestchoi9494@gmail.com inflammation is also critically regulated by arginine metabolism Clinical and Translational Allergy 2017, 7(Suppl 2):P01 Keywords: Arginine, Metabolism, Th2, Th17, Mitochondria Clin Transl Allergy 2017, 7(Suppl 2):14 Page 5 of 17 Introduction: In our epidemiologic study, the effect of mold exposure Table 2 List of fungal genera and their relative abundance was associated with the exacerbation or the development of atopic der- (%) in house dust samples from two groups matitis (AD). Maternal exposure to mold during the pregnant periods or exposure to mold in early life particularly have an impact on the increased Taxon Status risk of AD development. However, its specific mechanism still remains Phylum Genus Health AD (n = 20) p Value unclear. In addition, dust is a repository and concentrator of many chemi- (n = 20) cal and biological agents including fungi. The aims of this study were to identify the fungal microbiota (mycobiota) in house dust, and to investi- Ascomycota Alternaria 7.9104 12.8797 0.079 gate whether the mycobiota is associated with the development of AD. Ascomycota Aspergillus 4.1237 5.1118 0.705 Ascomycota Aureobasidium 5.8441 5.0303 0.85 Table  1 Library coverage estimations and  sequence diver‑ Ascomycota Candida 0.7504 1.0536 0.725 sity of fungal ITS genes pyrosequencing Ascomycota Cladosporium 7.8394 7.6885 0.725 ID Valid OTUs Chao1 Shannon Simpson Goods Ascomycota Cyphellophora 0.5648 0.6081 0.022 reads Lib. Ascomycota Exophiala 0.3533 0.8299 0.957 Coverage Ascomycota Fusarium 0.6404 1.3015 0.685 S13-033-C 9475 474 769.5882 3.619653 0.114784 0.978786 Ascomycota Leptosphaer- 1.4300 0.8881 0.291 S12-073-C 8588 456 567.125 4.541252 0.02198 0.985212 ulina S12-067-C 10169 611 731.3051 4.629529 0.024318 0.983381 Ascomycota Meyerozyma 0.0367 1.7910 0.989 A13-033-C 9719 377 455.9153 3.690041 0.090025 0.99002 Ascomycota Nigrospora 0.7219 0.5520 0.344 A12-080-C-22 7111 315 404.7581 3.541059 0.09324 0.985094 Ascomycota Penicillium 2.7624 1.3681 0.978 A12-072-C 7899 439 757.3333 3.991715 0.051364 0.97582 Ascomycota Peyronellaea 1.6187 2.7516 0.818 A12-054-C 9467 343 574.6667 3.859098 0.05143 0.985212 Ascomycota Phoma 7.1393 6.3727 1 Y10-533-C 10273 520 775.6081 4.314013 0.038003 0.981018 Ascomycota Pleospora 0.4702 0.5722 0.487 C12-044-C 8788 382 518.1224 4.288241 0.030933 0.9868 Ascomycota Saccharomyces 2.9811 1.0517 0.176 Y12-016-C 11603 576 952.9615 4.045346 0.052539 0.979057 Basidiomycota Cryptococcus 1.8277 2.1603 0.725 A13-047-C 9386 517 740.9474 4.419917 0.037477 0.98029 Basidiomycota Malassezia 10.8815 4.4114 0.005 C13-065-C 11197 469 599.013 4.253351 0.033662 0.987318 A10-581-C 11167 530 693.4 4.24386 0.047244 0.984597 Basidiomycota Malassezi- 0.8520 1.2991 0.626 aceae_uc A10-609-C 12620 725 1126.857 4.347734 0.035431 0.976941 Basidiomycota Pleurotus 3.2915 0.0260 0.005 A12-037-C 11089 828 1181.979 4.64697 0.035575 0.970962 Basidiomycota Rhodotorula 1.5704 0.7741 0.725 A13-069-C 12210 764 1144.971 4.907156 0.02687 0.976904 Basidiomycota Trichosporon 4.1956 0.3016 0.003 C13-067-C 10215 570 733.8298 4.774492 0.021033 0.98277 S12-072-C 6413 317 375.2632 3.910256 0.058516 0.987213 Y12-044-C 12008 638 949.3542 4.230534 0.070634 0.979597 A12-071-C 4753 544 746.4118 5.021128 0.018621 0.960867 Method: To identify the mycobiota in house dust, DNA pyrosequenc- Y12-051-C 7282 329 416.3061 4.209385 0.030832 0.987229 ing was performed targeting the internal transcribed spacer (ITS) A10-575-C 9011 608 787.1 4.68377 0.025285 0.977916 region. Here, we investigated the composition of fungal communities in the dust samples from houses of 20 healthy and 20 AD infants diag- A12-074-C 10195 410 549.2642 3.913959 0.060137 0.988033 nosed at 6 and/or 12 months. A13-023-C 10885 366 425.7143 4.232292 0.035282 0.992926 Results: A total of 7 phyla and 399 fungal genera were distinguished A13-016-C 11250 573 778.8875 4.588823 0.026887 0.983822 in house dust. Fungal alpha diversity was similar between two groups. However, our results show that the different fungal communities C12-042-C 8323 341 639.5294 3.248262 0.102067 0.978974 between healthy and AD groups. The genera Alternaria, Aspergillus, A13-048-C-21 10255 504 647.3544 4.15016 0.049551 0.985275 Fusarium and Candida belonging to Ascomycota and known as the S12-056-C 11210 940 1505.511 4.965526 0.03022 0.964585 common allergen were higher abundant in dust samples for infants S12-052-C 10900 578 787 3.604678 0.118101 0.980734 with AD. On the other hand, Malassezia, Pleuotus and Trichosporon belonging to Basidiomycota were less abundantin dust samples for A12-051-C 11160 688 1004.469 4.548595 0.0311 0.977867 infants with AD. A12-036-C 11206 492 711.6133 3.816188 0.086481 0.983759 Conclusion: House dust from infants with AD show higher abundance C13-033-C 11982 562 863.8889 4.367747 0.030685 0.982557 of specific fungi known as allergens, which suggests that different Y13-018-C 11885 496 632.2805 4.114662 0.045829 0.987379 fungal exposure from the dust may link to the development of AD in infancy. A10-596-C 8604 438 726.0159 3.686982 0.075458 0.977801 Keywords: House Dust, Atopic Dermatitis, Infant, Fungal Communities S13-025-C 9653 503 786.5156 4.473125 0.025739 0.980213 S13-029-C 10485 580 815.6915 4.305426 0.03932 0.979876 P02 C14-006-C 11607 633 941.9326 4.389317 0.04185 0.979754 Cluster analysis in Bulgarian children with asthma 1 1 1 C14-009-C 10267 528 823.5882 4.297914 0.050246 0.980423 Snezhina Lazova , Penka Perenovska , Dimitrinka Miteva , Stamatios 2 1 Priftis , Guergana Petrova S12-090-C 10219 480 665.2442 4.017626 0.053966 0.982484 Medical University, Pediatric clinic, UMHAT Alexandrovska, Sofia, Bulgaria; S13-045-C 10677 421 560.2344 3.541324 0.118609 0.98745 Medical University, Sofia, Bulgaria Total 401206 20865 Correspondence: Snezhina Lazova - snejina@lazova.com Clinical and Translational Allergy 2017, 7(Suppl 2):P02 Clin Transl Allergy 2017, 7(Suppl 2):14 Page 6 of 17 Introduction: Asthma is a complex heterogeneous disease that likely Patients with both scoliosis and asthma, should be managed as severe comprises several distinct disease phenotypes. Usually atopy presence asthma patients. Evaluation of their PFT is essential in their manage- or any allergic diseases could interfere and obstacle the control of ment plan for early intervention should not only restriction type defi- asthma. Clustering approach has been used to classify heterogeneous ciency is noted asthma population into distinct phenotypes. Keywords: Scoliosis, Severe Asthma, FEV1 Method: For a period of 1  year we evaluated medical history data of 110 children with asthma aged 3 to 17  years. For all children we per- Acknowledgements: This work was supported by a grant from the formed pulmonary function tests, nasal smears for eosinophil counts, Medical University of Sofia (Council of Medical Science, project no. drew blood for IgE against inhalation and food allergies antibodies 513/2016, grant no. 65/2016). detection and ACQ. IgE were detected with the predesigned kit Euro- linePediatric and total IgE with Immunocap. Differences in clinical P04 indices including ΔFEV1(pre- and post-bronchodilator) and nasal and The cases of asthma in Japan have been expanding or not. Study blood biomarkers at enrollment among clusters were analyzed. on the tendency of asthma from the official statistical reports Results: Five distinct phenotypes were determined. Cluster 1 (n = 13): Takashi Kumae (non-atopic) the lowest IgE level, very low ACQ, median age of diag- Research Institute, Tokyo University of Agriculture, Tokyo, Japan nosis and IgE levels. Cluster 2 (n  =  45): (mixed) the highest BMI with Correspondence: Takashi Kumae - r071018@kanto-gakuin.ac.jp the latest age of diagnosis and high ACQ and BDR levels. Cluster 3 Clinical and Translational Allergy 2017, 7(Suppl 2):P04 (n = 33) (atopic) early diagnosis, highest BDR, highest ACQ score, high- est total and specific IgE levels among the clusters. Cluster 4 (n  = 12): Introduction: For consideration and discussion of data obtained from (atopic) the highest Immunocap result, relatedly high BMI and IgE with a survey on asthma, two official statistical reports are available in median ACQ score among clusters. Cluster 5 (n = 7): (non-atopic) the Japan. The fundamental data in these reports are statistically analyzed earliest age for diagnosis, with the lowest BMI, the lowest ACQ score, and compared to clarify the tendency of asthma in Japan. and Immunocap result, with high BDR and median level of IgE among Method: One is annual report of school health statistics (SHS) from clusters. Ministry of Education, Culture, Sports, Science and Technology which The patients with severe asthma were over 65% of the patients in clus- has competence to survey the school age population including dis- ter 3. Only one severe asthma patient was in cluster 2, and none in the eases and related health problems.  The incidence of asthma of SHS other three clusters. from 1995 to 2015 in 5 to 17 years old are used in this report. The other Conclusion: We identified asthma phenotypes in Bulgarian children is the patient survey by Ministry of health, Labour and Welfare. The according IgE levels, ACQ score, BDR and age of diagnosis. Almost all patient survey covers all generations and diseases but the survey car- of the severe asthma patients were in cluster 3 – with early diagnosis ried on every third year and numbers of patients are summarized by and highest IgE but not highest Immunocap results. 5 years old. Numbers of asthma case from 1996 to 2014 are extracted Keywords: Atopy, IgE, ACQ, Severe Asthma and adjusted to the corresponding age population as per 1,000 capita in this report. P03 Results: According to SHS, the incidence of asthma was most frequent Lung function parameters in asthma, severe asthma and scoliosis in 6  years old and statistically significant positive correlations were in children observed between the incidence and a lapse of years in all ages. Aver- 1 2 1 2 Snezhina Lazova, Vassil Yablanski , Penka P erenovska, Evgeni Vlaev , age slope of regression lines in the 5 to 9 years old was 0.10 in female 1 3 1 Hristina Rafailova , Stamatios Priftis , Guergana Petrova and 0.16 in male. This analysis means that the incidence may be keep Medical University, Pediatric cilinic, UMHAT Alexandrovska, Sofia, Bul- increase over 0.1% in annual in the near future. From the patient sur- 2 3 garia; Orthopedics department, Tokuda Hospital, Sofia, Bulgaria; Medical vey, asthma cases were most numerous in the 0 to 4  years old group University, Sofia, Bulgaria and the 5 to 9  years old group was next. Asthma cases increased in Correspondence: Snezhina Lazova - snejina@lazova.com elderly and the level of the 70 to 74 years old group was almost same Clinical and Translational Allergy 2017, 7(Suppl 2):P03 level of the 10 to 14 years old group. Contrary to SHS, statistically sig- nificant negative correlations were observed between the asthma cases and a lapse of years in the age groups, 15 to 19 (p  <  0.01), 20 Introduction: Pulmonary function testing (PFT ) is of great importance to 24 (p < 0.05) and 25 to 29 (p < 0.05) years old in male. The analysis in the evaluation of lung function. Spirometry is simple, noninvasive, of the patient survey suggests that the asthma has decreased in these and has been the most commonly used technique in children. Scolio- two decades. sis is the most common abnormality of the spine with direct effects Conclusion: Contradictory results in younger generation are obtained on the thoracic cage. Scoliosis has generally been associated with the by the analysis of the two official statistical reports. The patient sur - development of restrictive lung disease. vey is based on actual number of persons had a medical examina- Objectives: To evaluate the PFT data of children with scoliosis and to tion in medical facilities, on the other hand, SHS is carried out using compare them to children with asthma. the questionnaire. The data of SHS is seemed to be influenced by the Method: After obtaining signed informed consent from the parents anamnesis. Especially in lower age children, SHS data are also possibly we performed PFT in 50 children aged 5 to 17 years. The children were influenced by the levels of interests and anxieties for asthma in their divided into four groups – 10 children with scoliosis without asthma protectors. (Sc),10 children with scoliosis and asthma (AS), 10 children with mod- Keywords: Annual Report Of School Health Statistics, Patient Survey, erate asthma (BA), 10 children with severe asthma (SA) and 10 healthy Statistical Analysis children (HC). Results: The four groups have similar age (p  =  0.079), sex (p  =  0.19) P05 and FVC/FEV1 (p  =  0.403) distribution. The results for FVC  %pred, Patients’ perspectives of the impact of severe asthma on sex, FEV1%predand MMEF 25/75%pred were significantly different with intimacy and relationships: too taboo to talk about? p = 0.009 (Sc vs AS), p = 0.002 (SA vs BA) and p = 0.001 (AS vs BA), all 4 1 2 3 L. J. Holmes, J. Yorke , D. M. Ryan groups had lower results compared to HC (p < 0.05). The same param- University Hospital of South Manchester & University of Manchester, eters were similar when comparing Sc with BA and SA with AS. The Manchester, United Kingdom; University of Manchester, Manchester, lowest FVC %pred was in the AS group, even lower than SA (p = 0.06). United Kingdom; University Hospital of South Manchester, Manchester, The children with SA demonstrated the lowest MMEF 25/75%. The United Kingdom FEV1% pred were higher in healthy group while children with scoliosis Correspondence: Leanne-Jo Holmes - leanne.holmes1@ntlworld.com and asthma have comparable lower values. Clinical and Translational Allergy 2017, 7(Suppl 2):P05 Conclusion: The children with scoliosis demonstrated diminished expiratory flow rates, while the FEV1/FVC ratio is within normal ranges. Clin Transl Allergy 2017, 7(Suppl 2):14 Page 7 of 17 Introduction: Patients with severe asthma experience unpredictable measured by ELISA. Spot urine samples were measured for urinary daily symptoms and an intense treatment regimen that can impact Leukotriene E4 (uLTE4) levels by enzyme immunoassay. upon physical, emotional well-being and quality of life. Sexual func- Results: Two hundred and nine data collecting forms with the bio- tion is a basic human requirement, contributing to quality of life; yet assay results were available for analysis in this report. The mean age there is a dearth of research exploring relationships and intimacy in of the participants was 26  months with boys making up 62.5%. First people with severe asthma. The overall aim was to explore the impact wheeze episode accounted for 40.6 percent. Seventy-five percent was of severe asthma on patient’s experiences of intimacy and relation- atopic with mono-sensitization (29.3%). The highest prevalence of ships, establish their information needs and the role of the healthcare sensitization was egg (45.6%), followed by cow’s milk (45.3%). The high professional. median urine leukotriene E4 (uLTE4) levels of 226  pg/ml and 956  pg/ Method: A qualitative study using semi-structured interviews with mg creatinine was documented. The prevalence of vitamin D insuffi- patients receiving treatment for severe asthma at a dedicated severe ciency, defined by the serum 25OHD levels of less than 30 ng/ml, was asthma clinic was undertaken. Interviews were audio recorded and detected in 22.3%. transcribed verbatim. Transcripts were verified then analysed for emer - Conclusion: Majority of preschool children with acute wheezing gent themes which were then categorised into super-ordinate themes admitted in the hospital had sensitization with egg and cow’s milk. and sub themes. One-fourth of them had vitamin D insufficiency and associated with Results: All nine participants had a proven diagnosis of severe high serum level of uLTE4. asthma. A higher proportion were female n  =  6 (66.6%) compared Keywords: Allergic Sensitization, Acute Wheezing to male n = 3(33.3%). The mean age was 45 years, range 34–59 years. The majority of participants 8(88.9%) were married, with 1(11.1%) P07 co-habiting. Cluster analysis of patients with moderate to severe uncontrolled The analysis of interviews provided complex, detailed and novel bronchial asthma insights into patients’ perspectives on how living with severe asthma 1 1 1 Vania Youroukova , Denitsa Dimitrova, Yanina Slavova , Spaska impacts upon intimacy and spousal relationships. Four super-ordinate 2 Lesichkova themes emerged: 1 Medical Faculty of Medical University of Sofia; Clinical center of pulmo - nary diseases; SHATPD “St. Sofia”, Sofia, Bulgaria; Medical Faculty of Medi- cal University of Sofia, Department of Clinical Immunology, University (i) ‘Physical intimacy’ including disclosure of physical limitations of Hospital Alexandrovska, Sofia, Bulgaria severe asthma upon intimacy and sexual activity. Correspondence: Denitsa Dimitrova - ddimitrova87@abv.bg (ii) ‘Emotional intimacy’ the cyclical impact of the often negative Clinical and Translational Allergy 2017, 7(Suppl 2):P07 emotional struggle of living with severe asthma upon both rela- tionships and intimacy. (iii) ‘Image of self’ as participants divulged their battle with body Introduction: The aim of the study is to identify distinct subgroups of image and confusion in changing relationship roles. adult patients with uncontrolled moderate to severe bronchial asthma (iv) ‘The role of the healthcare professional’ patient identified infor - (BA) based on clinical and inflammatory characteristics. mation requirements within this area. Method: Adults with uncontrolled moderate to severe asthma (n  =  23) underwent a clinical assessment, spirometry and measure- Conclusion: The participants in this study have provided invaluable ment of fractional exhaled nitric oxide (FeNO). Total and specific serum and novel insights into the implications of living with severe asthma IgE levels, eosinophil count in peripheral blood and sputum were eval- upon a sensitive and rarely discussed topic. It is anticipated that these uated. K-mean cluster analysis was used to identify clinically relevant findings will serve to increase understanding and assist practitioners subgroups. to help patients to adopt positive strategies to improve their quality of Results: We have identified four distinct groups based on onset, atopic life within this area. status, FEV1/FVC ratio and FeNO. Cluster 1 patients (eosinophilic asthma) (n  =  6; 26.1%) were with late-onset asthma, predominantly non-atopic with deteriorated lung function and highest level of FeNO. P06 In this cluster were observed the highest blood and sputum eosino- Clinical characteristics and allergic sensitization of preschool phil counts (p = 0.01; p = 0.04) and more frequent exacerbations (≥ 1 children hospitalized with acute wheezing: a multi‑cohort study 1 2 exacerbation in previous 12 months) (p = 0.03). Cluster 2 patients (late- Sasawan Chinratanapisit , Khlongtip M atchimmadamrong , Jitladda 3 4 5 onset, atopic asthma) (n  =  7; 30.4%) were late-onset, mainly atopic Deerojanawong, Wissaroot Karoonboonyanan , Paskorn Sritipsukho with high total serum IgE levels, lower levels of FeNO and eosinophil Division of Allergy & Immunology, Department of Pediatrics, Bhumi- counts and deteriorated lung function. Cluster 3 patients (non-atopic bol Adulyadej Hospital, Bangkok, Thailand; Saraburi hospital, Saraburi, asthma) (n = 6; 26.1%) were late-onset, mainly non-atopic, with lower Thailand; Division of Respiratory disease and intensive care, Department levels of FeNO and eosinophil counts and well-preserved pulmonary of Pediatrics, Faculty of Medicine, Chulalongkorn University, Bangkok, function. Cluster 4 patients (early-onset, atopic asthma) (n = 4; 17.4%) Thailand; Department of Pediatrics, Bhumibol Adulyadej Hospital, Bang- were early-onset, atopic, with low levels of FeNO and preserved lung kok, Thailand; Center of Excellence in Applied Epidemiology, Thammasat function. In cluster 3 and 4 prevailed patients with high BMI (n = 7). University, Bangkok, Thailand Conclusion: Our current results suggests that uncontrolled asthma is Correspondence: Sasawan Chinratanapisit - sasawan2001@yahoo.com heterogeneous disease. Cluster identification will result in a personal- Clinical and Translational Allergy 2017, 7(Suppl 2):P06 ized approach for each patient and improvement of therapy response. The research was funded by GRANT project № 308/2015, contract Introduction: Background: Wheezing is a significant health problem №75/2015 to CMS, MU Sofia in Thailand, especially in the preschool age. Keywords: Clusters, Uncontrolled Adult Asthma, Inflammatory Objective: In this multi-center clinical research, clinical characteristics Characteristics and biomarkers were studied to determine the factors associated with persistent wheezing in the next three years among pediatric patients P08 with acute wheezing who were less than five years of age. Influence of respiratory viruses on the severity of bronchial Method: This prospective observational study recruited children, aged obstruction in preschool wheezing children 6 months to 5 years, with acute wheezing from the pediatric wards of 1 1 1 2 Iren Tzocheva , Snezhina Lazova , Snezhana Parina , Svetla Angelova , four hospitals from July 2014 to June 2015. Patients’ characteristics 2 1 Neli Korsun , Penka Perenovska and their family data were collected. Blood samples were collected for Pediatric clinic, University Hospital Alexandrovska, Medical University, detecting ImmunoCAP specific IgE against common food and inhal- Sofia, Bulgaria; National Centre of Infectious and Parasitic Diseases, ant allergens. Serum 25-hydroxyvitamin D (25OHD) concentration was Clin Transl Allergy 2017, 7(Suppl 2):14 Page 8 of 17 National Laboratory Influenza and ARD, Sofia, Bulgaria Results: In 2014 were 880 ED visits of 296 patients. 16 children were Correspondence: Snezhina Lazova - snejina@lazova.com included in the tele-monitoring study, 4 with severe asthma and 38 as Clinical and Translational Allergy 2017, 7(Suppl 2):P08 controls. Patients had male predominance [75% vs 71.1% in controls]. 2 (1 of the 4 severe asthma patients) vs 11 children in control group had more than 1 visit to ED in 2014. Odds-ratio to have an unsched- Introduction: Respiratory infections are associated with wheezing ill- uled ED visit was 0.1964 in tele-monitoring group [p 0.0483, 95% CI nesses at all ages and may also impact the development and sever- 0.039 – 0.9881]. Was higher for severe asthma patients compared with ity of asthma. Wheezing illnesses in young children are associated up non-severe with an odds–ratio 3.667 but non-significant [p 0.4040, to 95% with respiratory viral infections. In children with established 95% CI 0.1734 to 77.5559] asthma, viral respiratory tract infections play a key role in producing Conclusion: Tele-monitoring via virtual media can decrease the bur- acute exacerbations that may lead to hospital admission. The aim of den of unscheduled ED visits for asthmatic children, even in severe the present study is to investigate the relationship between various asthma patients. respiratory pathogen and severity of bronchial obstruction in hospital- Keywords: Child, Asthma, Virtual-Media, Monitoring, Education ized preschool children. Method: Nasopharyngeal secretions of 57 children aged 2 to 5  years P10 hospitalized with wheezing episode (Jan to Mar 2016) were examined Challenges in using hierarchical clustering to identify asthma for respiratory viruses (HMPV; RSV A/B; PIV 1, 2, 3; RV, AdV; Influenza subtypes: choosing the variables and variable transformation A/B) with Real Time PCR. 1 2 3 4 Matea Deliu, Tolga Yavuz , Matthew Sperrin , Umit M. Sahiner , Danielle Results: Respiratory viral infections was detected in 84% of the 5 6 7 8 Belgrave , Cansin Sackesen Sackesen , Adnan C ustovic , Ömer Kalayci patients. The predominant pathogen were RSV (52%), followed by Institute of Population Health, Health e-Research Centre, University influenza virus A (25%), human metapneumovirus (12.5%), rhinovirus of Manchester, Manchester, United Kingdom; Division of Pediatric (4%), parainfluenza (4%) and adenovirus (2.5%). In children with life- Allergy, GATA Military School of Medicine, Ankara, Turkey; Centre threatening bronchial obstruction with a need of prolonged oxygen for Health Informatics, Institute of Population Health, University of Man- therapy, RSV was the most commonly detected pathogen (42%). The chester, Manchester, United Kingdom; Division of Pediatric Allergy, presence of viral infection did not correlate with the duration of the Hacettepe University School of Medicine, Ankara, Turkey; Faculty systemic corticosteroid treatment. The median duration of reported of Medicine, Department of Medicine, Imperial College London, London, lung sounds (wheezing and crackles) was longer in the patients with 6 7 United Kingdom; Koç University Hospital, Istanbul, Turkey; Department detected virus infection without statistical significance (p = 0.06). of Paediatrics, Imperial College London, London, United Kingdom; Pedi- Conclusion: Respiratory viruses play a key role in the wheez- atric Allergy and Asthma Unit, Hacettepe University School of Medicine, ing illnesses in young children, predominantly infants. Our results Ankara, Turkey demonstrated that the RSV is an important pathogen in severe, life- Correspondence: Matea Deliu - matea.deliu-2@postgrad.manchester. threatening episodes in preschool age children as well. Contrary to ac.uk data presented in literature, human rhinovirus was detected in only 4% Clinical and Translational Allergy 2017, 7(Suppl 2):P10 of our patients following Influenza A and human metapneumovirus. Keywords: Preschool Wheezing, Respiratory Viruses, Asthma Exacer- bation, Severity Introduction: The use of unsupervised clustering has identified differ - ent subtypes of asthma. Choosing the variables to input into the clus- P09 tering algorithm is one of the important considerations. The majority Tele‑monitoring decreases unscheduled outpatient visits of previous studies selected variables based on expert advice, whilst in pediatric patients with severe asthma others used dimension reduction techniques such as principal compo- Mihai Craiu, Iustina Violeta Stan nent analysis (PCA). We aimed to compare the results of unsupervised INSMC Alessandrescu-Rusescu, Bucharest, Romania clustering when using raw variables, or variables transformed using Correspondence: Mihai Craiu - mcraiu@yahoo.com dimensionality reduction techniques. Clinical and Translational Allergy 2017, 7(Suppl 2):P09 Method: We performed our analysis on 613 asthmatics aged 6–23 years from Ankara, Turkey. We conducted extensive phenotyping and recorded 49 variables including demographic data, sensitization, Introduction: Pediatric asthma patients originating from poor- lung function, medication, peripheral eosinophilia, and markers of resource areas experience a significant increase of unscheduled and asthma severity. We performed hierarchical clustering (HC) using: (1) emergency department visits and most of medical care for them is all variables; and (2) variables transformed using dimensionality reduc- delivered in such a facility. Education and communication is vital for tion techniques. severe asthma patients. Virtual media can be used to improve asthma Results: PCA revealed 5 components describing atopy and variations control in these patients. Official data are documenting that ¾ of citi- in asthma severity, which were then used to infer cluster assignment. zens own at least a smart-phone in our country and even in poor areas The optimal HC solution in both PCA-transformed and raw untrans- there is excellent mobile phone coverage. The aim of our study was to formed data identified five clusters. However, these clusters were not document outcome of a structured tele-monitoring approach (via vir- identical. Both identified mild asthma with good lung function, severe tual media) in management of severe asthmatic children. atopic asthma and late-onset mild atopic asthma. However, the over- Method: Prospective 12  months study of patients with severe asthma. lap between children assigned to these three clusters in two HC analy- Were observed severe asthma patients from a cohort of previously mon- ses was modest. Clustering without PCA identified early-onset severe itored children with uncontrolled asthma [ACT child score below 19] atopic asthma and late-onset atopic asthma with high BMI, whilst and frequent visits in Emergency Department in previous year. Exten- early onset non-atopic mild asthma in females was identified in HC sive education in basic inhalation techniques [video-training] was done with PCA. and written-action plan (WAP) was provided. Were trained to use PIS Conclusion: Different methodologies applied to the same dataset [Pulmonary Index Score] to evaluate severity of exacerbation and treat- identified differing clusters of asthma. Despite cluster instability, both ment response. Parents had to either use ED or use WAP and tele-mon- methodologies provided meaningful clinical insights for understand- itoring. Short video of clinical status was evaluated by an expert and ing asthma heterogeneity. further options were discussed. Non-responders were presented in ED. Clin Transl Allergy 2017, 7(Suppl 2):14 Page 9 of 17 POSTER DISCUSSION SESSION 1—Biomarkers generated sequences and/or number of samples. A correlation matrix was used to identify positive and negative correlations between bac- terial and viral families and the results were superimposed in a net- P11 work graph. Th17 in children with asthma 1 1 1 Results: Taxon classification revealed 198 bacterial and 20 viral fami- Snezhina Lazova , Penka Perenovska , Guergana Petrova , Dimitrinka 1 2 3 3 lies. Unsupervised hierarchical clustering based on beta diversity (Jac- Miteva, Petar Velikov, Tsvetelina Velikova , Ekaterina Ivanova-Todorova , 3 3 card matrix) clustered controlled, partly controlled and uncontrolled Kalina Tumangelova-Yuzeir , Dobroslav Kyurkchiev 1 asthma separately. Topographical analysis of the metagenomic inter- Medical University, Pediatric clinic, UMHAT Alexandrovska, Sofia, Bulgaria; 2 3 action map identified the phage family of Myoviridae as a central Medical University, Sofia, Bulgaria; Medical University, Laboratory component. Further analysis of the Myoviridae family and its direct of Clinical Immunology, University Hospital St. Ivan Rilski, Sofia, Bulgaria interactions with bacterial families (17 families) revealed that patients Correspondence: Snezhina Lazova - snejina@lazova.com with partly controlled and uncontrolled asthma have reduced quantity Clinical and Translational Allergy 2017, 7(Suppl 2):P11 of Myoviridae phages and reduced diversity and quantity of their bac- terial interactors. Introduction: Th17 lymphocytes are now widely believed to be criti- Conclusion: Based on the Rarefaction curves, we estimate that, even cal for the regulation of various chronic immune diseases, including with 5 samples a high percentage of the microbial diversity can be asthma and chronic obstructive pulmonary disease. robustly represented. Asthma patients were efficiently clustered Objectives: To assess Th17 levels in children with asthma (BA) and based on their microbial similarity which seems to be representative cystic fibrosis (CF). of asthma control. The phage family Myoviridae is predicted to hold Method: We included 12 children with CF without history of allergies a central role in the viral-bacterial interaction on the specific environ- and 20 children with BA aged 7 to 16 years (10 with severe BA and 10 mental site. The reduced number of reads of the Myoviridae phages with moderate BA). After informed consent, 1.2  ml of venous blood along with the reduced diversity and quantity of 17 bacterial families was collected during a routinely performed blood withdrawal. Analy- in uncontrolled asthma could be evident of an ongoing ‘fight’ between ses of Th17 cells were performed with Lise-Wash Protocol by a 4-color components of the upper respiratory metagenome. FacsCalibur flow cytometer after staining the whole blood with the Keywords: Asthma, Metagenomics, Respiratory, Children following fluorescence-labeled antibodies: anti-CD3 (FITC), anti-CD (PerCP), anti-CD161 (PE) and anti-CCR6 (AlexaFluor 647). P13 Results: The patients in asthma group had significantly higher per - Circadian breathomics in asthma: analysis by thermal centage of Th17 12.40%  ±  1.16 compared to the children with CF - desorption‑gas chromatography‑mass spectrometry 7.64 ± 0.87 (p = 0.008). Stratifying the BA group according the severity 1 1 2 3 Maxim Wilkinson , Craig P ortsmouth , David Ray , Royston Goodacre , we found higher levels of Th17 in patients with severe BA (p  <  0.05). 4 2 Stephen J. Fowler , Hannah Durrington Patients with moderate asthma had TH17 values close to those in CF 1 Division of Infection, Immunity and Respiratory Medicine, Faculty of Biol- children. ogy, Medicine and Health, University of Manchester, Manchester, United Conclusion: The number of Th17 cells is significantly increased in the 2 Kingdom; Division of Metabolism and Gastroenterology, Faculty of Biol- peripheral blood of children with severe BA compared to the children ogy, Medicine and Health, University of Manchester, Manchester, United with moderate BA. Severe BA patients could have possible benefit 3 Kingdom; School of Chemistry, Manchester Institute of Biotechnology, from the future target therapies. 4 University of Manchester, Manchester, United Kingdom; Manchester Keywords: Severe Asthma, Controlled Asthma, Cystic Fibrosis Academic Health Science Centre, and NIHR Respiratory and Allergy Clinical Research Facility, University of Manchester, University Hospital of South Manchester, Manchester, United Kingdom Acknowledgements: This work was supported by a grant from the Correspondence: Maxim Wilkinson - maxim.wilkinson@postgrad. Medical University of Sofia (Council of Medical Science, project no. manchester.ac.uk 289/2015, grant no. 54/2015). Clinical and Translational Allergy 2017, 7(Suppl 2):P13 P12 Study of the viral and bacterial communities associated Introduction: Asthma is closely associated with the endogenous cir- with asthma: a metagenomic approach cadian rhythm of the lungs; in severe and uncontrolled asthma noctur- 1 1 Spyridon Megremis , Bede Constantinides , Alexandros Georgios nal wakening is common, as is a dip in early morning peak expiratory 1 2 1 Sotiropoulos , Paraskevi Xepapadaki , David Robertson , Nikolaos flow. This variability makes asthma a potential target for chronother - Papadopoulos apy to improve the efficacy of treatment. To personalise our approach 1 2 University of Manchester, Manchester, United Kingdom; University to chronotherapy in asthma, it is essential to have an easy-to-meas- of Athens, Athens, Greece ure biomarker that reflects an individual patient’s chronotype. This Correspondence: Spyridon Megremis - spyridon.megremis@manches- would allow chronotherapy to be targeted to the right time of day ter.ac.uk for each patient. Breath samples are easy to collect and non-invasive, Clinical and Translational Allergy 2017, 7(Suppl 2):P12 and exhaled volatile organic compounds (VOCs) have been related to airway inflammatory patterns in asthma. We hypothesise that the exhaled VOC profile contains potential biomarkers that track the cir - Introduction: An important unexplored source of interpersonal vari- cadian rhythm in asthma. Samples are analysed using thermal desorp- ation in asthma presentation is the complex and dynamic commu- tion-gas chromatography–mass spectrometry which is currently the nity of microorganisms located on different sites of the respiratory gold standard for offline VOC detection, and allows high sensitivity tract. Our aim was to identify and explore the bacterial and viral diver- and reliable identification of detected compounds. sity in a small pilot study of asthmatic children using metagenomics. Method: Twenty non-smoking volunteers (10 healthy, 10 with mod- Method: Nasopharyngeal samples were obtained from five asth- erate asthma) will be recruited. So far nine individuals have been matic children (2 males) at a baseline asymptomatic visit. Mean age recruited; the study design is detailed in the figure. Breath samples are was 5.2  years (range 4.6–5.8). Communities of viruses (DNA and RNA taken at 15:00 on the first visit; 15:00, 21:00, 03:00 and 09:00 on the genomes) and bacteria were evaluated using high depth paired- second and 09:00 on the final visit. This gives six samples per partici- end shotgun sequencing. We performed taxonomic classification of pant at four time points allowing investigation of both the changes in sequences and calculated environmental indexes (alpha and beta) to an individual’s VOC profiles over the course of a day and longitudinal estimate the microbial diversity both within (Shannon, Chao1, Jack1) variation over the course of several weeks. All samples are analysed and between (Jaccard) patients. Rarefaction curves were produced to using thermal desorption-gas chromatography-mass spectrometry study the relationship between microbial diversity and the number of Clin Transl Allergy 2017, 7(Suppl 2):14 Page 10 of 17 Results: The extracted ion chromatograms for the internal standard low dose ICS (n  =  11) therapy (p  =  0.011, p  =  0.007 respectively). (p-bromofluorobenzene), isoprene, limonene and the alkanes are Twenty AP (n  =  5, CA and n  =  15, UA) demonstrated impaired lung shown as a quality control procedure to ensure clean data process-function (FEV   <  80%), which was associated with higher levels of SP ing. Preliminary results will be presented by multivariate analysis for (p = 0.04). Similarly, in UA was observed significantly negative correla- the first nine participants comprising six individuals with asthma and tion between SP and FEV  % (p = 0.006, r = − 0.56). three controls. Conclusion: Patients with persistent moderate to severe bronchial Conclusion: Measuring exhaled breath VOCs at intervals through- asthma show higher levels of SP as compared to healthy controls. High out the day may provide a useful way of mapping a patient’s asthma levels of SP are associated with patient need of higher dose ICS ther- chronotype, guiding future chronotherapy. apy. Our results suggest that SP is biomarker for impaired lung func- Keywords: Circadian Rhythm, TD-GC–MS, Breathomics tion in poor asthma control. Keywords: Periostin, Severe Adult Asthma, Lung Function The research was funded by GRANT project No. 308/2015, contract No. 75/2015 to CMS, MU Sofia P15 Serum periostin values in healthy non‑asthma non‑copd spanish adults Irina Bobolea, Daiana Guillén Vera, Gabriel Gonzalez-Salazar, Carlos Melero Moreno, Consuelo Fernandez Rodriguez, Natividad De Las Cuevas Moreno Hospital 12 de octubre, i + 12 Research Institute, Madrid, Spain Correspondence: Irina Bobolea - ibobolea@gmail.com Clinical and Translational Allergy 2017, 7(Suppl 2):P15 Introduction: Serum periostin is an emerging biomarker for T2 asthma, and for the response to new biologicals such as anti IL-13 lebrikizumab. But a clear cut-off has not been established, and other pathologies beyond asthma are known to course with high periostin levels. The aim of this study was to establish reference values for serum periostin in a representative sample of non-asthmatic non-COPD Figure 4 Schematic of the experimental design healthy population in our geographical area. Method: A cross-sectional study was conducted between May–Octo- ber 2015. After signing the informed consent, we collected sera sam- ples from consecutive blood donors of the Blood Bank of Universitary Hospital 12 de Octubre, Madrid, Spain. By choosing this source of donors, all diseases and other conditions P14 related to any periostin elevations described so far were excluded, as Serum periostin levels as a biomarker for impaired lung function they constitute contraindications for donating blood: chronic illnesses in adult patients with moderate to severe asthma 1 1 2 (diabetes; heart, pulmonary, renal or hepatic diseases); any type of Denitsa Dimitrova, Vania Youroukova, Tsvetelina Velikova , Kalina 2 3 cancer; chronic or acute infectious diseases, including stomathology- Tumangelova-Yuzeir, Anna Valerieva 1 cal disordes; pregnancy; recent traumatic injury or surgery. Moreover, Medical Faculty of Medical University of Sofia; Clinical center of pulmo - 2 all participants completed a questionnaire aimed to exclude those nary diseases; SHATPD “St. Sofia”, Sofia, Bulgaria; Medical Faculty of Medi- with symptoms and/or a physician diagnosis of asthma and/or COPD. cal University of Sofia, Department of Clinical Laboratory and Clinical 3 Sera were frozen at −80 °C until periostin was measured using an EISA Immunology, University Hospital, Sofia, Bulgaria; Medical Faculty commercial kit. We calculated the sample size to estimate a media, of Medical University of Sofia, Clinic of Allergy and Asthma, University using a standard deviation (SD) of 15, with 3 ng/mL precision and 95% Hospital Alexandrovska, Sofia, Bulgaria confidence interval (Anastasilakis et al. 2014). Correspondence: Denitsa Dimitrova - ddimitrova87@abv.bg Results: 100 non-asthmatic non-COPD and otherwise healthy sub- Clinical and Translational Allergy 2017, 7(Suppl 2):P14 jects, aged between 18 and 65  years, were included in the study. Results are expressed in media (SD) (min–max range). Age: 43.97 years Introduction: The aim of the study is to determine the relationship (10.35) (21–65  years). 62 males (62%). 22 smokers (25.9%); being the between serum periostin (SP) levels and clinical characteristics in adult official percentage of smokers in the Madrid area 27.4%. patients with moderate to severe asthma (AP). Serum periostin: 33.04  ng/mL (14.37) (6.87–81.43  ng/mL). The media Method: SP levels were measured by ELISA method in 42 AP out of estimated in total population was (30.19–35.90  ng/mL) with a con- exacerbations and 10 healthy controls (HC). AP were divided into two fidence interval of 95%. Periostin in males: 33.51  ng/mL (14.02), and groups according asthma control test (ACT) score: controlled AP (CA, females: 32.28 ng/mL (15.09), p = 0.680. Smokers: 32.4 ng/mL (12.05) n = 19) and uncontrolled AP (UA, n = 23). Atopic status was confirmed and non-smokers: 33.09  ng/mL (15.28) respectively, p  =  0.850. No by skin prick testing and measurement of total and specific serum IgE association was either found between age and serum periostin levels. Lung functional parameters were assessed by spirometry. (r = 0.075). Results: We have observed significantly higher levels of SP in AP Conclusion: We report reference values for serum periostin in a (1.68  ±  0.52  ng/ml) compared to HC (1.08  ±  0.12  ng/ml) (p  <  0.001). healthy non-asthmatic non-COPD adult Spanish population. Our No statistically significant differences were found in SP levels between results could be used as reference for further studies aimed to clarify CA and UA, atopic (n = 14) and non-atopic AP (n = 28), current smok- the role of periostin in asthma phenotypes and endotypes, or to ers (n  =  13) and non-smokers (n  =  29) (p  =  0.85, p  =  0.33, p  =  0.19 choose future candidates for the new biological agents targeting T2 respectively). We have found significantly higher SP levels in AP on inflammation. medium (n  =  22) and high doses ICS (n  =  9) compared to those on Keywords: Asthma Biomarkers, Periostin Clin Transl Allergy 2017, 7(Suppl 2):14 Page 11 of 17 POSTER DISCUSSION SESSION 2—Mechanisms cells and is involved in lymphocyte activation, proliferation and differ - entiation. Targeting PI3  Kδ may have therapeutic benefit in currently uncontrolled asthma patients. P18 The aims of the study were to compare the expression PI3 Kδ in airway ToAST: Investigating the effects of bronchial thermoplasty (bt) samples from asthma patients and healthy subjects and to assess the on cough responses in patients with severe asthma – a pilot study anti-inflammatory potential of PI3  Kδ inhibitors on bronchoalveolar R. Wang, I. Satia, R. Niven, S. J. Fowler, D. M. Ryan, J. A. Smith lavage (BAL) lymphocytes. University Hospital of South Manchester, Manchester, United Kingdom Method: Bronchial biopsies, BAL cells and blood cells were collected Correspondence: Ran Wang - ranwang1986@googlemail.com from moderate to severe asthma patients and healthy subjects. The Clinical and Translational Allergy 2017, 7(Suppl 2):P18 numbers of PI3  Kδ-positive cells in biopsies was calculated using immunohistochemistry and the expression of PI3  Kδ in BAL CD3 cells Introduction: Severe asthma affects 5% of the asthma population but was assessed by flow cytometry. BAL and blood cells were treated with drives the majority of the morbidity and cost. Despite cough being a a PI3 Kδ inhibitor and/or dexamethasone prior to T cell receptor (TCR) major symptom and a marker of both severity and poor prognosis, stimulation. ELISAs were used to assess levels of IFNγ, IL-13 and IL-17 current medications are not designed to treat cough directly. Bron- and flow cytometry was used to measure pSTAT5, an early marker of chial thermoplasty (BT) is a novel therapy in severe asthma, deliver- T-cell activation. ing radiofrequency thermal energy to the large airways. There are 1 Results: Bronchial biopsies from asthma patients contained increased provisional data suggesting BT reduces the number of airway nerves . numbers of PI3 Kδ-positive cells compared to those from healthy sub- If this is the case, then BT may affect cough in asthma. Inhaled cap - jects, and asthma BAL T-cells expressed higher levels of PI3  Kδ than saicin challenge has been safely used to evoke cough responses and healthy cells. Inhibition of PI3  Kδ reduced TCR-stimulated IFNγ, IL-13 to assess cough threshold in research. However, to date no study has and IL-17 in BAL cells from both asthma patients and healthy subjects, adopted this method to assess cough in patients with severe asthma and treating BAL cells with a combination of PI3 Kδ inhibitor and dexa- or following BT. methasone had an additive effect on cytokine inhibition. TCR-induced We aim to assess the safety and feasibility of capsaicin challenge in pSTAT5 in purified T-cells and BAL cells was impeded by PI3 Kδ inhibi- people with severe asthma. We will also explore the effects of BT on tion and dexamethasone. 24 h cough frequency, capsaicin evoked cough responses and cough- Conclusion: Inhibition of PI3 Kδ in asthma may be an effective method related quality of life. of reducing activation of T-cells and subsequent cytokine production. Method: The protocol for this two-visit observational study has been Keywords: Phosphatidylinositol 3-Kinase Delta, Airway T-Cells, Bron- approved by the local Research Ethic Committee. All recruited patients choscopy, Corticosteroids, Cytokines (target n = 24, of whom half will have had BT ) will be treated at British Thoracic Society steps 4 or 5. During visit 1 patients will be consented, the Leicester Cough and Asthma Control Questionnaires administered P20 and baseline spirometry performed. They will undergo ambulatory Investigating neuronal responses by assessing capsaicin evoked 24-hour cough monitoring. A capsaicin challenge will be performed cough responses in an allergen challenge model of asthma (INCA) 1 1 1 2 during the second visit. Spirometry will be performed before and Imran Satia , Stephen Fowler, Mark Woodhead , Paul O’Byrne , Jaclyn immediately after the challenge to assess any change in FEV .Ann Smith 1 2 Results: The primary outcome measure will be the number of participants University of Manchester, Manchester, United Kingdom; McMaster with a 20% or greater fall in FEV1 and/or requiring rescue bronchodila- University, Hamilton, Canada tor therapy during the capsaicin challenge period. Secondary outcomes Correspondence: Imran Satia - imran.satia@manchester.ac.uk include: change in   %FEV1 predicted during capsaicin challenge; toler- Clinical and Translational Allergy 2017, 7(Suppl 2):P20 ability of inhaled capsaicin; number of adverse events during and after capsaicin evoked cough challenge; differences in capsaicin dose response Introduction: Cough is a common and troublesome symptom in curves between patients who have had BT versus those have not. asthma but little is known about the neuronal pathways that trigger Conclusion: This pilot study will lead to a larger prospective study cough. The mechanisms by which  airway inflammation, bronchial investigating the effects of BT on cough responses in patients with hyper-responsiveness and variable airflow obstruction cause cough severe asthma. is unclear. We have previously shown that methacholine induced bronchoconstriction heightens cough responses to inhaled capsaicin. Reference The aim of the current study was to investigate the effects of allergen 1. Bergqvist A, Pretolani M, Taille C, Dombret MC, Knapp D, Bjermer L, exposure on cough reflex sensitivity. Chanez P, Erjefalt JS, Aubier M. Selective structural changes of bronchial Method: We performed a 8 visit randomised, single-blind, placebo thermoplasty in the treatment of severe uncontrolled asthma. Am J controlled, two-way cross-over study comparing cough responses Respir Crit Care Med. 2015;191:A4171. to inhaled capsaicin in allergic asthma patients during and 24  h after exposure to allergen compared with diluent (saline) control. P19 Mild atopic asthmatics who are dual responders will undergo full dose Anti‑inflammatory capability of Phosphatidylinositol 3‑kinase allergen/diluent (saline) challenge. Early asthmatic responses (EAR) delta inhibitors in moderate to severe asthma patients (fall in FEV1 > 20% at 30 min) and late asthmatic responses (LAR) (fall 1 1 1 1 2 2 T. Southworth , J. Plumb, V. Gupta , J. P earson , I. Ramis , M. D. Lehner , in FEV1 > 15% at 3–7 h) will be documented. Full dose capsaicin chal- 2 1 M. Miralpeix , D. Singh lenge was performed at screening to determine the excitatory dose 1 2 The University of Manchester, Manchester, United Kingdom; Almirall S. evoking half the maximum cough response (ED50). The ED50 concen- A., Barcelona, Spain tration of capsaicin was inhaled 4 times, 30 s apart, at 30 min and 24 h Correspondence: Thomas Southworth - tsouthworth@meu.org.uk after inhaled allergen/diluent challenge. Cough counts will be meas- Clinical and Translational Allergy 2017, 7(Suppl 2):P19 ured throughout, along with methacholine challenge and induced sputum before and after allergen challenge. Introduction: Lymphocytes numbers are increased in the lungs of Results: We report interim analysis of the first 5 completed patients asthma patients, with T-helper 2 cells being associated with eosino- recruited to this study (mean age (SD) 23.4 yrs (4.6), BMI 23.9 (4.3), philic inflammation. There is also growing evidence that T-helper 1 FEV1% predicted 97.4% (8.0), PC20 1.55 (1.72), 2 female). All patients and T-helper 17 cells may play a role in more severe asthma. Inhaled were steroid naive and sensitive to house dust mites. Mean fall in corticosteroids (ICS) are widely used anti-inflammatory treatments for %FEV1 from baseline in the EAR was 34.9% (SD6.4), and LAR 29.2% asthma, but many moderate to severe asthma patients suffer from per - (20.6). Baseline ED50 concentration was 24.2microM(23.8). There was sistent symptoms, despite using high doses of ICS. Phosphatidylinosi- an increase in capsaicin evoked coughs after allergen exposure com- tol 3-kinase delta (PI3 Kδ) is predominantly expressed in inflammatory pared to diluent at both 30 min and 24 h (mean coughs after allergen Clin Transl Allergy 2017, 7(Suppl 2):14 Page 12 of 17 work is addressing the ability of Af activated DCs to influence T cell activation and polarization in vitro and in vivo. Keywords: Dendritic Cells, Bronchial Epithelial Cells, Aspergillus Fumigatus P22 Flavonoid cyanidin acts through IL‑17RA to alleviate IL‑17A‑mediated severe asthma Caini Liu, Liang Zhu, Kiochi Fukuda, Cunjin Zhang, Suidong Ouyang, Xing Chen, Luke Qin, Suguna Rachakonda, Mark Aronica, Jun Qin, Xiaoxia Li Figure 5 FEV1 and ED 50 Coughs 30 min and 24 h after Allergen/Diluent Cleveland Clinic, Cleveland, United States Correspondence: Caini Liu - liuc@ccf.org Clinical and Translational Allergy 2017, 7(Suppl 2):P22 vs. diluent at 30 min 15.0 (4.9) vs. 9.6(3.5), at 24 h 12.2 (8.0) vs. 6 (2.6). Introduction: Interleukin (IL)-17A plays a critical role in the pathogen- Figure 1 esis of asthma. High levels of IL-17A are found in bronchial biopsies Conclusion: We have safely performed capsaicin challenge after aller- and serum obtained from patients with severe asthma. Deficiency of gen exposure. Initial data suggests cough responses may be height- IL-17A signaling components attenuated airway inflammation in sev - ened for up to 24 h after allergen exposure. eral mouse models of asthma. We recently identified a potent small Our results show the importance of STAT6 and the possible implica- molecule compound A18 (called cyanidin, a key pigment present in tion of SOCS proteins in the enhanced CCL26 expression by BECs red berries and other fruits) for IL-17A inhibition through computer- from severe eosinophilic asthmatics (Supported by the Fondation de aided docking-based virtual screening. A18 specifically binds to a l’IUCPQ and the JD-Bégin Research Chair). region in the IL-17RA extracellular domain that overlaps with the Keywords: Asthma, Cough, Capsaicin, Allergen Challenge, TRPV1 binding site for IL-17A and this binding potently disrupts the IL-17A/ IL-17RA complex. We demonstrated that cyanidin effectively inhibited P21 IL-17A-mediated inflammatory gene expression in both human and Mechanisms of cross‑talk between pulmonary epithelial cells mouse cells. In the current study, we further investigated if A18 could and dendritic cells during type 2 inflammation attenuated airway inflammation in mouse models of steroid-resistant 1 1 1 1 Cecilia Forss , Peter Cook , Sheila Brown , Freya Svedberg , Katherine and severe asthma. 1 2 2 3 Stephenson , Margherita Bertuzzi , Elaine Bignell , Malin Enerbäck , Method: Adoptive transfer models for antigen‑induced airway 3 1 Danen Cunoosamy , Andrew M acdonald inflammation Manchester Collaborative Centre for Inflammation Research, Manches- OVA-specific Th17 and Th2 cells were transferred intravenously ter, United Kingdom; Manchester Fungal Infection Group, Manchester, (i.v.) into WT C57BL/6 female mice. Mice were challenged (i.n.) with United Kingdom; AstraZeneca R&D, Mölndal, Sweden OVA 1  day before transfer and 3 consecutive days after trans- 323–339 Correspondence: Cecilia Forss - cecilia.forss@postgrad.manchester.ac.uk fer. For A18 treatment, mice were injected (i.p.) with A18 on each Clinical and Translational Allergy 2017, 7(Suppl 2):P21 challenge day. Measurements were performed 24  h after the last challenge. Introduction: Everyday exposure to potentially harmful pathogens is High‑fat‑ diet (HFD) intervention carefully monitored and controlled by the interaction between a num- Starting at 4  weeks of age, WT C57BL/6 male mice were fed with ber of different cell types, such as epithelial cells and dendritic cells 10  kcal   % fat chow diet (CD) or a 60  kcal   % HFD for 14  weeks. A18 (DCs). Both have the ability to sense environmental changes, and initi- group of mice were injected (i.p.) with A18 daily for 4  weeks before ate and direct an immune response. This project is focusing on charac- subjection to measurements. terizing the interactions between epithelial cells and DCs in the early House dust mite (HDM)‑induced asthma stages of an allergen exposure. Aspergillus fumigatus (Af) is an oppor- WT C57BL/6 female mice were sensitized (s.c.) with HDM (100  µg/ tunistic fungus with the capacity to cause a range of responses, from mouse) in Complete Freund’s Adjuvant on day 0 and subsequently low-level inflammation in healthy people to life-threatening condi- challenged (i.n.) with HDM (100  µg/mouse) on day 14. A18 group of tions in the immunocompromised. mice were injected (i.p.) with A18 on days 13 and 14 and also adminis- Method: We have concentrated on understanding early events in the trated (i.n.) with A18 1 h before challenging. Measurements were per- immune response towards Af, with a current emphasis on DCs. formed 24 h after the last challenge. Results: 4 h post exposure, ~ 50% of in vitro generated DCs were posi- Results: tive for Af spores and the majority of the spores were internalised. Af A18 inhibited Th17-induced but not Th2-induced airway inflammation. positive DCs showed higher expression of CD40, CD80 and CD86 com- A18 substantially attenuated steroid-resistant obesity-associated pared to cells that were spore negative. Interestingly, while Af induced asthma. intermediate up-regulation of DC activation markers, it induced only A18 alleviated House dust mite (HDM)-induced neutrophilic airway low levels of cytokine production. As the type of interaction between inflammation. DCs and Af likely influences downstream activation and polarization of Conclusion: The findings strongly suggest that A18 (cyanidin) can be T cells, we are currently dissecting the impact of internalised vs. sur- used as the prototype for developing small molecule drugs for treat- face bound spores, compared to soluble fungal extracts, on activation ing the IL-17A-mediated severe asthma. of DCs in  vitro. In parallel, we have used murine GM-CSF- and Flt3-L- Keywords: Cyanidin, IL-17A, Severe Asthma derived DCs and human monocyte derived DCs, as well as human bronchial epithelial cells (HBECs) in air–liquid interface (ALI) cultures. P23 Additionally, we have used a murine model of intranasal exposure to Mechanisms involvement in CCL26 production by bronchial low doses of live Af to investigate Af uptake by, and activation of, pul- epithelial cells: importance in asthma and its severity monary DC subsets and epithelial cells in vivo. Marie-Chantal Larose, Anne-Sophie Archambault, Véronique Provost, Conclusion: Data generated so far indicate a fungal life stage-depend- Jamila Chakir, Michel Laviolette, Nicolas Flamand ent up-regulation of activation markers, accompanied by induction CRIUCPQ, Québec, Canada of low-level secretion of inflammatory cytokines. Af exposure in  vivo Correspondence: Marie-Chantal Larose - marie-chantal.larose@criucpq. resulted in marked airway inflammation with infiltration of immune ulaval.ca cells, goblet hyperplasia and increased collagen deposition. Ongoing Clinical and Translational Allergy 2017, 7(Suppl 2):P23 Clin Transl Allergy 2017, 7(Suppl 2):14 Page 13 of 17 Introduction: High pulmonary eosinophil counts correlate with Results: In this lung infection model, anti-Ym1 treatment led to a asthma severity and exacerbation. We recently showed that sputum reduction in IL-17A expression and a reduction in the number of CCL26 levels correlate with sputum eosinophils. We also found that innate  γδ T cells during the early stages of infection. The altered IL- among all CC chemokines, IL-13 selectively induced the expression of 17A response resulted in decreased IL-13 and IL-5 mRNA expression in CCL26 by bronchial epithelial cells (BECs) and this phenomenon is sig- whole lung tissue and reduced numbers of IL-5 + and IL-13 + CD4 + T nificantly enhanced in BECs from severe eosinophilic asthmatics. We cells and innate lymphoid cells; results mimicked by global IL-17A postulated that the superior CCL26 production that we observed in deficiency. severe asthmatics was the consequence of increased signaling events Conclusion: CLPs impaired the development of type 2 response by mediated by IL-13. We thus assessed the expression and functional modulating IL-17 expression. This suggests that CLPs can regulate responses of the different signaling effectors linked to the IL-13 signal- both innate and adaptive immune mechanisms that may be key in ing in CCL26 expression by BECs from healthy subjects, mild asthmat- tipping the balance of airway inflammation towards neutrophil domi- ics, and severe eosinophilic asthmatics. nance during allergic lung pathology. Method: Human primary BECs were isolated and cultured from bron- Keywords: Neutrophil, IL-17, Chitinase-Like Protein, Lung chial biopsies. BECs were treated with IL-13 or vehicle for different Inflammation times. Inhibitors or their vehicles were added to BECs 1 h before IL-13. References: CCL26 expression was assessed by qPCR and ELISA. STAT6 and phos- 1. Chupp GL et al. N Engl J Med. 2007;357:2016–27 phorylated (p)STAT6 were quantitated by ELISA in BEC lysates. SOCS3 2. Sutherland TE et al. Nat Immunol. 2014;15:1116–25 level were assessed by immunoblotting. Results: We confirmed the involvement of STAT6 in the induction of CCL26 expression by treating BECs with increasing STAT6 inhibitor. POSTER DISCUSSION SESSION 2—Treatment This led to a concentration-dependent inhibition of CCL26 expression in IL-13-stimulated BECs. In that regard, the pSTAT6/STAT6 ratios were P25 increased in IL-13-stimulated BECs from severe eosinophilic asthmat- Once‑daily tiotropium respimat add‑on therapy improves lung ics, compared to those from healthy subjects and mild asthmatics. This function in patients aged 6–17 years with severe symptomatic increased activation of STAT6 might be explained by decreased SOCS3 asthma level in BECs from severe eosinophilic asthmatics. Also, we confirmed 1 2 3 4 4 E. Hamelmann, C. Vogelberg , S. Goldstein , G. E. Azzi , M. Engel , R. the importance of STAT6 and IL-13 in CCL26 expression and produc- 5 6 Sigmund , S. J. Sz efler tion in IL-13-treated-BECs throughout the incubation time. 1 Children’s Center, Evangelisches Krankenhaus Bielefeld, and Allergy Keywords: CCL26, Eosinophils, Bronchial Epithelial Cells 2 Center of the Ruhr University, Bochum, Germany; University Hospital Carl Gustav Carus, Technical University of Dresden, Dresden, Germany; Island P24 Medical Research, Rockville Centre, New York, New York City, New York, Chitinase‑like proteins: the missing link in allergen‑induced United States; Boehringer Ingelheim Pharma GmbH & Co. KG, Ingelheim neutrophilic inflammation? Am Rhein, Germany; Boehringer Ingelheim Pharma GmbH & Co. KG, 1 2 2 3 6 Nicola Logan , Dominik Ruckerl , Judith E. Allen , Tara E. Sutherland Biberach An Der Riss, Germany; Children’s Hospital of Colorado and the Institute of Immunology and Infection Research, University of Edin- University of Colorado School of Medicine, Aurora city, Colorado, United burgh, Edinburgh, United Kingdom; Faculty of Biology, Medicine States & Health, University of Manchester, Manchester, United Kingdom; Correspondence: Eckard Hamelmann - eckard.hamelmann@evkb.de Manchester Collaborative Centre for Inflammation Research, University Clinical and Translational Allergy 2017, 7(Suppl 2):P25 of Manchester, Manchester, United Kingdom Correspondence: Tara E. Sutherland - tara.sutherland@manchester.ac.uk Introduction: Tiotropium Respimat (tioR) add-on therapy to inhaled Clinical and Translational Allergy 2017, 7(Suppl 2):P24 corticosteroids (ICS) with or without additional controllers has been shown to improve lung function in Phase II and III studies of adults, adolescents and children with symptomatic asthma. We present a Introduction: Immune responses during asthma are typically consid- pooled analysis of lung function data in adolescents and children with ered as T-helper 2 (Th2) type responses marked by increased eosino- severe symptomatic asthma. phils. However, inflammation in severe asthmatics is often defined by Method: Two Phase III, randomised, double-blind, placebo-controlled, neutrophil or mixed neutrophil/eosinophil airway inflammation, along parallel-group, 12-week trials in patients aged 6–11  years (Viva- with increased IL-17 cytokine levels and airway remodelling. Little is TinA-asthma; NCT01634152) and 12–17  years (PensieTinA-asthma; known about the triggers that make neutrophils/IL-17 dominant char- NCT01277523) with severe symptomatic asthma. Patients received acteristics in severe asthmatics, or how these responses contribute to once-daily tioR 5 µg (2 × 2.5 µg), tioR 2.5 µg (2 × 1.25 µg) or placebo pathology, especially in scenarios when Th2-inflammation remains Respimat (pboR) as add-on to high-dose ICS plus another control- prevalent. Chitinase-like proteins (CLPs) are established markers of ler or as add-on to medium-dose ICS plus two other controllers. ICS immune activation and pathology. Of note, whilst considered Th2- dose was as defined in GINA 2009 (PensieTinA) or 2010 (VivaTinA) driven molecules, CLPs are also associated with IL-17 and neutrophilic guidelines. Patients were required to have a  ≥  3-month (PensieTinA) inflammation. In asthmatic patients, enhanced levels of circulating or  ≥  6-month (VivaTinA) history of asthma and be symptomatic at CLP YKL-40 correlates with disease severity and airway neutrophilia . screening and before randomisation by Asthma Control Questionnaire In addition, we recently described a novel role for murine CLPs in (interviewer-administered; VivaTinA) mean score  ≥  1.5. Primary end driving alveolar neutrophil recruitment through increased IL-1/IL-18 point of both studies: change from baseline (response) in peak forced expression and expansion of IL-17A-producing γδ T cells . Consider- expiratory volume in 1  s within 3  h post-dose (peak FEV ); key 1(0–3h) ing their association with both Th2 pathology and IL-17A/neutrophils, secondary end point: trough FEV response (measured 10 min before we sought to examine whether CLPs might link IL-17 responses to Th2 next dose of study medication); post hoc end point: trough FEV / inflammation. forced vital capacity (FVC) ratio, all measured at Week 12. Method: We explored the effects of neutralising the most abun- Results: 793 participants (VivaTinA: 401; PensieTinA: 392) were ran- dant murine CLP Ym1, with monoclonal antibody treatment during domised across both trials; 792 were included in this pooled full infection with the rodent lung migrating nematode N. brasiliensis. analysis set. Baseline demographics and disease characteristics were Infection with N. brasiliensis  in mice elicits lung immune responses balanced between treatment groups. TioR add-on therapy improved similar to that observed in allergic airway inflammation, thereby mak - lung function in the pooled population at Week 12, with tioR 5  µg ing it a valid model to study the interaction between IL-17A and Th2 showing superior improvements in peak FE V response, trough 1(0–3h) inflammation. Clin Transl Allergy 2017, 7(Suppl 2):14 Page 14 of 17 Table 3 Treatment results Response (change Peak Trough FEV Trough FEV / 1 1 from baseline) FEV response (mL) FVC ratio (%) 1(0–3h) measured response at Week 12 (mL) Tiotropium Respimat 117 ± 34 71 ± 34 1.921 ± 0.666 5 µg once daily (51, 183) (3, 139) (0.614, 3.229) (n = 260), adjusted p = 0.0005 p = 0.0395 p = 0.0040 mean of difference versus placebo Respimat ± SE Figure 6 Patient 1 and patient 2 (95% CI) Tiotropium Respimat 74 ± 33 64 ± 34 1.930 ± 0.666 2.5 µg once daily (8, 140) (−3, 132) (0.623, 3.236) (n = 263), adjusted p = 0.0273 p = 0.0617 p = 0.0038 ventilation, IMV was initiated. On Day-1, despite medical therapy and mean of difference IMV optimizations, severe dynamic hyperinflation with refractory versus placebo hypoxemia persisted and VV-ECMO was initiated. Thoracic CT revealed Respimat ± SE almost complete bilateral lung collapse. Methicillin-sensitive S. aureus (95% CI) and S. pneumoniae were isolated in endotracheal aspirate. Normal Full analysis set. CI, confidence interval; SE, standard error lung function was progressively reestablished following considerable viscid mucus secretions elimination and antibiotherapy. ECMO-VV was discontinued on Day-18. Extubation, ICU and hospital discharges occurred on Day-24, -27 and -55, respectively. FEV response and FEV /FVC ratio versus pboR, and tioR 2.5 µg show- 1 1 Conclusion: How this report contributes to current knowledge. Dur- ing superior improvements in peak FE V response and FEV / 1(0–3h) 1 ing refractory NFA, low-flow ECCO R allows correction of severe res- FVC ratio versus pboR, with numerical improvements in trough FEV piratory acidosis and facilitates extubation while high-flow VV-ECMO response versus pboR (Table). Safety and tolerability of tioR in both tri- is needed when significant atelectasis complicates airway narrowing als was comparable to placebo. and severe hypoxemia ensues. Knowledge of respiratory failure patho- Conclusion: Tiotropium Respimat add-on therapy is an effective physiology in refractory NFA allows the correct use of different ECLS bronchodilator, producing clinically meaningful improvements ver- modalities as a bridge to recovery. sus placebo in lung function in patients aged 6–17  years with severe symptomatic asthma, mirroring findings in adult patients with symp - tomatic asthma. Consent to publish: Consent to publish was received from the Keywords: Asthma Management, Bronchodilators, Children patients. P26 P27 Near‑fatal asthma: differential role of ECCO2R and VV‑ECMO Clinical and functional characteristis of 4 severe asthmatic as rescue therapies patients with absence of response to omalizumab 1 2 2 2 Raquel Mesquita , Luis Coentrão, Rui Veiga , José-Artur Paiva , Roberto Wendy Vargas Porras, Ana González Moreno, Jesus Macías Iglesias, Roncon-Albuquerque Jr Gustavo Córdova Ramos, Yesenia Peña Acevedo, Miguel Angel Tejedor Internal Medicine Department, Centro Hospitalar de São João, Porto, Alonso, Maria Del Mar Moro Moro Portugal; Emergency and Intensive Care Medicine Department, Centro University Hospital Foundation Alcorcón, Madrid, Spain Hospitalar de São João, Porto, Portugal Correspondence: Wendy Vargas Porras - wendy.drawenwen@gmail.com Correspondence: Raquel Mesquita - raquelmsmesquita@gmail.com Clinical and Translational Allergy 2017, 7(Suppl 2):P27 Clinical and Translational Allergy 2017, 7(Suppl 2):P26 Introduction: 69.9% of patients with severe asthma respond to Introduction: During a near-fatal attack of asthma (NFA) alveolar omalizumab, when it is used according to International Guidelines of hypoventilation underlies respiratory acidosis while severe hypoxemia asthma. indicates intrapulmonary shunt. Illustrate the differential role of low- We describe clinical and functional characteristics of 4 patients with flow extracorporeal CO removal (EC CO R) and high-flow veno-venous severe asthma treated with omalizumab and belong to our series of 2 2 extracorporeal membrane oxygenation (VV-ECMO) as rescue therapies severe asthmatic patients treated with this drug. in NFA. Method: We collected 24 patients with severe asthma treated with Method: Patient 1 A 31-year-old male with early-onset asthma and omalizumab from the Allergy Unit of the University Hospital Founda- epilepsy was admitted with acute respiratory failure (ARF) compli- tion Alcorcón, during the period 2007 to 2015. Of these, 4 patients did cated by generalized tonic–clonic seizure. He presented severe acute not respond to treatment. The “No responders”  was defined by the respiratory acidosis without hypoxemia after intubation (pH-7.10 responsible doctor, based on the clinical and functional assessment of pCO -82  mmHg P/F 556). Thoracic CT: diffuse bronchial wall thick - each patient considering several clinical and functional characteristics: ness without atelectasis. On Day-5 dynamic hyperinflation (auto- beta-adrenergic use, number of exacerbations and scores ACQ and PEEP 10 cmH O) under deep sedation and neuromuscular blockade ACT, other comorbidities, spirometric and lung volumes. These values persisted, precluding weaning from invasive mechanical ventilation were measured in the last year before the start of omalizumab and the (IMV). ECCO R was initiated. Extubation, active physical therapy and final year of treatment with omalizumab. full patient mobilization were possible on Day-6. EC CO R discon- Results: In 4 patients it was withdrawn omalizumab because of no tinuation, ICU and hospital discharges occurred on Day-8, -9 and -11, efficacy. Responders and non-responders were treated, at least, during respectively. 1  year. The non-responders had more non-respiratory comorbidities Results: Patient 2 A 34-year-old obese female with late-onset and than responders (75%-25%,p  =  0.07). However responders, seemed poorly controlled asthma was admitted with ARF with severe hypox- to have more severe asthma, with a greater number of exacerbations emia (pH-7.45 pCO -30  mmHg P/F 69). After a trial of non-invasive (responders 3.24 vs no responders 2.75), more use of beta-adrenergic 2 Clin Transl Allergy 2017, 7(Suppl 2):14 Page 15 of 17 (responders 2.47 vs no responders 1), more cycles oral corticoster- However, determination of inflammation using patient-based symp - oids (responders 2.64 vs no responders 1.5) and higher ACQ score tom assessment or spirometry is difficult and frequently underesti- (responders 2.1 vs no responders 1.8). Only use of oral corticosteroids mates asthma severity. Measurement of exhaled nitric oxide (FeNO) and use of beta-adrenergic the differences were significant (p  =  0.02 can support the clinical assessment of the patient at the point of and 0.05 respectively). However the residual volume  (RV) higher in care and can provide insights into underlying airway inflammation non-responders (responders 188% vs no responders 154%), although and therefore potentially help practitioners optimize their treatment differences were not statistically significant (p = 0.3). decisions. High FeNO concentrations have been associated with more In the last year of use of omalizumab, asthma was worse in the group severe/uncontrolled asthma and poor compliance with ICS therapy. of non-responders, with increased use of beta-adrenergic (p =  0.005) Method: The objective of the ASTH MA survey was to explore the real and greater use of steroids cycles (p = 0.002). world impact of measuring FeNO on physicians’ treatment decisions. Conclusion: In non-responders patients, the RV was higher and also Physicians initially recorded their assessment of airway inflammation had more comorbidities. (low, intermediate or high) based on the patient’s clinical presentation The small number of our sample limits the validity of our findings. and then by measuring patient’s FeNO using an approved electro- chemical device. Based on the FeNO result, physicians recorded what changes in drug therapy were made. P28 Results: Data from 337 physician practices, which included 7,901 One year follow‑up of patients after omalizumab treatment patients with asthma, were available for analysis. Assessment of withdrawal inflammation using clinical impression vs FeNO measurement was Irena Krcmova, Jakub Novosad determined to be low in 4,247 patients (53.8%) vs 5,083 (64.3%) Institute of Clinical Immunology and Allergy, University Hospital, Hradec (< 25 ppb); intermediate in 2,749 (34.8%) vs 1,802 (22.8%) (25–50 ppb) Kralove, Czech Republic and high in 905 (11.5%) vs 1016 (12.9%) (> 50 ppb). Mean high FeNO Correspondence: Irena Krcmova - irena.krcmova@fnhk.cz was 87  ppb (range 51–300, median 77) with FeNO  ≥  51–99  ppb in Clinical and Translational Allergy 2017, 7(Suppl 2):P28 n  =  760, FeNO 100–199  ppb in n  =  229 and FeNO 200–300  ppb in n = 27 patients. Clinical impression matched actual FeNO of < 25 ppb Introduction: Omalizumab has demonstrated its clinical efficacy in in 64.4%, 25–50  ppb in 46.9% and  >  50  ppb in only 33.6%. Changes asthmatic patients in number of studies. However, data about its clini- in treatment were made in 68.4% (695/1016) of patients who had a cal benefits after treatment cessation are still missing. To stress this high FeNO (>  50  ppb); stepping up anti-inflammatory treatment in challenging clinical issue, we have arranged a pilot observational- 96.1% and stepping down in 2.9%. Inhaled steroids were started in 345 study following clinical and laboratory data of 12 patients with severe patients and increased in 213 patients. Oral steroids were increased/ uncontrolled atopic asthma treated with omalizumab (11–61 months) started in 110 patients. Omalizumab was started in 2 patients. for one year after treatment withdrawal due to clinical stability. Conclusion: Our results suggest that measurement of FeNO at point Method: Study had a mixed design with repeated measures at 4 time of care improves the clinical assessment of underlying airway inflam- points, at treatment initiation, directly after treatment cessation, and mation in asthma and leads to clinically relevant changes in treatment 6 and 12  months thereafter. We have focused on clinical (pulmonary especially in those patients presenting with high FeNO (> 50 ppb). functions, inhalation corticosteroid (ICS) doses, asthma control test Keywords: FeNO, Asthma, Management, ASTH2MA Survey (ACT ), skin prick tests positivity) and laboratory (FeNO, eosinophils and total IgE levels) parameters. Statistical analysis of collected data has P30 been undertaken. Repeated measures were treated using a general 4‑month omalizumab efficacy outcomes for inadequate linear model or Friedman ANOVA in case of normality assumption vio- controlled severe allergic asthma in the Netherlands 2012–2015 lation. Post-hoc analysis was applied using Wilcoxon signed-rank test Sanne Snelder, Gert-Jan Braunstahl with Bonferroni adjustment. Sint Franciscus Gasthuis, Rotterdam, The Netherlands Results: We have demonstrated a significant reduction of ICS doses Correspondence: Sanne Snelder - sannesnelder@hotmail.com and SPT positivity (wheal diameter/mm) and increase of ACT dur- Clinical and Translational Allergy 2017, 7(Suppl 2):P30 ing omalizumab treatment regardless of a dominant allergic linkage, treatment duration or omalizumab dose. This effect has been visible even 6  months after treatment withdrawal, but only ACT sustained Introduction: Since 2006, omalizumab has been prescribed for inad- increased significantly 12  months after treatment cessation. Two equately controlled severe allergic asthma in the Netherlands. In 2011, patients had a treatment of oral corticosteroids, which were perma- more stringent rules were applied by Dutch healthcare policy makers. nently tapered during the omalizumab therapy and subsequently. This implied that omalizumab should be strictly prescribed within the Conclusion: We conclude, that omalizumab treatment outcomes registered label in exchange for reimbursement: inclusion criteria were reach clinical and statistical significance even one year after treatment a positive skin test or in  vitro activity to a perennial aeroallergen, a withdrawal. “Step down” of omalizumab in “treatment responders” FEV1 less than 80 percent, more than 2 severe exacerbations and sub- remains a highly individual process that can undergo by mutual agree- stantial symptoms despite treatment with inhaled corticosteroids (ICS) ment with the patient under clearly defined treatment plan. and long-acting B2-agonists (LABAs). Keywords: Asthma, Omalizumab, Withdrawal, Clinical Effect To evaluate the 4-month efficacy outcomes of omalizumab treatment for inadequate controlled severe allergic asthma after the stringent rules were applied by the Dutch healthcare policy. P29 Method: This is a “real world” prospectively designed data registry High FeNO leads to changes in asthma treatment: cohort analysis in which the outcomes of patients (2012–2015) who received omali- from the assessment of TH2 inflammation (ASTH2MA) survey 1 2 2 2 zumab were evaluated after the policy change. The primary endpoint Nicola Alexander Hanania , Marc Massanari , Heike Hecker , Eric Kassel , 3 4 was a good or excellent response to omalizumab after 16  weeks as Craig Laforce , Kathy Rickard 1 2 scored by the treating physician. Secondary endpoints include change Baylor College of Medicine, Houston, Texas, United States; Circassia, 3 from baseline to week 16 in the Asthma Control Questionnaire (ACQ), Raleigh, North Carolina, United States; North Carolina Clinical Research, 4 FEV1 scores and oral corticosteroids (OCS) use. Raleigh, North Carolina, United States; Circassia, Raleigh, North Carolina, Results: 403 patients had a complete data set and could be evalu- United States ated. 63% percent of the patients had a good or excellent response Correspondence: Heike Hecker - heike.hecker@circassia.com to omalizumab after 16 weeks. 85.5% of the responders showed more Clinical and Translational Allergy 2017, 7(Suppl 2):P29 than 0.5 points improvement in the ACQ score at 16 weeks. The mean FEV increased from 71.58 to 79.06. 63% of the responders had an Introduction: Recognizing TH inflammation in asthma has led to improvement of 5% of the FEV1. The maintenance OCS use was lower improvements in optimization of drug therapy and disease control. at 16  weeks. There was no relation between FEV1  <  80 and ≥  80% at Clin Transl Allergy 2017, 7(Suppl 2):14 Page 16 of 17 Table 4 Systematic review Trial Kuehr Casale Kopp 2008 Massanari et al 2006 2010 Disease Seasonal Seasonal Seasonal rhinitis persistent rhinitis rhinitis and asthma asthma Severity Moder- Moderate mild asthma At least ate- moderate severe Age range 6–17 18–50 11–46 18–55 Total 221 159 140 275 number of patients Site of study Germany USA Germany USA Duration of 36 weeks 22 weeks 20 weeks 26 weeks enrolment Figure 7 Improvement of ACQ and FEV1 compared to response Allergen Grass and aqueous Depigoid,modified Cat, dog used birch short grass pollen and HDM extract rag- extract extract Table 3 Response vs FEV1 < 80 weed extract Response Protocol of Conven- Rush Pre-seasonal rush Cluster immuno- tional protocol titration protocol immuno- YES NO Total therapy therapy FEV1 < 80 YES 178 80 258 Length of 14 weeks 10 weeks 8 weeks Perennial pre- allergen FEV1 < 80 NO 73 33 106 seasonal Total 251 113 364 immuno- P = 0.981 therapy Time of 12 week Pre-treat- Co-administration Pre-treat- adminis- after ment ment for Table 4 Response vs year of inclusion tration of immu- 9 weeks 13 weeks Omali- nother- Year of inclusion Response No response zumab apy started 2012 (%) 63.6 36.4 Dose of 0.016 mg/ 0.016 mg/ Dose/kg/IgE IU/mL 0.016 mg/ 2013 (%) 70.7 29.3 Omali- kg per kg/IgE every 2–4 weeks kg/IgE 2014 (%) 64.9 35.1 zumab IU/Ml IU/mL IU/mL every 2 or 2015 (%) 69.2 30.8 4 weeks P = 0.690 Introduction: Oral corticosteroids (OCS) are used in the management baseline and response after 4  months (Table  3). The response rate of both acute and severe asthma. Acute courses are typically well toler- remained stable over the years 2012–2015 (Table  4). Figure  1 shows ated, but well documented side effects occur in prolonged treatment. that most of the responders had an improvement of both FEV1 and Side effects include skin thinning, cataracts, diabetes, osteopenia and ACQ. It also shows that ACQ appears to be a better measurement for a easy bruising; these may impact adherence to treatment. Patients response than FEV1. rarely admit poor adherence to treatment but, with increasing avail- Conclusion: 63% of the 403 patients with inadequately controlled ability of monoclonal antibody therapy, adherence should be good severe allergic asthma had a good or excellent response to omali- prior to escalation to high-cost treatment. Mass-spectrometry can zumab after 16  weeks. Overall the ACQ improved, FEV1 increased assess concentrations of cortisol and prednisolone in peripheral blood, and there was lower use of OCS at 16  weeks. There was no relation- and this can provide evidence of OCS adherence as well as assessing ship between patients with a FEV1 < 80 and ≥ 80% at baseline and the absorption. We conducted an audit of results of all our prednisolone response rate. Improvement of ACQ appears to be a better measure- assays conducted in patients with severe asthma. ment for response than improvement of FEV1. Method: Data was collected on all patients with severe asthma under- Keywords: Asthma, Omalizumab going prednisolone assays at our centre. OCS assays were performed prior to escalation of therapy where there was a suspicion of poor P31 adherence. The results of the assay, participant age, gender, BMI, FEV1, Adherence to oral corticosteroids; audit results of prednisolone Fractional Exhaled Nitric Oxide (FeNO), eosinophil count and predniso- assays lone dose were recorded. These were used to determine which patient T. L. Jones, D. Neville, E. R. Heiden, E. Lanning, T. Brown, H. Rupani, K. S. characteristics correlate with adherence. Babu, A. J. Chauhan Results: 14 adults taking oral prednisolone were included in this Portsmouth Hospitals NHS Trust, Portsmouth, United Kingdom audit. This comprised 11 females and 3 males with a mean age of Correspondence: Thomas Llewelyn Jones - thomas.jones@porthosp.nhs.uk 46.3  ±  13.4  years, a mean prednisolone dose of 20.2  ±  10.4  mg, a Clinical and Translational Allergy 2017, 7(Suppl 2):P31 Clin Transl Allergy 2017, 7(Suppl 2):14 Page 17 of 17 mean BMI of 30.7  ±  7.2, a mean FeNO of 79.9  ±  53.2, mean FEV1 of The purpose of this review is to establish whether the combination of 64 ±  17.6% predicted and a mean eosinophil count of 0.35 ±  0.34. 5 omalizumab and subcutaneous allergen immunotherapy (SCIT ) has an out of 14 were compliant as assessed by a supressed cortisol (5/5) or add-on benefit to the efficacy and safety of SCIT in asthma and allergic detectable blood prednisolone prior to dosing (4/5). All 14 patients rhinitis? absorbed prednisolone well following oral OCS administration. Factors Method: This systematic review included double-blind randomized correlating with compliance included gender (males 66% compliant, placebo controlled trials of the add-on benefits of the combination of females 27% compliant), BMI (25.5 kg/m2 compliant vs 33.5, p = 0.05) omalizumab and SCIT compared to SCIT alone. and peripheral eosinophil count (compliant 0.15 vs 0.45, p = 0.09). Results: The combination of omalizumab and SCIT has decreased Conclusion: Only 36% of patients in our audit were adherent to OCS symptoms score, rescue medication score, and symptoms load in highlighting the importance of checking medication adherence dur- patients with rhinitis and patients with rhinitis and co-existing asthma. ing each clinical review, particularly when considering patients for In patients with asthma and those with rhinitis treated with rush novel therapies. Male gender, lower BMI and low eosinophil count immunotherapy, the percentage of patients developed systemic reac- were associated with compliance, but FeNO, FEV1 and OCS dose were tion to SCIIT including severe reaction requiring adrenaline admin- not. We suggest an objective assessment of compliance prior to initia- istration was significantly decreased in the combination treatment tion of expensive novel biological agents. group compared with SCIT/placebo group. Omalizumab has reduced Keywords: Prednisolone, Medication, Adherence, Education the risk for anaphylaxis by 5 folds in rhinitis patients treated with rush immunotherapy protocol. Only one trial reported the of number of adrenaline doses used during the trial P32 Conclusion: The addition of omalizumab has increased the efficacy of Does the combination of omalizumab and Subcutaneous allergen SCIT in patients with rhinitis and patients with rhinitis and co-existing immunotherapy (SCIT) have an add‑on benefit to the efficacy asthma. Omalizumab also reduced the risk of serious systemic reaction and safety of allergen immunotherapy in asthma and allergic in patients with asthma and in rhinitis patients treated with rapid high rhinitis? systematic review 1 2 3 2 dose of allergen. Addition of omalizumab has helped more patients at M. Y. Eldegeir , A. A. Chapman , M. Ferwana , M. Caldron 1 2 high risk of adverse effects related to SCIT to reach their target immu- National Guard Hospital, Dammam, Saudi Arabia; Imperial Collage 3 notherapy. An important findings of this review is that the use of London, London, United Kingdom; King Saud University, Riyadh, Saudi adrenaline is under-reported by immunotherapy clinical trials Arabia Keywords: Omalizumab, Immunotherapy, Asthma, Rhinitis, Allergic Correspondence: Manal Yousif Eldegeir - almanjil@gmail.com Clinical and Translational Allergy 2017, 7(Suppl 2):P32 Publisher’s Note Introduction: The last two decades witnessed a substantial expansion Springer Nature remains neutral with regard to jurisdictional claims in pub- in our understanding of the pathoimmunological mechanism of aller- lished maps and institutional affiliations. gic respiratory diseases. This led to a huge surge of interest in devel- oping novel treatment modalities aiming for improved efficacy and safety of AIT. Co-administration of anti IgE is an attractive approach with the added benefit of omalizumab as a biological agent which has its own independent Immunomodulation effect on asthma and rhinitis. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Clinical and Translational Allergy Springer Journals

Abstracts from the 3rd International Severe Asthma Forum (ISAF)

Ketelaar, M.; De Kant, K.; Dijk, F.; Klaassen, E.; Grotenboer, N.; Nawijn, M.; Dompeling, E.; Koppelman, G.; Murray, Clare; Foden, Philip; Lowe, Lesley; Durrington, Hannah; Custovic, Adnan; Simpson, Angela; Simpson, Andrew; Shaw, Dominick; Sousa, Ana; Fleming, Louise; Roberts, Graham; Pandis, Ioannis; Bansal, Aruna; Corfield, Julie; Wagers, Scott; Djukanovic, Ratko; Chung, Kian; Sterk, Peter; Vestbo, Jorgen; Fowler, Stephen; Tebbutt, S.; Singh, A.; Shannon, C.; Kim, Y.; Yang, C.; Gauvreau, G.; Fitzgerald, J.; Boulet, L.; O’Byrne, P.; Begley, N.; Loudon, A.; Ray, D.; Baos, Selene; Cremades, Lucía; Calzada, David; Lahoz, Carlos; Cárdaba, Blanca; Asosingh, Kewal; Lauruschkat, Chris; Queisser, Kimberly; Wanner, Nicholas; Weiss, Kelly; Xu, Weiling; Erzurum, Serpil; Sokolowska, Milena; Chen, Li-Yuan; Liu, Yueqin; Martinez-Anton, Asuncion; Logun, Carolea; Alsaaty, Sara; Cuento, Rosemarie; Cai, Rongman; Sun, Junfeng; Quehenberger, Oswald; Armando, Aaron; Dennis, Edward; Levine, Stewart; Shelhamer, James; Choi, Kilyong; Lazova, Snezhina; Perenovska, Penka; Miteva, Dimitrinka; Priftis, Stamatios; Petrova, Guergana; Yablanski, Vassil; Vlaev, Evgeni; Rafailova, Hristina; Kumae, Takashi; Holmes, L.; Yorke, J.; Ryan, D.; Chinratanapisit, Sasawan; Matchimmadamrong, Khlongtip; Deerojanawong, Jitladda; Karoonboonyanan, Wissaroot; Sritipsukho, Paskorn; Youroukova, Vania; Dimitrova, Denitsa; Slavova, Yanina; Lesichkova, Spaska; Tzocheva, Iren; Parina, Snezhana; Angelova, Svetla; Korsun, Neli; Craiu, Mihai; Stan, Iustina; Deliu, Matea; Yavuz, Tolga; Sperrin, Matthew; Sahiner, Umit; Belgrave, Danielle; Sackesen, Cansin; Kalayci, Ömer; Velikov, Petar; Velikova, Tsvetelina; Ivanova-Todorova, Ekaterina; Tumangelova-Yuzeir, Kalina; Kyurkchiev, Dobroslav; Megremis, Spyridon; Constantinides, Bede; Sotiropoulos, Alexandros; Xepapadaki, Paraskevi; Robertson, David; Papadopoulos, Nikolaos; Wilkinson, Maxim; Portsmouth, Craig; Ray, David; Goodacre, Royston; Valerieva, Anna; Bobolea, Irina; Vera, Daiana; Gonzalez-Salazar, Gabriel; Moreno, Carlos; Rodriguez, Consuelo; Las Cuevas Moreno, Natividad; Wang, R.; Satia, I.; Niven, R.; Smith, J.; Southworth, T.; Plumb, J.; Gupta, V.; Pearson, J.; Ramis, I.; Lehner, M.; Miralpeix, M.; Singh, D.; Satia, Imran; Woodhead, Mark; O’Byrne, Paul; Smith, Jaclyn; Forss, Cecilia; Cook, Peter; Brown, Sheila; Svedberg, Freya; Stephenson, Katherine; Bertuzzi, Margherita; Bignell, Elaine; Enerbäck, Malin; Cunoosamy, Danen; Macdonald, Andrew; Liu, Caini; Zhu, Liang; Fukuda, Kiochi; Zhang, Cunjin; Ouyang, Suidong; Chen, Xing; Qin, Luke; Rachakonda, Suguna; Aronica, Mark; Qin, Jun; Li, Xiaoxia; Larose, Marie-Chantal; Archambault, Anne-Sophie; Provost, Véronique; Chakir, Jamila; Laviolette, Michel; Flamand, Nicolas; Logan, Nicola; Ruckerl, Dominik; Allen, Judith; Sutherland, Tara; Hamelmann, E.; Vogelberg, C.; Goldstein, S.; Azzi, G.; Engel, M.; Sigmund, R.; Szefler, S.; Mesquita, Raquel; Coentrão, Luis; Veiga, Rui; Paiva, José-Artur; Roncon-Albuquerque, Roberto; Porras, Wendy; Moreno, Ana; Iglesias, Jesus; Ramos, Gustavo; Acevedo, Yesenia; Alonso, Miguel; Mar Moro Moro, Maria; Krcmova, Irena; Novosad, Jakub; Hanania, Nicola; Massanari, Marc; Hecker, Heike; Kassel, Eric; Laforce, Craig; Rickard, Kathy; Snelder, Sanne; Braunstahl, Gert-Jan; Jones, T.; Neville, D.; Heiden, E.; Lanning, E.; Brown, T.; Rupani, H.; Babu, K.; Chauhan, A.; Eldegeir, M.; Chapman, A.; Ferwana, M.; Caldron, M.
Clinical and Translational Allergy , Volume 7 (2) – May 24, 2017

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Springer Journals
Copyright
Copyright © 2017 by The Author(s)
Subject
Medicine & Public Health; Allergology; Immunology; Pneumology/Respiratory System
eISSN
2045-7022
DOI
10.1186/s13601-017-0149-8
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Clin Transl Allergy 2017, 7(Suppl 2):14 Clinical and DOI 10.1186/s13601-017-0149-8 Translational Allergy Open Access MEE TING ABSTR AC TS Abstracts from the 3rd International Severe Asthma Forum (ISAF) Manchester, United Kingdom. 17 November 2016–19 November 2016 Published: 24 May 2017 ORAL ABSTRACT SESSION 1—Asthma: from mechanisms (IL-1RL1-a alone: AUC = 0.65 [95 CI 0.52–0.79, P = 0.04], API + IL-1RL1- to management a: AUC = 0.70 [95 CI 0.56–0.84, P = 0.01]). Interestingly, IL-1RL1-a levels had a negative direction of effect. Conclusion: Our study shows that serum IL-1RL1–a levels measured in O01 wheezing children at age 2–3 years do not predict doctors’ diagnosed Serum IL‑1RL1‑A levels predict an eosinophilic subtype of asthma asthma as general phenotype at age 6 years, but negatively predict an in preschool wheezing children 1 2 1 3 eosinophilic subphenotype of asthma. M. E. Ketelaar , K. Van De Kant , F. N. Dijk , E. M. M. Klaassen , N. 4 4 2 1 This suggests that IL-1RL1 might play a protective role in the devel- Grotenboer , M. C. Nawijn , E. Dompeling , G. H. Koppelman 1 opment of eosinophilia in children who experience asthma at school University Medical Center Groningen, Beatrix Children’s Hospital, Gron- age and implies that IL-1RL1 targeted therapy could rather be further ingen Research Institute for Asthma and COPD (GRIAC), Groningen, The 2 explored in the subphenotype of asthmatic children with predomi- Netherlands; Department of Pediatric Pulmonology, School for Public nant eosinophilic inflammation. Health and Primary Care (CAPHRI), Maastricht University Medical Center 3 Keywords: Childhood Asthma, Eosinophilic Asthma, Prediction, (MUMC+), Maastricht, The Netherlands; Department of General Practice, IL-1RL1, Serum School for Public Health and Primary Care (CAPHRI), Maastricht University Medical Center (MUMC+), Maastricht, The Netherlands; University Medi- cal Center Groningen, Department of Pathology and Medical Biology, Laboratory of Experimental Pulmonology and Inflammation Research (EXPIRE), Groningen Research Institute for Asthma and COPD (GRIAC), Groningen, The Netherlands Correspondence: Maria Elizabeth Ketelaar - m.e.ketelaar@student.rug.nl Clinical and Translational Allergy 2017, 7(Suppl 2):O01 Introduction: Respiratory symptoms are common in preschool chil- dren. However, which of these wheezers will develop asthma at school age, and what phenotype they will develop remains difficult to pre - dict. Current models such as the asthma prediction index (API) are based on clinical parameters and have only modest predictive accu- racy. Expression levels of well replicated asthma genes could poten- tially form novel biomarkers for asthma prediction. IL1RL1 is an asthma susceptibility gene, and has also been linked to eosinophilia. Therefore, we hypothesized that expression levels of IL1RL1 in the form of soluble IL-1RL1-a measured in serum from wheezing preschool chil- dren contribute to the prediction of asthma at school age. Moreover, since IL1RL1 was previously associated with blood eosinophilia, our second aim was to determine whether serum IL-1RL1-a levels predict eosinophilic asthma. Method: We used logistic predictive modeling in a prospective Dutch Figure 1 Prediction of eosinophilic childhood using IL-1 RL1-a serum birth cohort (n  =  202 wheezers), and calculated the area under the levels and the API curve (AUC) of the sensitivity/1-specificity curves of potential models. Results: Neither IL-1RL1-a serum levels at age 2–3  years alone nor its combination with the API had predictive value for doctors’ diag- O03 nosed asthma at age 6y (IL-1RL1-a alone: AUC = 0.50 [95 CI 0.41–0.59, Diagnosing asthma in symptomatic children using lung function: P = 0.98], API + IL-1RL1-a: AUC = 0.57 [95 CI 0.49–0.66, P = 0.12]). evidence from a birth cohort study However, IL-1RL1-a serum levels at age 2–3  years correlated with the 1 1 1 1 Clare Murray , Philip Foden , Lesley Lowe , Hannah Durrington , severity of airway eosinophilia (determined by levels of exhaled frac- 2 1 Adnan Custovic , Angela Simpson tion of NO, [FeNO]) in children who had developed asthma at age 6y 1 2 University of Manchester, Manchester, United Kingdom; Imperial (Pearson’s R  =  −0.24, P  =  0.046, N  =  59). Logistic predictive mod- College, London, United Kingdom eling of eosinophilic asthma at age 6y (asthma with FeNO ≥  20  ppb) Correspondence: Angela Simpson - angela.simpson@manchester.ac.uk showed that IL-1RL1-a serum levels itself and in combination with Clinical and Translational Allergy 2017, 7(Suppl 2):O03 the API could predict this eosinophilic subphenotype of asthma © The Author(s) 2017. This article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. 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. Clin Transl Allergy 2017, 7(Suppl 2):14 Page 2 of 17 Introduction: In the UK, new national draft guidance for the diagno- Hospital of South Manchester, NHS Foundation Trust, Manchester, United sis of childhood asthma proposes algorithms based on four tests of Kingdom; Respiratory Research Unit, University of Nottingham, Notting- lung function, each used as a dichotomous variable (FE V /FVC ratio ham, United Kingdom; Respiratory Therapeutic Unit, GSK, Stockley Park, less than the lower limit of normal [LLN], bronchodilator reversibility London, United Kingdom; National Heart and Lung Institute, Imperial [BDR] ≥ 12%, FeNO ≥ 35 ppb and PEFR variability). However, accuracy College, London, United Kingdom; NIHR Southampton RespiratoryBio- of these tests in diagnosing asthma in children is unknown, as the medical Research Unit, Clinical and Experimental Sciences and Human evidence is largely derived from studies of adults. Within the setting Development and Health, Southampton, United Kingdom; Data Science of a population-based birth cohort (Manchester Asthma and Allergy Institute, South Kensington Campus, Imperial College London, London, Study—MAAS), we investigated the value of FE V /FVC, BDR and FeNO United Kingdom; Acclarogen Ltd, St John’s Innovation Centre, Cam- 8 9 in diagnosing asthma in children. bridge, United Kingdom; AstraZeneca R&D, Mölndal, Sweden; BioSci Method: Using validated questionnaires we assessed study par- Consulting, Maasmechelen, Belgium; Dept of Respiratory Medicine, ticipants at age 16  years. Current asthma was defined as all three Academic Medical Centre, University of Amsterdam, Amsterdam, The of: (1) doctor-diagnosed asthma ever, (2) wheezing in the previous Netherlands 12 months and (3) current use of asthma treatment. We assigned chil- Correspondence: Andrew J. Simpson - Andrew.Simpson-2@Manchester. dren negative to all three features as non-asthmatic controls. Using ac.uk ATS/ERS guidelines, we measured spirometry and FeNO (NIOX chemi- Clinical and Translational Allergy 2017, 7(Suppl 2):O04 luminescence analyser; Sweden). BDR was considered positive if FEV increased by > 12% following administration of 400 mg of salbutamol. Introduction: Individuals with severe asthma may remain uncon- PEFR variability was not measured. To test the diagnostic algorithms trolled and exacerbation-prone despite intensive guideline-directed simulating the clinic situation, we selected only children reporting treatment, and management options are limited. The concept of treat- recent symptoms of wheeze, cough or breathlessness who were not able traits, based on the identification of treatable disease-associated on regular inhaled corticosteroids (ICS). characteristics, may thus be a particularly useful framework in this Results: Of the 630 MAAS children with full data available, 163 context. In the Unbiased Biomarkers for the Prediction of Respiratory reported recent symptoms, but were not using regular ICS; 34 of Disease Outcomes (U-BIOPRED) project we have recruited individuals these met our definition of current asthma, with 55 as non-asth- with severe asthma (SA) under specialist care, and controls with mild matic controls. In the multivariable logistic regression analysis, to moderate asthma (MMA). increasing FeNO was associated with an increased risk of asthma Aim: To identify and quantify treatable traits within the U-BIOPRED (OR 1.02, 95% CI 1.01–1.04, p  =  0.006), with a trend for FE V /FVC adult asthma cohorts. ratio (OR 0.95, 95% CI 0.87–1.02, p  =  0.17), and no association for Method: We defined criteria for treatable traits based on Agusti (Eur BDR (p  =  0.94). The proportion of those with each combination of Respir J, 2016) and identified prevalence rates within the U-BIOPRED positive tests is show as a Venn diagram (Figure  1). Of 58 children database. Chi Square tests were used to examine differences in fre - with three negative tests, 29.3% had current asthma, accounting quency between individuals with SA and MMA for 50% of those with asthma. Only 5.9% of those with asthma were Results: Data from 509 individuals with asthma were included in the positive to all three tests. analysis; 421 with SA and 88 with MMA. Twenty-nine treatable traits were Conclusion: Applying 3 tests of lung function to children with symp- identified, including 13 pulmonary, 13 extra-pulmonary and three behav - toms and a diagnosis of asthma failed to detect 50% of asthma cases. ioral traits. Pulmonary treatable traits such as airflow limitation (SA 50% Proposed algorithms for the diagnosis of asthma in symptomatic chil- vs. MMA 6%, P  <  0.001), reversibility (SA 58% vs. MMA 39%, P =  0.002), dren need to be tested prospectively. eosinophilia (SA 54% vs. MMA 43%, P = 0.067), exercise-induced asthma Keywords: Asthma, Diagnosis, FeNO, Lung Function, Children (SA 77% vs. MMA 54%, P  =  0.002), allergic rhinitis (SA 47% vs. MMA 44%, P = 0.641), cough (SA 63% vs. MMA 19%, P < 0.001) and bronchitis (SA 51% Vs. MMA 16%, P  =  0.000) were highly prevalent in the asthma cohorts, and typically more common in SA versus MMA. The most com- mon extra-pulmonary treatable traits were; atopy (SA 74% vs. MMA 92%, P < 0.001), obesity (SA 30% vs. MMA 38%, P = 0.178), reflux (SA 36% vs. MMA 11%, P  <  0.001), hypertension (SA 25% vs. MMA 9%, P  =  0.001) and obstructive sleep apnea (SA 26% vs. MMA 11%, P  =  0.003). Behav- ioral traits included low medication adherence (SA 39% vs. MMA 52%, P = 0.031) and smoking (SA 8%, smokers not eligible in MMA cohort). Par- ticipants with SA had mean (SD) 8 ± (2), and MMA 5 ± (2) treatable traits. Conclusion: We have applied a new approach for the characterisation of severe asthma, based on treatable traits. In general these traits were found more commonly in severe than non-severe asthma; the very high prevalence of many of these traits suggests that there are poten- tial targets for treatment even in such severe patients, and supports the need for specialist management. Keywords: Phenotypes, Treatable Traits Figure 2 Venn diagram showing number of children with symptoms (n163) who were positive for each combination of tests ORAL ABSTRACT SESSION 2—Molecular mechanisms O05 O04 Discovery, development and validation of blood‑based RNA Treatable traits in the European U‑BIOPRED adult severe asthma biomarker panels to predict the late‑phase asthmatic response 1 1 2 1 1 cohort S. J. Tebbutt , A. Singh , C. P. Shannon , Y. W. Kim , C. X. Yang , G. M. 1 2 3 4 3 1 4 3 Andrew J. Simpson , Dominick E. Sha w , Ana R. Sousa , Louise J. Fleming , Gauvreau , J. M. Fitzgerald , L. P. Boulet , P. M. O’Byrne 5 6 7 8 1 2 Graham Roberts , Ioannis Pandis , Aruna T. Bansal , Julie C orfield , Scott University of British Columbia, Vancouver, Canada; PROOF Centre 9 5 4 10 3 Wagers , Ratko Djukanovic , Kian Fan Chung , Peter J. Sterk , Jorgen of Excellence, Vancouver, Canada; McMaster University, Hamilton, 1 1 4 Vestbo , Stephen J. Fowler Canada; Université Laval, Québec City, Canada Division of Infection, Immunity and Respiratory Medicine, School Correspondence: Scott J. Tebbutt - scott.tebbutt@hli.ubc.ca of Biological Sciences, The University of Manchester and University Clinical and Translational Allergy 2017, 7(Suppl 2):O05 Clin Transl Allergy 2017, 7(Suppl 2):14 Page 3 of 17 Introduction: We have previously demonstrated significant molecu- Measurements of dynamic resistance were performed using a Flex- lar changes in the blood between mild allergic asthmatic individuals ivent system (Scireq, Montreal, Canada). After induction of anaesthesia who develop isolated early responses (early responders, ERs) com- with an intrperitoneal injection of ‘Hypnorm’ (0.315 mg fentanyl; 10 mg pared to those who also develop late-phase responses (dual respond- fluanisone) and midazolam (5  mg) in water, at a dose of 0.1  ml/10  g, ers, DRs) after allergen inhalation challenge. Identifying individuals mice were tracheostomized and connected to the flexivent ventilator likely to develop dual responses may aid in the screening of subjects via an 18-guage needle. Mice were ventilated. Changes in resistance for clinical trials that test drugs for the attenuation of the late-phase were measured in response to increasing concentrations of nebulized asthmatic response, which shares hallmark features of chronic disease. methacholine from 0 to 300  mg/ml. All measurements were made in The objective of this study was to develop blood-based biomarker the morning. −/− panels that could identify asthmatic individuals with high probability Results: Rev-erb alpha mice exhibit significantly (p  <  0.01) greater of developing a late-phase response. AHR to increasing concentrations of methacholine compared to their Method: The discovery cohort consisted of 36 mild asthmatic subjects wildtype and heterozygote litter-mate controls (Figure 1). −/− (15 ERs and 21 DRs) and a validation cohort consisted of 45 mild asth- Conclusion: Rev-erb alpha mice retain circadian rhythmicity yet matic subjects (9 ERs and 36 DRs). Blood samples were collected prior demonstrate increased AHR to methacholine (as is seen in human to allergen challenge. Following RNA extraction, total RNA (globin-/ asthma). REV-ERB alpha may play an important role in the pathogen- ribo-depleted) was sequenced using an Illumina HiSeq 2000 as 100 esis of asthma. base paired-end reads. Both genome-guided datasets (UCSC genes, Next steps: UCSC gene-isoforms, and Ensembl) and de novo assembled transcripts using the Trinity software were constructed. Top-ranked biomarker −/− 1. Investigate a time of day difference in AHR in Rev -erb alpha candidates were transferred to the high-precision and clinically- mice approved NanoString nCounter platform. Final biomarker panels were −/− 2. Expose Rev-erb alpha mice to the house dust mite model of identified, and statistical algorithms were locked down prior to testing asthma in the validated cohort. 3. Employ REV-ERB alpha ligands to ‘modify’ the phenotype Results: Predictive biomarker panels had classification performance (based on the area under the receiver operating curve, AUC) rang- Keywords: Airway Hyperresponsiveness, Rev-Erb Alpha, NR1D1, Clock ing between 60 and 70% in the discovery cohort. 87 transcripts Gene identified on the RNA-Seq platform were transferred to NanoString Elements assay chemistry. The transcripts were split with respect to their dataset of origin and tested in the validation cohort. The UCSC gene-isoforms and Trinity biomarker panels had AUCs of 68% and 72%, respectively. The biomarker transcripts were enriched for bio- logical pathways such as NF-ĸB signaling (up-regulated in DRs) and apoptosis, decoy receptors and formyl peptide receptors (down-reg- ulated in DRs). Conclusion: RNA transcript biomarker panels in the blood were successful at predicting asthmatic subjects likely to develop dual responses upon allergen inhalation challenge. Pathways and networks represented by these biomarker panels may reveal new avenues for therapeutic targeting of the pathobiology involved in chronic asthma. Keywords: Predictive Biomarkers, RNA-Seq, NanoString Elements, Sta- tistical Algorithms O06 The clock Gene REV‑ERBalpha regulates airway Figure 3 Airway Hypersensitivity to Methacholine hyperresponsiveness: implications for asthma H. J. Durrington, N. Begley, A. Loudon, D. W. Ray University of Manchester, Manchester, United Kingdom Correspondence: Hannah J. Durrington - hannah.durrington@manches- O07 ter.ac.uk Asthma biomarkers: methylation and correlation with functional Clinical and Translational Allergy 2017, 7(Suppl 2):O06 parameters 1 1 1 2 Selene Baos , Lucía Cr emades , David Calzada , Carlos Lahoz , Blanca Introduction: Asthma is an inflammatory disease with a strong circa- Cárdaba 1 2 dian signature. Symptoms of asthma are worse overnight and in the IIS-Fundación Jiménez Díaz-UAM, Madrid, Spain; IIS-Fundación Jiménez early hours of the morning. Measurements of peak expiratory flow Díaz-UAM; Ciber de Enfermedades Respiratorias, CIBERES, Madrid, Spain rate and forced expiratory volume in 1  s, are lower in early morn- Correspondence: Selene Baos - selene.baos@fjd.es ing compared to afternoon. It is likely that circadian biology plays Clinical and Translational Allergy 2017, 7(Suppl 2):O07 a role in the pathogenesis of asthma. A better understanding of the circadian nature of asthma may lead to better treatments through Introduction: In a previous study (submitted to publish) we defined chronotherapy (taking medication at the most beneficial time of day). specific genes related with asthma and allergic diseases, by study - Clock genes control circadian rhythms within every cell in the body. ing the gene-expression of 94 genes in a population composed by 4 The clock gene, REV-ERB alpha (NR1D1), connects the ‘core’ clock and groups of subjects: healthy control, nonallergic asthmatic, asthmatic the immune system. We employed a REV-ERB alpha global knockout allergic and nonasthmatic allergic patients. The analysis of differen- −/− mouse (rev-erb alpha , these mice retain circadian rhythmicity, but tial gene-expression between control and patients with nonallergic show loss of temporal variation of innate immunity) to investigate the asthma revealed a set of statistically relevant genes mainly associated effect of REV-ERB alpha on airway hyper-responsiveness (AHR). AHR is with the disease’s severity: IL10, MSR1, PHLDA1, SERPINB2, CHI3L1, a key pathophysiological feature of asthma. IL8, and PI3. In this project we analyzed if methylation takes part in Method: All experimental procedures were carried out in accordance the regulation of the gene-expression of these potential asthma bio- −/− with the Animals (Scientific Procedures) Act,1986. Rev-erb alpha markers and their correlation with protein expression and functional mice were provided by Ueli Schibler (University of Geneva). parameters (%FVE1,  %FVC and  %PBD). Clin Transl Allergy 2017, 7(Suppl 2):14 Page 4 of 17 Method: DNA extracted from PBMCs of control and nonallergic O09 asthma subjects was treated with sodium bisulfite and amplified by Hyaluronan‑induced lipid mediators influence antiviral PCR with primers designed to amplify CpG islands near the promotor and antibacterial immunity in severe asthmatics 1 2 2 region of the most significant genes: MSR1 (5 CpG islands), SERPINB2 Milena Sokolowska , Li-Yuan Chen, Yueqin Liu , Asuncion 2 2 2 3 (5), PHLDA1 (35), CHI3L1 (8) and PI3 (5). The methylation analysis was Martinez-Anton , Carolea Logun , Sara Alsaaty , Rosemarie Cuento , 2 2 4 5 done with the Sequenom EpiTYPER approach. Protein quantification Rongman Cai , Junfeng Sun , Oswald Quehenberger , Aaron Armando , 5 3 2 was determined by ELISA or Western Blot. Statistical analyses were Edward Dennis , Stewart Levine , James Shelhamer performed with Graph-Pad program. Critical Care Medicine Department, Clinical Center, NIH; Swiss Institute Results: The methylation analysis showed statistically significant dif- of Allergy and Asthma Research, University of Zurich; CK-CARE, Davos, ferences between groups in 4 genes: MSR1, PHLDA1, CHI3L1 and PI3. Switzerland; Critical Care Medicine Department, Clinical Center, NIH, The results of the methylation correlated with the gene-expression, Bethesda, Maryland, United States; Laboratory of Asthma and Lung the protein levels and respiratory parameters, specifically in MSR1, Inflammation, Cardiovascular and Pulmonary Branch, National Heart, CHI3L1 and PI3. Lung and Blood Institute, NIH, Bethesda, Maryland, United States; Conclusion: A possible regulatory mechanism of molecular biomark- Department of Medicine, Department of Pharmacology, San Diego, ers of asthma has been defined, being methylation a possible key fac - California, United States; Department of Chemistry and Biochemistry, tor for the differential gene-expression of asthma patients. San Diego, California, United States Keywords: DNA Methylation, Biomarkers, Epigenetics Correspondence: Milena Sokolowska - milena.sokolowska@siaf.uzh.ch Clinical and Translational Allergy 2017, 7(Suppl 2):O09 O08 Critical role for Arginine metabolism in a combined Th2 and Th17 Introduction: Hyaluronan (HA) is the major glycosaminoglycan in the airway inflammation in the house dust mite model extracellular matrix involved in the pathogenesis of asthma and other Kewal Asosingh, Chris Lauruschkat, Kimberly Queisser, Nicholas Wanner, chronic inflammatory diseases. During inflammation in asthma there is Kelly Weiss, Weiling Xu, Serpil Erzurum an increased breakdown of HA, resulting in the local and systemic accu- Cleveland Clinic, Cleveland, Ohio, United States mulation of low molecular weight (LMW) HA. Eicosanoids, derived from Correspondence: Kewal Asosingh - asosink@ccf.org cytosolic phospholipase A group IVA (cPLA alpha) activation, are potent 2 2 Clinical and Translational Allergy 2017, 7(Suppl 2):O08 lipid mediators also attributed to acute and chronic inflammation. Method: We investigated the effect of LMW HA on the lipidomic pro - file and global gene expression and their functional interactions in Introduction: Inducible nitric oxide synthase (iNOS) and arginase-2 peripheral blood mononuclear cells of patients with mild-to-moderate (ARG2) share a common substrate arginine. We recently showed (n = 7) and severe asthma (n = 6) as compared to controls (n = 6). that both enzymes are highly expressed in asthmatic airway epi- Results: We found that LMW HA increased production of 68 unique thelial cells where ARG2 plays a critical role in the regulation of lipid species, among which PGE , PGB, PGD, 15-HETE, TxB , 11(12)- 2 2 2 2 mitochondrial bioenergetic driven Th2 response, as shown by the EET, 14(15)-EET, 13-HOTrE(y) and 16(17)-EpDPE were significantly development of severe eosinophilic airway inflammation in mice upregulated only in severe asthmatics. We also performed a genome- deficient for ARG-2 (ARG-2 KO). The role of iNOS and ARG2 in a com- wide expression analysis of LMW HA signaling, confirming its highly bined Th2 and Th17 response is unknown. Here we hypothesize immunostimulatory potential. However, in severe asthmatics the LMW that Th17 airway inflammation is also regulated by ARG2 driven HA-induced global gene expression profile showed a comprehen- metabolism. sive impairment in interferon signaling, cell apoptosis and cell move- Method: Commercially available Wildtype (WT) and iNOS knockout ment, leading to diminished antiviral and antibacterial responses. We (KO) were used. ARG2 KO was donated to us and ARG2iNOS double KO confirmed these findings at the protein level, finding that LMW HA- (dKO) strain was generated in house. Standard house dust mite extract induced production of IL-12 p40, CXCL10, CXCL11 and CCL8 in severe (HDME) mouse model of Th2/Th17 asthma endotype was used. Air- asthmatics was markedly reduced. Importantly, upon cPLA alpha inhi- way inflammation was quantified by analysis of inflammatory cells in bition, there was a significant decrease in lipid mediator production bronchoalveolar lavage fluid (BALF). Airway angiogenic remodeling accompanied by a significant increase in IL-12 p40 and CXCL9 protein was measured by quantification of lung microvessel density on tis- expressions in each phenotype. sue sections stained for the endothelial marker von Willebrand Factor. Conclusion: In summary, we demonstrated here that fragmented hya- Epithelial and immune cell cytokines were measured by ELISA or flow luronan increased production of several unique lipid species in severe cytometric multiplex assay. asthmatics. Moreover, through analysis of the whole genome pro- Results: HDME induced airway inflammation in all genotypes, but file, captured simultaneously with the lipidomic profile, we observed was the highest in ARG2 KO as showed by increased number inflam- decreases in antiviral gene and protein expression upon LMW HA treat- matory cells in the BALF. Both eosinophilic (Th2) and neutrophilic ment in severe asthmatics, which was partially reversed upon inhibition (Th17) inflammation peaked in ARG2 KO mice but were attenuated of lipid production. Therefore, our current findings provide new evidence in ARG2iNOS dKO animals. Airway inflammation was similar among on the connection between extracellular matrix, global lipid mediator HDME exposed WT, iNOS KO and ARG2iNOS dKO mice. Angiogenic production and decreased antiviral responses in severe asthma. airway remodeling was also highest in ARG2 KO mice and comparable Keywords: Severe Asthma, Eicosanoid, Virus, Lipidomics, among WT, iNOS KO and ARG2iNOS dKO genotypes. Airway epithelial Transcriptomics or immune cell-derived Th2 cytokines IL-5 and eotaxin-2 and, Th17 cytokine IL-17 were maximal in lungs of HDME exposed ARG2 KO mice, but decreased in ARG2iNOS dKO. POSTER DISCUSSION SESSION 1—Epidemiology Conclusion: The data show that ARG2 deficiency induces a severe combined Th2 and Th17 airway inflammation. Deletion of iNOS in P01 addition to ARG2 inhibited disease severity, but iNOS deletion alone, Fungal communities in house dust associated with the had no effect, indicating that ARG2 is the most critical of the two argi- development of Atopic Dermatitis in infant nine consuming enzymes in the regulation of inflammation and angi- Kilyong Choi ogenesis in asthma. Overall, the results expand our previous findings Seoul Medical Center, Seoul, South Korea by demonstrating that in addition to Th2, Th17 endotype of airway Correspondence: Kilyong Choi - bestchoi9494@gmail.com inflammation is also critically regulated by arginine metabolism Clinical and Translational Allergy 2017, 7(Suppl 2):P01 Keywords: Arginine, Metabolism, Th2, Th17, Mitochondria Clin Transl Allergy 2017, 7(Suppl 2):14 Page 5 of 17 Introduction: In our epidemiologic study, the effect of mold exposure Table 2 List of fungal genera and their relative abundance was associated with the exacerbation or the development of atopic der- (%) in house dust samples from two groups matitis (AD). Maternal exposure to mold during the pregnant periods or exposure to mold in early life particularly have an impact on the increased Taxon Status risk of AD development. However, its specific mechanism still remains Phylum Genus Health AD (n = 20) p Value unclear. In addition, dust is a repository and concentrator of many chemi- (n = 20) cal and biological agents including fungi. The aims of this study were to identify the fungal microbiota (mycobiota) in house dust, and to investi- Ascomycota Alternaria 7.9104 12.8797 0.079 gate whether the mycobiota is associated with the development of AD. Ascomycota Aspergillus 4.1237 5.1118 0.705 Ascomycota Aureobasidium 5.8441 5.0303 0.85 Table  1 Library coverage estimations and  sequence diver‑ Ascomycota Candida 0.7504 1.0536 0.725 sity of fungal ITS genes pyrosequencing Ascomycota Cladosporium 7.8394 7.6885 0.725 ID Valid OTUs Chao1 Shannon Simpson Goods Ascomycota Cyphellophora 0.5648 0.6081 0.022 reads Lib. Ascomycota Exophiala 0.3533 0.8299 0.957 Coverage Ascomycota Fusarium 0.6404 1.3015 0.685 S13-033-C 9475 474 769.5882 3.619653 0.114784 0.978786 Ascomycota Leptosphaer- 1.4300 0.8881 0.291 S12-073-C 8588 456 567.125 4.541252 0.02198 0.985212 ulina S12-067-C 10169 611 731.3051 4.629529 0.024318 0.983381 Ascomycota Meyerozyma 0.0367 1.7910 0.989 A13-033-C 9719 377 455.9153 3.690041 0.090025 0.99002 Ascomycota Nigrospora 0.7219 0.5520 0.344 A12-080-C-22 7111 315 404.7581 3.541059 0.09324 0.985094 Ascomycota Penicillium 2.7624 1.3681 0.978 A12-072-C 7899 439 757.3333 3.991715 0.051364 0.97582 Ascomycota Peyronellaea 1.6187 2.7516 0.818 A12-054-C 9467 343 574.6667 3.859098 0.05143 0.985212 Ascomycota Phoma 7.1393 6.3727 1 Y10-533-C 10273 520 775.6081 4.314013 0.038003 0.981018 Ascomycota Pleospora 0.4702 0.5722 0.487 C12-044-C 8788 382 518.1224 4.288241 0.030933 0.9868 Ascomycota Saccharomyces 2.9811 1.0517 0.176 Y12-016-C 11603 576 952.9615 4.045346 0.052539 0.979057 Basidiomycota Cryptococcus 1.8277 2.1603 0.725 A13-047-C 9386 517 740.9474 4.419917 0.037477 0.98029 Basidiomycota Malassezia 10.8815 4.4114 0.005 C13-065-C 11197 469 599.013 4.253351 0.033662 0.987318 A10-581-C 11167 530 693.4 4.24386 0.047244 0.984597 Basidiomycota Malassezi- 0.8520 1.2991 0.626 aceae_uc A10-609-C 12620 725 1126.857 4.347734 0.035431 0.976941 Basidiomycota Pleurotus 3.2915 0.0260 0.005 A12-037-C 11089 828 1181.979 4.64697 0.035575 0.970962 Basidiomycota Rhodotorula 1.5704 0.7741 0.725 A13-069-C 12210 764 1144.971 4.907156 0.02687 0.976904 Basidiomycota Trichosporon 4.1956 0.3016 0.003 C13-067-C 10215 570 733.8298 4.774492 0.021033 0.98277 S12-072-C 6413 317 375.2632 3.910256 0.058516 0.987213 Y12-044-C 12008 638 949.3542 4.230534 0.070634 0.979597 A12-071-C 4753 544 746.4118 5.021128 0.018621 0.960867 Method: To identify the mycobiota in house dust, DNA pyrosequenc- Y12-051-C 7282 329 416.3061 4.209385 0.030832 0.987229 ing was performed targeting the internal transcribed spacer (ITS) A10-575-C 9011 608 787.1 4.68377 0.025285 0.977916 region. Here, we investigated the composition of fungal communities in the dust samples from houses of 20 healthy and 20 AD infants diag- A12-074-C 10195 410 549.2642 3.913959 0.060137 0.988033 nosed at 6 and/or 12 months. A13-023-C 10885 366 425.7143 4.232292 0.035282 0.992926 Results: A total of 7 phyla and 399 fungal genera were distinguished A13-016-C 11250 573 778.8875 4.588823 0.026887 0.983822 in house dust. Fungal alpha diversity was similar between two groups. However, our results show that the different fungal communities C12-042-C 8323 341 639.5294 3.248262 0.102067 0.978974 between healthy and AD groups. The genera Alternaria, Aspergillus, A13-048-C-21 10255 504 647.3544 4.15016 0.049551 0.985275 Fusarium and Candida belonging to Ascomycota and known as the S12-056-C 11210 940 1505.511 4.965526 0.03022 0.964585 common allergen were higher abundant in dust samples for infants S12-052-C 10900 578 787 3.604678 0.118101 0.980734 with AD. On the other hand, Malassezia, Pleuotus and Trichosporon belonging to Basidiomycota were less abundantin dust samples for A12-051-C 11160 688 1004.469 4.548595 0.0311 0.977867 infants with AD. A12-036-C 11206 492 711.6133 3.816188 0.086481 0.983759 Conclusion: House dust from infants with AD show higher abundance C13-033-C 11982 562 863.8889 4.367747 0.030685 0.982557 of specific fungi known as allergens, which suggests that different Y13-018-C 11885 496 632.2805 4.114662 0.045829 0.987379 fungal exposure from the dust may link to the development of AD in infancy. A10-596-C 8604 438 726.0159 3.686982 0.075458 0.977801 Keywords: House Dust, Atopic Dermatitis, Infant, Fungal Communities S13-025-C 9653 503 786.5156 4.473125 0.025739 0.980213 S13-029-C 10485 580 815.6915 4.305426 0.03932 0.979876 P02 C14-006-C 11607 633 941.9326 4.389317 0.04185 0.979754 Cluster analysis in Bulgarian children with asthma 1 1 1 C14-009-C 10267 528 823.5882 4.297914 0.050246 0.980423 Snezhina Lazova , Penka Perenovska , Dimitrinka Miteva , Stamatios 2 1 Priftis , Guergana Petrova S12-090-C 10219 480 665.2442 4.017626 0.053966 0.982484 Medical University, Pediatric clinic, UMHAT Alexandrovska, Sofia, Bulgaria; S13-045-C 10677 421 560.2344 3.541324 0.118609 0.98745 Medical University, Sofia, Bulgaria Total 401206 20865 Correspondence: Snezhina Lazova - snejina@lazova.com Clinical and Translational Allergy 2017, 7(Suppl 2):P02 Clin Transl Allergy 2017, 7(Suppl 2):14 Page 6 of 17 Introduction: Asthma is a complex heterogeneous disease that likely Patients with both scoliosis and asthma, should be managed as severe comprises several distinct disease phenotypes. Usually atopy presence asthma patients. Evaluation of their PFT is essential in their manage- or any allergic diseases could interfere and obstacle the control of ment plan for early intervention should not only restriction type defi- asthma. Clustering approach has been used to classify heterogeneous ciency is noted asthma population into distinct phenotypes. Keywords: Scoliosis, Severe Asthma, FEV1 Method: For a period of 1  year we evaluated medical history data of 110 children with asthma aged 3 to 17  years. For all children we per- Acknowledgements: This work was supported by a grant from the formed pulmonary function tests, nasal smears for eosinophil counts, Medical University of Sofia (Council of Medical Science, project no. drew blood for IgE against inhalation and food allergies antibodies 513/2016, grant no. 65/2016). detection and ACQ. IgE were detected with the predesigned kit Euro- linePediatric and total IgE with Immunocap. Differences in clinical P04 indices including ΔFEV1(pre- and post-bronchodilator) and nasal and The cases of asthma in Japan have been expanding or not. Study blood biomarkers at enrollment among clusters were analyzed. on the tendency of asthma from the official statistical reports Results: Five distinct phenotypes were determined. Cluster 1 (n = 13): Takashi Kumae (non-atopic) the lowest IgE level, very low ACQ, median age of diag- Research Institute, Tokyo University of Agriculture, Tokyo, Japan nosis and IgE levels. Cluster 2 (n  =  45): (mixed) the highest BMI with Correspondence: Takashi Kumae - r071018@kanto-gakuin.ac.jp the latest age of diagnosis and high ACQ and BDR levels. Cluster 3 Clinical and Translational Allergy 2017, 7(Suppl 2):P04 (n = 33) (atopic) early diagnosis, highest BDR, highest ACQ score, high- est total and specific IgE levels among the clusters. Cluster 4 (n  = 12): Introduction: For consideration and discussion of data obtained from (atopic) the highest Immunocap result, relatedly high BMI and IgE with a survey on asthma, two official statistical reports are available in median ACQ score among clusters. Cluster 5 (n = 7): (non-atopic) the Japan. The fundamental data in these reports are statistically analyzed earliest age for diagnosis, with the lowest BMI, the lowest ACQ score, and compared to clarify the tendency of asthma in Japan. and Immunocap result, with high BDR and median level of IgE among Method: One is annual report of school health statistics (SHS) from clusters. Ministry of Education, Culture, Sports, Science and Technology which The patients with severe asthma were over 65% of the patients in clus- has competence to survey the school age population including dis- ter 3. Only one severe asthma patient was in cluster 2, and none in the eases and related health problems.  The incidence of asthma of SHS other three clusters. from 1995 to 2015 in 5 to 17 years old are used in this report. The other Conclusion: We identified asthma phenotypes in Bulgarian children is the patient survey by Ministry of health, Labour and Welfare. The according IgE levels, ACQ score, BDR and age of diagnosis. Almost all patient survey covers all generations and diseases but the survey car- of the severe asthma patients were in cluster 3 – with early diagnosis ried on every third year and numbers of patients are summarized by and highest IgE but not highest Immunocap results. 5 years old. Numbers of asthma case from 1996 to 2014 are extracted Keywords: Atopy, IgE, ACQ, Severe Asthma and adjusted to the corresponding age population as per 1,000 capita in this report. P03 Results: According to SHS, the incidence of asthma was most frequent Lung function parameters in asthma, severe asthma and scoliosis in 6  years old and statistically significant positive correlations were in children observed between the incidence and a lapse of years in all ages. Aver- 1 2 1 2 Snezhina Lazova, Vassil Yablanski , Penka P erenovska, Evgeni Vlaev , age slope of regression lines in the 5 to 9 years old was 0.10 in female 1 3 1 Hristina Rafailova , Stamatios Priftis , Guergana Petrova and 0.16 in male. This analysis means that the incidence may be keep Medical University, Pediatric cilinic, UMHAT Alexandrovska, Sofia, Bul- increase over 0.1% in annual in the near future. From the patient sur- 2 3 garia; Orthopedics department, Tokuda Hospital, Sofia, Bulgaria; Medical vey, asthma cases were most numerous in the 0 to 4  years old group University, Sofia, Bulgaria and the 5 to 9  years old group was next. Asthma cases increased in Correspondence: Snezhina Lazova - snejina@lazova.com elderly and the level of the 70 to 74 years old group was almost same Clinical and Translational Allergy 2017, 7(Suppl 2):P03 level of the 10 to 14 years old group. Contrary to SHS, statistically sig- nificant negative correlations were observed between the asthma cases and a lapse of years in the age groups, 15 to 19 (p  <  0.01), 20 Introduction: Pulmonary function testing (PFT ) is of great importance to 24 (p < 0.05) and 25 to 29 (p < 0.05) years old in male. The analysis in the evaluation of lung function. Spirometry is simple, noninvasive, of the patient survey suggests that the asthma has decreased in these and has been the most commonly used technique in children. Scolio- two decades. sis is the most common abnormality of the spine with direct effects Conclusion: Contradictory results in younger generation are obtained on the thoracic cage. Scoliosis has generally been associated with the by the analysis of the two official statistical reports. The patient sur - development of restrictive lung disease. vey is based on actual number of persons had a medical examina- Objectives: To evaluate the PFT data of children with scoliosis and to tion in medical facilities, on the other hand, SHS is carried out using compare them to children with asthma. the questionnaire. The data of SHS is seemed to be influenced by the Method: After obtaining signed informed consent from the parents anamnesis. Especially in lower age children, SHS data are also possibly we performed PFT in 50 children aged 5 to 17 years. The children were influenced by the levels of interests and anxieties for asthma in their divided into four groups – 10 children with scoliosis without asthma protectors. (Sc),10 children with scoliosis and asthma (AS), 10 children with mod- Keywords: Annual Report Of School Health Statistics, Patient Survey, erate asthma (BA), 10 children with severe asthma (SA) and 10 healthy Statistical Analysis children (HC). Results: The four groups have similar age (p  =  0.079), sex (p  =  0.19) P05 and FVC/FEV1 (p  =  0.403) distribution. The results for FVC  %pred, Patients’ perspectives of the impact of severe asthma on sex, FEV1%predand MMEF 25/75%pred were significantly different with intimacy and relationships: too taboo to talk about? p = 0.009 (Sc vs AS), p = 0.002 (SA vs BA) and p = 0.001 (AS vs BA), all 4 1 2 3 L. J. Holmes, J. Yorke , D. M. Ryan groups had lower results compared to HC (p < 0.05). The same param- University Hospital of South Manchester & University of Manchester, eters were similar when comparing Sc with BA and SA with AS. The Manchester, United Kingdom; University of Manchester, Manchester, lowest FVC %pred was in the AS group, even lower than SA (p = 0.06). United Kingdom; University Hospital of South Manchester, Manchester, The children with SA demonstrated the lowest MMEF 25/75%. The United Kingdom FEV1% pred were higher in healthy group while children with scoliosis Correspondence: Leanne-Jo Holmes - leanne.holmes1@ntlworld.com and asthma have comparable lower values. Clinical and Translational Allergy 2017, 7(Suppl 2):P05 Conclusion: The children with scoliosis demonstrated diminished expiratory flow rates, while the FEV1/FVC ratio is within normal ranges. Clin Transl Allergy 2017, 7(Suppl 2):14 Page 7 of 17 Introduction: Patients with severe asthma experience unpredictable measured by ELISA. Spot urine samples were measured for urinary daily symptoms and an intense treatment regimen that can impact Leukotriene E4 (uLTE4) levels by enzyme immunoassay. upon physical, emotional well-being and quality of life. Sexual func- Results: Two hundred and nine data collecting forms with the bio- tion is a basic human requirement, contributing to quality of life; yet assay results were available for analysis in this report. The mean age there is a dearth of research exploring relationships and intimacy in of the participants was 26  months with boys making up 62.5%. First people with severe asthma. The overall aim was to explore the impact wheeze episode accounted for 40.6 percent. Seventy-five percent was of severe asthma on patient’s experiences of intimacy and relation- atopic with mono-sensitization (29.3%). The highest prevalence of ships, establish their information needs and the role of the healthcare sensitization was egg (45.6%), followed by cow’s milk (45.3%). The high professional. median urine leukotriene E4 (uLTE4) levels of 226  pg/ml and 956  pg/ Method: A qualitative study using semi-structured interviews with mg creatinine was documented. The prevalence of vitamin D insuffi- patients receiving treatment for severe asthma at a dedicated severe ciency, defined by the serum 25OHD levels of less than 30 ng/ml, was asthma clinic was undertaken. Interviews were audio recorded and detected in 22.3%. transcribed verbatim. Transcripts were verified then analysed for emer - Conclusion: Majority of preschool children with acute wheezing gent themes which were then categorised into super-ordinate themes admitted in the hospital had sensitization with egg and cow’s milk. and sub themes. One-fourth of them had vitamin D insufficiency and associated with Results: All nine participants had a proven diagnosis of severe high serum level of uLTE4. asthma. A higher proportion were female n  =  6 (66.6%) compared Keywords: Allergic Sensitization, Acute Wheezing to male n = 3(33.3%). The mean age was 45 years, range 34–59 years. The majority of participants 8(88.9%) were married, with 1(11.1%) P07 co-habiting. Cluster analysis of patients with moderate to severe uncontrolled The analysis of interviews provided complex, detailed and novel bronchial asthma insights into patients’ perspectives on how living with severe asthma 1 1 1 Vania Youroukova , Denitsa Dimitrova, Yanina Slavova , Spaska impacts upon intimacy and spousal relationships. Four super-ordinate 2 Lesichkova themes emerged: 1 Medical Faculty of Medical University of Sofia; Clinical center of pulmo - nary diseases; SHATPD “St. Sofia”, Sofia, Bulgaria; Medical Faculty of Medi- cal University of Sofia, Department of Clinical Immunology, University (i) ‘Physical intimacy’ including disclosure of physical limitations of Hospital Alexandrovska, Sofia, Bulgaria severe asthma upon intimacy and sexual activity. Correspondence: Denitsa Dimitrova - ddimitrova87@abv.bg (ii) ‘Emotional intimacy’ the cyclical impact of the often negative Clinical and Translational Allergy 2017, 7(Suppl 2):P07 emotional struggle of living with severe asthma upon both rela- tionships and intimacy. (iii) ‘Image of self’ as participants divulged their battle with body Introduction: The aim of the study is to identify distinct subgroups of image and confusion in changing relationship roles. adult patients with uncontrolled moderate to severe bronchial asthma (iv) ‘The role of the healthcare professional’ patient identified infor - (BA) based on clinical and inflammatory characteristics. mation requirements within this area. Method: Adults with uncontrolled moderate to severe asthma (n  =  23) underwent a clinical assessment, spirometry and measure- Conclusion: The participants in this study have provided invaluable ment of fractional exhaled nitric oxide (FeNO). Total and specific serum and novel insights into the implications of living with severe asthma IgE levels, eosinophil count in peripheral blood and sputum were eval- upon a sensitive and rarely discussed topic. It is anticipated that these uated. K-mean cluster analysis was used to identify clinically relevant findings will serve to increase understanding and assist practitioners subgroups. to help patients to adopt positive strategies to improve their quality of Results: We have identified four distinct groups based on onset, atopic life within this area. status, FEV1/FVC ratio and FeNO. Cluster 1 patients (eosinophilic asthma) (n  =  6; 26.1%) were with late-onset asthma, predominantly non-atopic with deteriorated lung function and highest level of FeNO. P06 In this cluster were observed the highest blood and sputum eosino- Clinical characteristics and allergic sensitization of preschool phil counts (p = 0.01; p = 0.04) and more frequent exacerbations (≥ 1 children hospitalized with acute wheezing: a multi‑cohort study 1 2 exacerbation in previous 12 months) (p = 0.03). Cluster 2 patients (late- Sasawan Chinratanapisit , Khlongtip M atchimmadamrong , Jitladda 3 4 5 onset, atopic asthma) (n  =  7; 30.4%) were late-onset, mainly atopic Deerojanawong, Wissaroot Karoonboonyanan , Paskorn Sritipsukho with high total serum IgE levels, lower levels of FeNO and eosinophil Division of Allergy & Immunology, Department of Pediatrics, Bhumi- counts and deteriorated lung function. Cluster 3 patients (non-atopic bol Adulyadej Hospital, Bangkok, Thailand; Saraburi hospital, Saraburi, asthma) (n = 6; 26.1%) were late-onset, mainly non-atopic, with lower Thailand; Division of Respiratory disease and intensive care, Department levels of FeNO and eosinophil counts and well-preserved pulmonary of Pediatrics, Faculty of Medicine, Chulalongkorn University, Bangkok, function. Cluster 4 patients (early-onset, atopic asthma) (n = 4; 17.4%) Thailand; Department of Pediatrics, Bhumibol Adulyadej Hospital, Bang- were early-onset, atopic, with low levels of FeNO and preserved lung kok, Thailand; Center of Excellence in Applied Epidemiology, Thammasat function. In cluster 3 and 4 prevailed patients with high BMI (n = 7). University, Bangkok, Thailand Conclusion: Our current results suggests that uncontrolled asthma is Correspondence: Sasawan Chinratanapisit - sasawan2001@yahoo.com heterogeneous disease. Cluster identification will result in a personal- Clinical and Translational Allergy 2017, 7(Suppl 2):P06 ized approach for each patient and improvement of therapy response. The research was funded by GRANT project № 308/2015, contract Introduction: Background: Wheezing is a significant health problem №75/2015 to CMS, MU Sofia in Thailand, especially in the preschool age. Keywords: Clusters, Uncontrolled Adult Asthma, Inflammatory Objective: In this multi-center clinical research, clinical characteristics Characteristics and biomarkers were studied to determine the factors associated with persistent wheezing in the next three years among pediatric patients P08 with acute wheezing who were less than five years of age. Influence of respiratory viruses on the severity of bronchial Method: This prospective observational study recruited children, aged obstruction in preschool wheezing children 6 months to 5 years, with acute wheezing from the pediatric wards of 1 1 1 2 Iren Tzocheva , Snezhina Lazova , Snezhana Parina , Svetla Angelova , four hospitals from July 2014 to June 2015. Patients’ characteristics 2 1 Neli Korsun , Penka Perenovska and their family data were collected. Blood samples were collected for Pediatric clinic, University Hospital Alexandrovska, Medical University, detecting ImmunoCAP specific IgE against common food and inhal- Sofia, Bulgaria; National Centre of Infectious and Parasitic Diseases, ant allergens. Serum 25-hydroxyvitamin D (25OHD) concentration was Clin Transl Allergy 2017, 7(Suppl 2):14 Page 8 of 17 National Laboratory Influenza and ARD, Sofia, Bulgaria Results: In 2014 were 880 ED visits of 296 patients. 16 children were Correspondence: Snezhina Lazova - snejina@lazova.com included in the tele-monitoring study, 4 with severe asthma and 38 as Clinical and Translational Allergy 2017, 7(Suppl 2):P08 controls. Patients had male predominance [75% vs 71.1% in controls]. 2 (1 of the 4 severe asthma patients) vs 11 children in control group had more than 1 visit to ED in 2014. Odds-ratio to have an unsched- Introduction: Respiratory infections are associated with wheezing ill- uled ED visit was 0.1964 in tele-monitoring group [p 0.0483, 95% CI nesses at all ages and may also impact the development and sever- 0.039 – 0.9881]. Was higher for severe asthma patients compared with ity of asthma. Wheezing illnesses in young children are associated up non-severe with an odds–ratio 3.667 but non-significant [p 0.4040, to 95% with respiratory viral infections. In children with established 95% CI 0.1734 to 77.5559] asthma, viral respiratory tract infections play a key role in producing Conclusion: Tele-monitoring via virtual media can decrease the bur- acute exacerbations that may lead to hospital admission. The aim of den of unscheduled ED visits for asthmatic children, even in severe the present study is to investigate the relationship between various asthma patients. respiratory pathogen and severity of bronchial obstruction in hospital- Keywords: Child, Asthma, Virtual-Media, Monitoring, Education ized preschool children. Method: Nasopharyngeal secretions of 57 children aged 2 to 5  years P10 hospitalized with wheezing episode (Jan to Mar 2016) were examined Challenges in using hierarchical clustering to identify asthma for respiratory viruses (HMPV; RSV A/B; PIV 1, 2, 3; RV, AdV; Influenza subtypes: choosing the variables and variable transformation A/B) with Real Time PCR. 1 2 3 4 Matea Deliu, Tolga Yavuz , Matthew Sperrin , Umit M. Sahiner , Danielle Results: Respiratory viral infections was detected in 84% of the 5 6 7 8 Belgrave , Cansin Sackesen Sackesen , Adnan C ustovic , Ömer Kalayci patients. The predominant pathogen were RSV (52%), followed by Institute of Population Health, Health e-Research Centre, University influenza virus A (25%), human metapneumovirus (12.5%), rhinovirus of Manchester, Manchester, United Kingdom; Division of Pediatric (4%), parainfluenza (4%) and adenovirus (2.5%). In children with life- Allergy, GATA Military School of Medicine, Ankara, Turkey; Centre threatening bronchial obstruction with a need of prolonged oxygen for Health Informatics, Institute of Population Health, University of Man- therapy, RSV was the most commonly detected pathogen (42%). The chester, Manchester, United Kingdom; Division of Pediatric Allergy, presence of viral infection did not correlate with the duration of the Hacettepe University School of Medicine, Ankara, Turkey; Faculty systemic corticosteroid treatment. The median duration of reported of Medicine, Department of Medicine, Imperial College London, London, lung sounds (wheezing and crackles) was longer in the patients with 6 7 United Kingdom; Koç University Hospital, Istanbul, Turkey; Department detected virus infection without statistical significance (p = 0.06). of Paediatrics, Imperial College London, London, United Kingdom; Pedi- Conclusion: Respiratory viruses play a key role in the wheez- atric Allergy and Asthma Unit, Hacettepe University School of Medicine, ing illnesses in young children, predominantly infants. Our results Ankara, Turkey demonstrated that the RSV is an important pathogen in severe, life- Correspondence: Matea Deliu - matea.deliu-2@postgrad.manchester. threatening episodes in preschool age children as well. Contrary to ac.uk data presented in literature, human rhinovirus was detected in only 4% Clinical and Translational Allergy 2017, 7(Suppl 2):P10 of our patients following Influenza A and human metapneumovirus. Keywords: Preschool Wheezing, Respiratory Viruses, Asthma Exacer- bation, Severity Introduction: The use of unsupervised clustering has identified differ - ent subtypes of asthma. Choosing the variables to input into the clus- P09 tering algorithm is one of the important considerations. The majority Tele‑monitoring decreases unscheduled outpatient visits of previous studies selected variables based on expert advice, whilst in pediatric patients with severe asthma others used dimension reduction techniques such as principal compo- Mihai Craiu, Iustina Violeta Stan nent analysis (PCA). We aimed to compare the results of unsupervised INSMC Alessandrescu-Rusescu, Bucharest, Romania clustering when using raw variables, or variables transformed using Correspondence: Mihai Craiu - mcraiu@yahoo.com dimensionality reduction techniques. Clinical and Translational Allergy 2017, 7(Suppl 2):P09 Method: We performed our analysis on 613 asthmatics aged 6–23 years from Ankara, Turkey. We conducted extensive phenotyping and recorded 49 variables including demographic data, sensitization, Introduction: Pediatric asthma patients originating from poor- lung function, medication, peripheral eosinophilia, and markers of resource areas experience a significant increase of unscheduled and asthma severity. We performed hierarchical clustering (HC) using: (1) emergency department visits and most of medical care for them is all variables; and (2) variables transformed using dimensionality reduc- delivered in such a facility. Education and communication is vital for tion techniques. severe asthma patients. Virtual media can be used to improve asthma Results: PCA revealed 5 components describing atopy and variations control in these patients. Official data are documenting that ¾ of citi- in asthma severity, which were then used to infer cluster assignment. zens own at least a smart-phone in our country and even in poor areas The optimal HC solution in both PCA-transformed and raw untrans- there is excellent mobile phone coverage. The aim of our study was to formed data identified five clusters. However, these clusters were not document outcome of a structured tele-monitoring approach (via vir- identical. Both identified mild asthma with good lung function, severe tual media) in management of severe asthmatic children. atopic asthma and late-onset mild atopic asthma. However, the over- Method: Prospective 12  months study of patients with severe asthma. lap between children assigned to these three clusters in two HC analy- Were observed severe asthma patients from a cohort of previously mon- ses was modest. Clustering without PCA identified early-onset severe itored children with uncontrolled asthma [ACT child score below 19] atopic asthma and late-onset atopic asthma with high BMI, whilst and frequent visits in Emergency Department in previous year. Exten- early onset non-atopic mild asthma in females was identified in HC sive education in basic inhalation techniques [video-training] was done with PCA. and written-action plan (WAP) was provided. Were trained to use PIS Conclusion: Different methodologies applied to the same dataset [Pulmonary Index Score] to evaluate severity of exacerbation and treat- identified differing clusters of asthma. Despite cluster instability, both ment response. Parents had to either use ED or use WAP and tele-mon- methodologies provided meaningful clinical insights for understand- itoring. Short video of clinical status was evaluated by an expert and ing asthma heterogeneity. further options were discussed. Non-responders were presented in ED. Clin Transl Allergy 2017, 7(Suppl 2):14 Page 9 of 17 POSTER DISCUSSION SESSION 1—Biomarkers generated sequences and/or number of samples. A correlation matrix was used to identify positive and negative correlations between bac- terial and viral families and the results were superimposed in a net- P11 work graph. Th17 in children with asthma 1 1 1 Results: Taxon classification revealed 198 bacterial and 20 viral fami- Snezhina Lazova , Penka Perenovska , Guergana Petrova , Dimitrinka 1 2 3 3 lies. Unsupervised hierarchical clustering based on beta diversity (Jac- Miteva, Petar Velikov, Tsvetelina Velikova , Ekaterina Ivanova-Todorova , 3 3 card matrix) clustered controlled, partly controlled and uncontrolled Kalina Tumangelova-Yuzeir , Dobroslav Kyurkchiev 1 asthma separately. Topographical analysis of the metagenomic inter- Medical University, Pediatric clinic, UMHAT Alexandrovska, Sofia, Bulgaria; 2 3 action map identified the phage family of Myoviridae as a central Medical University, Sofia, Bulgaria; Medical University, Laboratory component. Further analysis of the Myoviridae family and its direct of Clinical Immunology, University Hospital St. Ivan Rilski, Sofia, Bulgaria interactions with bacterial families (17 families) revealed that patients Correspondence: Snezhina Lazova - snejina@lazova.com with partly controlled and uncontrolled asthma have reduced quantity Clinical and Translational Allergy 2017, 7(Suppl 2):P11 of Myoviridae phages and reduced diversity and quantity of their bac- terial interactors. Introduction: Th17 lymphocytes are now widely believed to be criti- Conclusion: Based on the Rarefaction curves, we estimate that, even cal for the regulation of various chronic immune diseases, including with 5 samples a high percentage of the microbial diversity can be asthma and chronic obstructive pulmonary disease. robustly represented. Asthma patients were efficiently clustered Objectives: To assess Th17 levels in children with asthma (BA) and based on their microbial similarity which seems to be representative cystic fibrosis (CF). of asthma control. The phage family Myoviridae is predicted to hold Method: We included 12 children with CF without history of allergies a central role in the viral-bacterial interaction on the specific environ- and 20 children with BA aged 7 to 16 years (10 with severe BA and 10 mental site. The reduced number of reads of the Myoviridae phages with moderate BA). After informed consent, 1.2  ml of venous blood along with the reduced diversity and quantity of 17 bacterial families was collected during a routinely performed blood withdrawal. Analy- in uncontrolled asthma could be evident of an ongoing ‘fight’ between ses of Th17 cells were performed with Lise-Wash Protocol by a 4-color components of the upper respiratory metagenome. FacsCalibur flow cytometer after staining the whole blood with the Keywords: Asthma, Metagenomics, Respiratory, Children following fluorescence-labeled antibodies: anti-CD3 (FITC), anti-CD (PerCP), anti-CD161 (PE) and anti-CCR6 (AlexaFluor 647). P13 Results: The patients in asthma group had significantly higher per - Circadian breathomics in asthma: analysis by thermal centage of Th17 12.40%  ±  1.16 compared to the children with CF - desorption‑gas chromatography‑mass spectrometry 7.64 ± 0.87 (p = 0.008). Stratifying the BA group according the severity 1 1 2 3 Maxim Wilkinson , Craig P ortsmouth , David Ray , Royston Goodacre , we found higher levels of Th17 in patients with severe BA (p  <  0.05). 4 2 Stephen J. Fowler , Hannah Durrington Patients with moderate asthma had TH17 values close to those in CF 1 Division of Infection, Immunity and Respiratory Medicine, Faculty of Biol- children. ogy, Medicine and Health, University of Manchester, Manchester, United Conclusion: The number of Th17 cells is significantly increased in the 2 Kingdom; Division of Metabolism and Gastroenterology, Faculty of Biol- peripheral blood of children with severe BA compared to the children ogy, Medicine and Health, University of Manchester, Manchester, United with moderate BA. Severe BA patients could have possible benefit 3 Kingdom; School of Chemistry, Manchester Institute of Biotechnology, from the future target therapies. 4 University of Manchester, Manchester, United Kingdom; Manchester Keywords: Severe Asthma, Controlled Asthma, Cystic Fibrosis Academic Health Science Centre, and NIHR Respiratory and Allergy Clinical Research Facility, University of Manchester, University Hospital of South Manchester, Manchester, United Kingdom Acknowledgements: This work was supported by a grant from the Correspondence: Maxim Wilkinson - maxim.wilkinson@postgrad. Medical University of Sofia (Council of Medical Science, project no. manchester.ac.uk 289/2015, grant no. 54/2015). Clinical and Translational Allergy 2017, 7(Suppl 2):P13 P12 Study of the viral and bacterial communities associated Introduction: Asthma is closely associated with the endogenous cir- with asthma: a metagenomic approach cadian rhythm of the lungs; in severe and uncontrolled asthma noctur- 1 1 Spyridon Megremis , Bede Constantinides , Alexandros Georgios nal wakening is common, as is a dip in early morning peak expiratory 1 2 1 Sotiropoulos , Paraskevi Xepapadaki , David Robertson , Nikolaos flow. This variability makes asthma a potential target for chronother - Papadopoulos apy to improve the efficacy of treatment. To personalise our approach 1 2 University of Manchester, Manchester, United Kingdom; University to chronotherapy in asthma, it is essential to have an easy-to-meas- of Athens, Athens, Greece ure biomarker that reflects an individual patient’s chronotype. This Correspondence: Spyridon Megremis - spyridon.megremis@manches- would allow chronotherapy to be targeted to the right time of day ter.ac.uk for each patient. Breath samples are easy to collect and non-invasive, Clinical and Translational Allergy 2017, 7(Suppl 2):P12 and exhaled volatile organic compounds (VOCs) have been related to airway inflammatory patterns in asthma. We hypothesise that the exhaled VOC profile contains potential biomarkers that track the cir - Introduction: An important unexplored source of interpersonal vari- cadian rhythm in asthma. Samples are analysed using thermal desorp- ation in asthma presentation is the complex and dynamic commu- tion-gas chromatography–mass spectrometry which is currently the nity of microorganisms located on different sites of the respiratory gold standard for offline VOC detection, and allows high sensitivity tract. Our aim was to identify and explore the bacterial and viral diver- and reliable identification of detected compounds. sity in a small pilot study of asthmatic children using metagenomics. Method: Twenty non-smoking volunteers (10 healthy, 10 with mod- Method: Nasopharyngeal samples were obtained from five asth- erate asthma) will be recruited. So far nine individuals have been matic children (2 males) at a baseline asymptomatic visit. Mean age recruited; the study design is detailed in the figure. Breath samples are was 5.2  years (range 4.6–5.8). Communities of viruses (DNA and RNA taken at 15:00 on the first visit; 15:00, 21:00, 03:00 and 09:00 on the genomes) and bacteria were evaluated using high depth paired- second and 09:00 on the final visit. This gives six samples per partici- end shotgun sequencing. We performed taxonomic classification of pant at four time points allowing investigation of both the changes in sequences and calculated environmental indexes (alpha and beta) to an individual’s VOC profiles over the course of a day and longitudinal estimate the microbial diversity both within (Shannon, Chao1, Jack1) variation over the course of several weeks. All samples are analysed and between (Jaccard) patients. Rarefaction curves were produced to using thermal desorption-gas chromatography-mass spectrometry study the relationship between microbial diversity and the number of Clin Transl Allergy 2017, 7(Suppl 2):14 Page 10 of 17 Results: The extracted ion chromatograms for the internal standard low dose ICS (n  =  11) therapy (p  =  0.011, p  =  0.007 respectively). (p-bromofluorobenzene), isoprene, limonene and the alkanes are Twenty AP (n  =  5, CA and n  =  15, UA) demonstrated impaired lung shown as a quality control procedure to ensure clean data process-function (FEV   <  80%), which was associated with higher levels of SP ing. Preliminary results will be presented by multivariate analysis for (p = 0.04). Similarly, in UA was observed significantly negative correla- the first nine participants comprising six individuals with asthma and tion between SP and FEV  % (p = 0.006, r = − 0.56). three controls. Conclusion: Patients with persistent moderate to severe bronchial Conclusion: Measuring exhaled breath VOCs at intervals through- asthma show higher levels of SP as compared to healthy controls. High out the day may provide a useful way of mapping a patient’s asthma levels of SP are associated with patient need of higher dose ICS ther- chronotype, guiding future chronotherapy. apy. Our results suggest that SP is biomarker for impaired lung func- Keywords: Circadian Rhythm, TD-GC–MS, Breathomics tion in poor asthma control. Keywords: Periostin, Severe Adult Asthma, Lung Function The research was funded by GRANT project No. 308/2015, contract No. 75/2015 to CMS, MU Sofia P15 Serum periostin values in healthy non‑asthma non‑copd spanish adults Irina Bobolea, Daiana Guillén Vera, Gabriel Gonzalez-Salazar, Carlos Melero Moreno, Consuelo Fernandez Rodriguez, Natividad De Las Cuevas Moreno Hospital 12 de octubre, i + 12 Research Institute, Madrid, Spain Correspondence: Irina Bobolea - ibobolea@gmail.com Clinical and Translational Allergy 2017, 7(Suppl 2):P15 Introduction: Serum periostin is an emerging biomarker for T2 asthma, and for the response to new biologicals such as anti IL-13 lebrikizumab. But a clear cut-off has not been established, and other pathologies beyond asthma are known to course with high periostin levels. The aim of this study was to establish reference values for serum periostin in a representative sample of non-asthmatic non-COPD Figure 4 Schematic of the experimental design healthy population in our geographical area. Method: A cross-sectional study was conducted between May–Octo- ber 2015. After signing the informed consent, we collected sera sam- ples from consecutive blood donors of the Blood Bank of Universitary Hospital 12 de Octubre, Madrid, Spain. By choosing this source of donors, all diseases and other conditions P14 related to any periostin elevations described so far were excluded, as Serum periostin levels as a biomarker for impaired lung function they constitute contraindications for donating blood: chronic illnesses in adult patients with moderate to severe asthma 1 1 2 (diabetes; heart, pulmonary, renal or hepatic diseases); any type of Denitsa Dimitrova, Vania Youroukova, Tsvetelina Velikova , Kalina 2 3 cancer; chronic or acute infectious diseases, including stomathology- Tumangelova-Yuzeir, Anna Valerieva 1 cal disordes; pregnancy; recent traumatic injury or surgery. Moreover, Medical Faculty of Medical University of Sofia; Clinical center of pulmo - 2 all participants completed a questionnaire aimed to exclude those nary diseases; SHATPD “St. Sofia”, Sofia, Bulgaria; Medical Faculty of Medi- with symptoms and/or a physician diagnosis of asthma and/or COPD. cal University of Sofia, Department of Clinical Laboratory and Clinical 3 Sera were frozen at −80 °C until periostin was measured using an EISA Immunology, University Hospital, Sofia, Bulgaria; Medical Faculty commercial kit. We calculated the sample size to estimate a media, of Medical University of Sofia, Clinic of Allergy and Asthma, University using a standard deviation (SD) of 15, with 3 ng/mL precision and 95% Hospital Alexandrovska, Sofia, Bulgaria confidence interval (Anastasilakis et al. 2014). Correspondence: Denitsa Dimitrova - ddimitrova87@abv.bg Results: 100 non-asthmatic non-COPD and otherwise healthy sub- Clinical and Translational Allergy 2017, 7(Suppl 2):P14 jects, aged between 18 and 65  years, were included in the study. Results are expressed in media (SD) (min–max range). Age: 43.97 years Introduction: The aim of the study is to determine the relationship (10.35) (21–65  years). 62 males (62%). 22 smokers (25.9%); being the between serum periostin (SP) levels and clinical characteristics in adult official percentage of smokers in the Madrid area 27.4%. patients with moderate to severe asthma (AP). Serum periostin: 33.04  ng/mL (14.37) (6.87–81.43  ng/mL). The media Method: SP levels were measured by ELISA method in 42 AP out of estimated in total population was (30.19–35.90  ng/mL) with a con- exacerbations and 10 healthy controls (HC). AP were divided into two fidence interval of 95%. Periostin in males: 33.51  ng/mL (14.02), and groups according asthma control test (ACT) score: controlled AP (CA, females: 32.28 ng/mL (15.09), p = 0.680. Smokers: 32.4 ng/mL (12.05) n = 19) and uncontrolled AP (UA, n = 23). Atopic status was confirmed and non-smokers: 33.09  ng/mL (15.28) respectively, p  =  0.850. No by skin prick testing and measurement of total and specific serum IgE association was either found between age and serum periostin levels. Lung functional parameters were assessed by spirometry. (r = 0.075). Results: We have observed significantly higher levels of SP in AP Conclusion: We report reference values for serum periostin in a (1.68  ±  0.52  ng/ml) compared to HC (1.08  ±  0.12  ng/ml) (p  <  0.001). healthy non-asthmatic non-COPD adult Spanish population. Our No statistically significant differences were found in SP levels between results could be used as reference for further studies aimed to clarify CA and UA, atopic (n = 14) and non-atopic AP (n = 28), current smok- the role of periostin in asthma phenotypes and endotypes, or to ers (n  =  13) and non-smokers (n  =  29) (p  =  0.85, p  =  0.33, p  =  0.19 choose future candidates for the new biological agents targeting T2 respectively). We have found significantly higher SP levels in AP on inflammation. medium (n  =  22) and high doses ICS (n  =  9) compared to those on Keywords: Asthma Biomarkers, Periostin Clin Transl Allergy 2017, 7(Suppl 2):14 Page 11 of 17 POSTER DISCUSSION SESSION 2—Mechanisms cells and is involved in lymphocyte activation, proliferation and differ - entiation. Targeting PI3  Kδ may have therapeutic benefit in currently uncontrolled asthma patients. P18 The aims of the study were to compare the expression PI3 Kδ in airway ToAST: Investigating the effects of bronchial thermoplasty (bt) samples from asthma patients and healthy subjects and to assess the on cough responses in patients with severe asthma – a pilot study anti-inflammatory potential of PI3  Kδ inhibitors on bronchoalveolar R. Wang, I. Satia, R. Niven, S. J. Fowler, D. M. Ryan, J. A. Smith lavage (BAL) lymphocytes. University Hospital of South Manchester, Manchester, United Kingdom Method: Bronchial biopsies, BAL cells and blood cells were collected Correspondence: Ran Wang - ranwang1986@googlemail.com from moderate to severe asthma patients and healthy subjects. The Clinical and Translational Allergy 2017, 7(Suppl 2):P18 numbers of PI3  Kδ-positive cells in biopsies was calculated using immunohistochemistry and the expression of PI3  Kδ in BAL CD3 cells Introduction: Severe asthma affects 5% of the asthma population but was assessed by flow cytometry. BAL and blood cells were treated with drives the majority of the morbidity and cost. Despite cough being a a PI3 Kδ inhibitor and/or dexamethasone prior to T cell receptor (TCR) major symptom and a marker of both severity and poor prognosis, stimulation. ELISAs were used to assess levels of IFNγ, IL-13 and IL-17 current medications are not designed to treat cough directly. Bron- and flow cytometry was used to measure pSTAT5, an early marker of chial thermoplasty (BT) is a novel therapy in severe asthma, deliver- T-cell activation. ing radiofrequency thermal energy to the large airways. There are 1 Results: Bronchial biopsies from asthma patients contained increased provisional data suggesting BT reduces the number of airway nerves . numbers of PI3 Kδ-positive cells compared to those from healthy sub- If this is the case, then BT may affect cough in asthma. Inhaled cap - jects, and asthma BAL T-cells expressed higher levels of PI3  Kδ than saicin challenge has been safely used to evoke cough responses and healthy cells. Inhibition of PI3  Kδ reduced TCR-stimulated IFNγ, IL-13 to assess cough threshold in research. However, to date no study has and IL-17 in BAL cells from both asthma patients and healthy subjects, adopted this method to assess cough in patients with severe asthma and treating BAL cells with a combination of PI3 Kδ inhibitor and dexa- or following BT. methasone had an additive effect on cytokine inhibition. TCR-induced We aim to assess the safety and feasibility of capsaicin challenge in pSTAT5 in purified T-cells and BAL cells was impeded by PI3 Kδ inhibi- people with severe asthma. We will also explore the effects of BT on tion and dexamethasone. 24 h cough frequency, capsaicin evoked cough responses and cough- Conclusion: Inhibition of PI3 Kδ in asthma may be an effective method related quality of life. of reducing activation of T-cells and subsequent cytokine production. Method: The protocol for this two-visit observational study has been Keywords: Phosphatidylinositol 3-Kinase Delta, Airway T-Cells, Bron- approved by the local Research Ethic Committee. All recruited patients choscopy, Corticosteroids, Cytokines (target n = 24, of whom half will have had BT ) will be treated at British Thoracic Society steps 4 or 5. During visit 1 patients will be consented, the Leicester Cough and Asthma Control Questionnaires administered P20 and baseline spirometry performed. They will undergo ambulatory Investigating neuronal responses by assessing capsaicin evoked 24-hour cough monitoring. A capsaicin challenge will be performed cough responses in an allergen challenge model of asthma (INCA) 1 1 1 2 during the second visit. Spirometry will be performed before and Imran Satia , Stephen Fowler, Mark Woodhead , Paul O’Byrne , Jaclyn immediately after the challenge to assess any change in FEV .Ann Smith 1 2 Results: The primary outcome measure will be the number of participants University of Manchester, Manchester, United Kingdom; McMaster with a 20% or greater fall in FEV1 and/or requiring rescue bronchodila- University, Hamilton, Canada tor therapy during the capsaicin challenge period. Secondary outcomes Correspondence: Imran Satia - imran.satia@manchester.ac.uk include: change in   %FEV1 predicted during capsaicin challenge; toler- Clinical and Translational Allergy 2017, 7(Suppl 2):P20 ability of inhaled capsaicin; number of adverse events during and after capsaicin evoked cough challenge; differences in capsaicin dose response Introduction: Cough is a common and troublesome symptom in curves between patients who have had BT versus those have not. asthma but little is known about the neuronal pathways that trigger Conclusion: This pilot study will lead to a larger prospective study cough. The mechanisms by which  airway inflammation, bronchial investigating the effects of BT on cough responses in patients with hyper-responsiveness and variable airflow obstruction cause cough severe asthma. is unclear. We have previously shown that methacholine induced bronchoconstriction heightens cough responses to inhaled capsaicin. Reference The aim of the current study was to investigate the effects of allergen 1. Bergqvist A, Pretolani M, Taille C, Dombret MC, Knapp D, Bjermer L, exposure on cough reflex sensitivity. Chanez P, Erjefalt JS, Aubier M. Selective structural changes of bronchial Method: We performed a 8 visit randomised, single-blind, placebo thermoplasty in the treatment of severe uncontrolled asthma. Am J controlled, two-way cross-over study comparing cough responses Respir Crit Care Med. 2015;191:A4171. to inhaled capsaicin in allergic asthma patients during and 24  h after exposure to allergen compared with diluent (saline) control. P19 Mild atopic asthmatics who are dual responders will undergo full dose Anti‑inflammatory capability of Phosphatidylinositol 3‑kinase allergen/diluent (saline) challenge. Early asthmatic responses (EAR) delta inhibitors in moderate to severe asthma patients (fall in FEV1 > 20% at 30 min) and late asthmatic responses (LAR) (fall 1 1 1 1 2 2 T. Southworth , J. Plumb, V. Gupta , J. P earson , I. Ramis , M. D. Lehner , in FEV1 > 15% at 3–7 h) will be documented. Full dose capsaicin chal- 2 1 M. Miralpeix , D. Singh lenge was performed at screening to determine the excitatory dose 1 2 The University of Manchester, Manchester, United Kingdom; Almirall S. evoking half the maximum cough response (ED50). The ED50 concen- A., Barcelona, Spain tration of capsaicin was inhaled 4 times, 30 s apart, at 30 min and 24 h Correspondence: Thomas Southworth - tsouthworth@meu.org.uk after inhaled allergen/diluent challenge. Cough counts will be meas- Clinical and Translational Allergy 2017, 7(Suppl 2):P19 ured throughout, along with methacholine challenge and induced sputum before and after allergen challenge. Introduction: Lymphocytes numbers are increased in the lungs of Results: We report interim analysis of the first 5 completed patients asthma patients, with T-helper 2 cells being associated with eosino- recruited to this study (mean age (SD) 23.4 yrs (4.6), BMI 23.9 (4.3), philic inflammation. There is also growing evidence that T-helper 1 FEV1% predicted 97.4% (8.0), PC20 1.55 (1.72), 2 female). All patients and T-helper 17 cells may play a role in more severe asthma. Inhaled were steroid naive and sensitive to house dust mites. Mean fall in corticosteroids (ICS) are widely used anti-inflammatory treatments for %FEV1 from baseline in the EAR was 34.9% (SD6.4), and LAR 29.2% asthma, but many moderate to severe asthma patients suffer from per - (20.6). Baseline ED50 concentration was 24.2microM(23.8). There was sistent symptoms, despite using high doses of ICS. Phosphatidylinosi- an increase in capsaicin evoked coughs after allergen exposure com- tol 3-kinase delta (PI3 Kδ) is predominantly expressed in inflammatory pared to diluent at both 30 min and 24 h (mean coughs after allergen Clin Transl Allergy 2017, 7(Suppl 2):14 Page 12 of 17 work is addressing the ability of Af activated DCs to influence T cell activation and polarization in vitro and in vivo. Keywords: Dendritic Cells, Bronchial Epithelial Cells, Aspergillus Fumigatus P22 Flavonoid cyanidin acts through IL‑17RA to alleviate IL‑17A‑mediated severe asthma Caini Liu, Liang Zhu, Kiochi Fukuda, Cunjin Zhang, Suidong Ouyang, Xing Chen, Luke Qin, Suguna Rachakonda, Mark Aronica, Jun Qin, Xiaoxia Li Figure 5 FEV1 and ED 50 Coughs 30 min and 24 h after Allergen/Diluent Cleveland Clinic, Cleveland, United States Correspondence: Caini Liu - liuc@ccf.org Clinical and Translational Allergy 2017, 7(Suppl 2):P22 vs. diluent at 30 min 15.0 (4.9) vs. 9.6(3.5), at 24 h 12.2 (8.0) vs. 6 (2.6). Introduction: Interleukin (IL)-17A plays a critical role in the pathogen- Figure 1 esis of asthma. High levels of IL-17A are found in bronchial biopsies Conclusion: We have safely performed capsaicin challenge after aller- and serum obtained from patients with severe asthma. Deficiency of gen exposure. Initial data suggests cough responses may be height- IL-17A signaling components attenuated airway inflammation in sev - ened for up to 24 h after allergen exposure. eral mouse models of asthma. We recently identified a potent small Our results show the importance of STAT6 and the possible implica- molecule compound A18 (called cyanidin, a key pigment present in tion of SOCS proteins in the enhanced CCL26 expression by BECs red berries and other fruits) for IL-17A inhibition through computer- from severe eosinophilic asthmatics (Supported by the Fondation de aided docking-based virtual screening. A18 specifically binds to a l’IUCPQ and the JD-Bégin Research Chair). region in the IL-17RA extracellular domain that overlaps with the Keywords: Asthma, Cough, Capsaicin, Allergen Challenge, TRPV1 binding site for IL-17A and this binding potently disrupts the IL-17A/ IL-17RA complex. We demonstrated that cyanidin effectively inhibited P21 IL-17A-mediated inflammatory gene expression in both human and Mechanisms of cross‑talk between pulmonary epithelial cells mouse cells. In the current study, we further investigated if A18 could and dendritic cells during type 2 inflammation attenuated airway inflammation in mouse models of steroid-resistant 1 1 1 1 Cecilia Forss , Peter Cook , Sheila Brown , Freya Svedberg , Katherine and severe asthma. 1 2 2 3 Stephenson , Margherita Bertuzzi , Elaine Bignell , Malin Enerbäck , Method: Adoptive transfer models for antigen‑induced airway 3 1 Danen Cunoosamy , Andrew M acdonald inflammation Manchester Collaborative Centre for Inflammation Research, Manches- OVA-specific Th17 and Th2 cells were transferred intravenously ter, United Kingdom; Manchester Fungal Infection Group, Manchester, (i.v.) into WT C57BL/6 female mice. Mice were challenged (i.n.) with United Kingdom; AstraZeneca R&D, Mölndal, Sweden OVA 1  day before transfer and 3 consecutive days after trans- 323–339 Correspondence: Cecilia Forss - cecilia.forss@postgrad.manchester.ac.uk fer. For A18 treatment, mice were injected (i.p.) with A18 on each Clinical and Translational Allergy 2017, 7(Suppl 2):P21 challenge day. Measurements were performed 24  h after the last challenge. Introduction: Everyday exposure to potentially harmful pathogens is High‑fat‑ diet (HFD) intervention carefully monitored and controlled by the interaction between a num- Starting at 4  weeks of age, WT C57BL/6 male mice were fed with ber of different cell types, such as epithelial cells and dendritic cells 10  kcal   % fat chow diet (CD) or a 60  kcal   % HFD for 14  weeks. A18 (DCs). Both have the ability to sense environmental changes, and initi- group of mice were injected (i.p.) with A18 daily for 4  weeks before ate and direct an immune response. This project is focusing on charac- subjection to measurements. terizing the interactions between epithelial cells and DCs in the early House dust mite (HDM)‑induced asthma stages of an allergen exposure. Aspergillus fumigatus (Af) is an oppor- WT C57BL/6 female mice were sensitized (s.c.) with HDM (100  µg/ tunistic fungus with the capacity to cause a range of responses, from mouse) in Complete Freund’s Adjuvant on day 0 and subsequently low-level inflammation in healthy people to life-threatening condi- challenged (i.n.) with HDM (100  µg/mouse) on day 14. A18 group of tions in the immunocompromised. mice were injected (i.p.) with A18 on days 13 and 14 and also adminis- Method: We have concentrated on understanding early events in the trated (i.n.) with A18 1 h before challenging. Measurements were per- immune response towards Af, with a current emphasis on DCs. formed 24 h after the last challenge. Results: 4 h post exposure, ~ 50% of in vitro generated DCs were posi- Results: tive for Af spores and the majority of the spores were internalised. Af A18 inhibited Th17-induced but not Th2-induced airway inflammation. positive DCs showed higher expression of CD40, CD80 and CD86 com- A18 substantially attenuated steroid-resistant obesity-associated pared to cells that were spore negative. Interestingly, while Af induced asthma. intermediate up-regulation of DC activation markers, it induced only A18 alleviated House dust mite (HDM)-induced neutrophilic airway low levels of cytokine production. As the type of interaction between inflammation. DCs and Af likely influences downstream activation and polarization of Conclusion: The findings strongly suggest that A18 (cyanidin) can be T cells, we are currently dissecting the impact of internalised vs. sur- used as the prototype for developing small molecule drugs for treat- face bound spores, compared to soluble fungal extracts, on activation ing the IL-17A-mediated severe asthma. of DCs in  vitro. In parallel, we have used murine GM-CSF- and Flt3-L- Keywords: Cyanidin, IL-17A, Severe Asthma derived DCs and human monocyte derived DCs, as well as human bronchial epithelial cells (HBECs) in air–liquid interface (ALI) cultures. P23 Additionally, we have used a murine model of intranasal exposure to Mechanisms involvement in CCL26 production by bronchial low doses of live Af to investigate Af uptake by, and activation of, pul- epithelial cells: importance in asthma and its severity monary DC subsets and epithelial cells in vivo. Marie-Chantal Larose, Anne-Sophie Archambault, Véronique Provost, Conclusion: Data generated so far indicate a fungal life stage-depend- Jamila Chakir, Michel Laviolette, Nicolas Flamand ent up-regulation of activation markers, accompanied by induction CRIUCPQ, Québec, Canada of low-level secretion of inflammatory cytokines. Af exposure in  vivo Correspondence: Marie-Chantal Larose - marie-chantal.larose@criucpq. resulted in marked airway inflammation with infiltration of immune ulaval.ca cells, goblet hyperplasia and increased collagen deposition. Ongoing Clinical and Translational Allergy 2017, 7(Suppl 2):P23 Clin Transl Allergy 2017, 7(Suppl 2):14 Page 13 of 17 Introduction: High pulmonary eosinophil counts correlate with Results: In this lung infection model, anti-Ym1 treatment led to a asthma severity and exacerbation. We recently showed that sputum reduction in IL-17A expression and a reduction in the number of CCL26 levels correlate with sputum eosinophils. We also found that innate  γδ T cells during the early stages of infection. The altered IL- among all CC chemokines, IL-13 selectively induced the expression of 17A response resulted in decreased IL-13 and IL-5 mRNA expression in CCL26 by bronchial epithelial cells (BECs) and this phenomenon is sig- whole lung tissue and reduced numbers of IL-5 + and IL-13 + CD4 + T nificantly enhanced in BECs from severe eosinophilic asthmatics. We cells and innate lymphoid cells; results mimicked by global IL-17A postulated that the superior CCL26 production that we observed in deficiency. severe asthmatics was the consequence of increased signaling events Conclusion: CLPs impaired the development of type 2 response by mediated by IL-13. We thus assessed the expression and functional modulating IL-17 expression. This suggests that CLPs can regulate responses of the different signaling effectors linked to the IL-13 signal- both innate and adaptive immune mechanisms that may be key in ing in CCL26 expression by BECs from healthy subjects, mild asthmat- tipping the balance of airway inflammation towards neutrophil domi- ics, and severe eosinophilic asthmatics. nance during allergic lung pathology. Method: Human primary BECs were isolated and cultured from bron- Keywords: Neutrophil, IL-17, Chitinase-Like Protein, Lung chial biopsies. BECs were treated with IL-13 or vehicle for different Inflammation times. Inhibitors or their vehicles were added to BECs 1 h before IL-13. References: CCL26 expression was assessed by qPCR and ELISA. STAT6 and phos- 1. Chupp GL et al. N Engl J Med. 2007;357:2016–27 phorylated (p)STAT6 were quantitated by ELISA in BEC lysates. SOCS3 2. Sutherland TE et al. Nat Immunol. 2014;15:1116–25 level were assessed by immunoblotting. Results: We confirmed the involvement of STAT6 in the induction of CCL26 expression by treating BECs with increasing STAT6 inhibitor. POSTER DISCUSSION SESSION 2—Treatment This led to a concentration-dependent inhibition of CCL26 expression in IL-13-stimulated BECs. In that regard, the pSTAT6/STAT6 ratios were P25 increased in IL-13-stimulated BECs from severe eosinophilic asthmat- Once‑daily tiotropium respimat add‑on therapy improves lung ics, compared to those from healthy subjects and mild asthmatics. This function in patients aged 6–17 years with severe symptomatic increased activation of STAT6 might be explained by decreased SOCS3 asthma level in BECs from severe eosinophilic asthmatics. Also, we confirmed 1 2 3 4 4 E. Hamelmann, C. Vogelberg , S. Goldstein , G. E. Azzi , M. Engel , R. the importance of STAT6 and IL-13 in CCL26 expression and produc- 5 6 Sigmund , S. J. Sz efler tion in IL-13-treated-BECs throughout the incubation time. 1 Children’s Center, Evangelisches Krankenhaus Bielefeld, and Allergy Keywords: CCL26, Eosinophils, Bronchial Epithelial Cells 2 Center of the Ruhr University, Bochum, Germany; University Hospital Carl Gustav Carus, Technical University of Dresden, Dresden, Germany; Island P24 Medical Research, Rockville Centre, New York, New York City, New York, Chitinase‑like proteins: the missing link in allergen‑induced United States; Boehringer Ingelheim Pharma GmbH & Co. KG, Ingelheim neutrophilic inflammation? Am Rhein, Germany; Boehringer Ingelheim Pharma GmbH & Co. KG, 1 2 2 3 6 Nicola Logan , Dominik Ruckerl , Judith E. Allen , Tara E. Sutherland Biberach An Der Riss, Germany; Children’s Hospital of Colorado and the Institute of Immunology and Infection Research, University of Edin- University of Colorado School of Medicine, Aurora city, Colorado, United burgh, Edinburgh, United Kingdom; Faculty of Biology, Medicine States & Health, University of Manchester, Manchester, United Kingdom; Correspondence: Eckard Hamelmann - eckard.hamelmann@evkb.de Manchester Collaborative Centre for Inflammation Research, University Clinical and Translational Allergy 2017, 7(Suppl 2):P25 of Manchester, Manchester, United Kingdom Correspondence: Tara E. Sutherland - tara.sutherland@manchester.ac.uk Introduction: Tiotropium Respimat (tioR) add-on therapy to inhaled Clinical and Translational Allergy 2017, 7(Suppl 2):P24 corticosteroids (ICS) with or without additional controllers has been shown to improve lung function in Phase II and III studies of adults, adolescents and children with symptomatic asthma. We present a Introduction: Immune responses during asthma are typically consid- pooled analysis of lung function data in adolescents and children with ered as T-helper 2 (Th2) type responses marked by increased eosino- severe symptomatic asthma. phils. However, inflammation in severe asthmatics is often defined by Method: Two Phase III, randomised, double-blind, placebo-controlled, neutrophil or mixed neutrophil/eosinophil airway inflammation, along parallel-group, 12-week trials in patients aged 6–11  years (Viva- with increased IL-17 cytokine levels and airway remodelling. Little is TinA-asthma; NCT01634152) and 12–17  years (PensieTinA-asthma; known about the triggers that make neutrophils/IL-17 dominant char- NCT01277523) with severe symptomatic asthma. Patients received acteristics in severe asthmatics, or how these responses contribute to once-daily tioR 5 µg (2 × 2.5 µg), tioR 2.5 µg (2 × 1.25 µg) or placebo pathology, especially in scenarios when Th2-inflammation remains Respimat (pboR) as add-on to high-dose ICS plus another control- prevalent. Chitinase-like proteins (CLPs) are established markers of ler or as add-on to medium-dose ICS plus two other controllers. ICS immune activation and pathology. Of note, whilst considered Th2- dose was as defined in GINA 2009 (PensieTinA) or 2010 (VivaTinA) driven molecules, CLPs are also associated with IL-17 and neutrophilic guidelines. Patients were required to have a  ≥  3-month (PensieTinA) inflammation. In asthmatic patients, enhanced levels of circulating or  ≥  6-month (VivaTinA) history of asthma and be symptomatic at CLP YKL-40 correlates with disease severity and airway neutrophilia . screening and before randomisation by Asthma Control Questionnaire In addition, we recently described a novel role for murine CLPs in (interviewer-administered; VivaTinA) mean score  ≥  1.5. Primary end driving alveolar neutrophil recruitment through increased IL-1/IL-18 point of both studies: change from baseline (response) in peak forced expression and expansion of IL-17A-producing γδ T cells . Consider- expiratory volume in 1  s within 3  h post-dose (peak FEV ); key 1(0–3h) ing their association with both Th2 pathology and IL-17A/neutrophils, secondary end point: trough FEV response (measured 10 min before we sought to examine whether CLPs might link IL-17 responses to Th2 next dose of study medication); post hoc end point: trough FEV / inflammation. forced vital capacity (FVC) ratio, all measured at Week 12. Method: We explored the effects of neutralising the most abun- Results: 793 participants (VivaTinA: 401; PensieTinA: 392) were ran- dant murine CLP Ym1, with monoclonal antibody treatment during domised across both trials; 792 were included in this pooled full infection with the rodent lung migrating nematode N. brasiliensis. analysis set. Baseline demographics and disease characteristics were Infection with N. brasiliensis  in mice elicits lung immune responses balanced between treatment groups. TioR add-on therapy improved similar to that observed in allergic airway inflammation, thereby mak - lung function in the pooled population at Week 12, with tioR 5  µg ing it a valid model to study the interaction between IL-17A and Th2 showing superior improvements in peak FE V response, trough 1(0–3h) inflammation. Clin Transl Allergy 2017, 7(Suppl 2):14 Page 14 of 17 Table 3 Treatment results Response (change Peak Trough FEV Trough FEV / 1 1 from baseline) FEV response (mL) FVC ratio (%) 1(0–3h) measured response at Week 12 (mL) Tiotropium Respimat 117 ± 34 71 ± 34 1.921 ± 0.666 5 µg once daily (51, 183) (3, 139) (0.614, 3.229) (n = 260), adjusted p = 0.0005 p = 0.0395 p = 0.0040 mean of difference versus placebo Respimat ± SE Figure 6 Patient 1 and patient 2 (95% CI) Tiotropium Respimat 74 ± 33 64 ± 34 1.930 ± 0.666 2.5 µg once daily (8, 140) (−3, 132) (0.623, 3.236) (n = 263), adjusted p = 0.0273 p = 0.0617 p = 0.0038 ventilation, IMV was initiated. On Day-1, despite medical therapy and mean of difference IMV optimizations, severe dynamic hyperinflation with refractory versus placebo hypoxemia persisted and VV-ECMO was initiated. Thoracic CT revealed Respimat ± SE almost complete bilateral lung collapse. Methicillin-sensitive S. aureus (95% CI) and S. pneumoniae were isolated in endotracheal aspirate. Normal Full analysis set. CI, confidence interval; SE, standard error lung function was progressively reestablished following considerable viscid mucus secretions elimination and antibiotherapy. ECMO-VV was discontinued on Day-18. Extubation, ICU and hospital discharges occurred on Day-24, -27 and -55, respectively. FEV response and FEV /FVC ratio versus pboR, and tioR 2.5 µg show- 1 1 Conclusion: How this report contributes to current knowledge. Dur- ing superior improvements in peak FE V response and FEV / 1(0–3h) 1 ing refractory NFA, low-flow ECCO R allows correction of severe res- FVC ratio versus pboR, with numerical improvements in trough FEV piratory acidosis and facilitates extubation while high-flow VV-ECMO response versus pboR (Table). Safety and tolerability of tioR in both tri- is needed when significant atelectasis complicates airway narrowing als was comparable to placebo. and severe hypoxemia ensues. Knowledge of respiratory failure patho- Conclusion: Tiotropium Respimat add-on therapy is an effective physiology in refractory NFA allows the correct use of different ECLS bronchodilator, producing clinically meaningful improvements ver- modalities as a bridge to recovery. sus placebo in lung function in patients aged 6–17  years with severe symptomatic asthma, mirroring findings in adult patients with symp - tomatic asthma. Consent to publish: Consent to publish was received from the Keywords: Asthma Management, Bronchodilators, Children patients. P26 P27 Near‑fatal asthma: differential role of ECCO2R and VV‑ECMO Clinical and functional characteristis of 4 severe asthmatic as rescue therapies patients with absence of response to omalizumab 1 2 2 2 Raquel Mesquita , Luis Coentrão, Rui Veiga , José-Artur Paiva , Roberto Wendy Vargas Porras, Ana González Moreno, Jesus Macías Iglesias, Roncon-Albuquerque Jr Gustavo Córdova Ramos, Yesenia Peña Acevedo, Miguel Angel Tejedor Internal Medicine Department, Centro Hospitalar de São João, Porto, Alonso, Maria Del Mar Moro Moro Portugal; Emergency and Intensive Care Medicine Department, Centro University Hospital Foundation Alcorcón, Madrid, Spain Hospitalar de São João, Porto, Portugal Correspondence: Wendy Vargas Porras - wendy.drawenwen@gmail.com Correspondence: Raquel Mesquita - raquelmsmesquita@gmail.com Clinical and Translational Allergy 2017, 7(Suppl 2):P27 Clinical and Translational Allergy 2017, 7(Suppl 2):P26 Introduction: 69.9% of patients with severe asthma respond to Introduction: During a near-fatal attack of asthma (NFA) alveolar omalizumab, when it is used according to International Guidelines of hypoventilation underlies respiratory acidosis while severe hypoxemia asthma. indicates intrapulmonary shunt. Illustrate the differential role of low- We describe clinical and functional characteristics of 4 patients with flow extracorporeal CO removal (EC CO R) and high-flow veno-venous severe asthma treated with omalizumab and belong to our series of 2 2 extracorporeal membrane oxygenation (VV-ECMO) as rescue therapies severe asthmatic patients treated with this drug. in NFA. Method: We collected 24 patients with severe asthma treated with Method: Patient 1 A 31-year-old male with early-onset asthma and omalizumab from the Allergy Unit of the University Hospital Founda- epilepsy was admitted with acute respiratory failure (ARF) compli- tion Alcorcón, during the period 2007 to 2015. Of these, 4 patients did cated by generalized tonic–clonic seizure. He presented severe acute not respond to treatment. The “No responders”  was defined by the respiratory acidosis without hypoxemia after intubation (pH-7.10 responsible doctor, based on the clinical and functional assessment of pCO -82  mmHg P/F 556). Thoracic CT: diffuse bronchial wall thick - each patient considering several clinical and functional characteristics: ness without atelectasis. On Day-5 dynamic hyperinflation (auto- beta-adrenergic use, number of exacerbations and scores ACQ and PEEP 10 cmH O) under deep sedation and neuromuscular blockade ACT, other comorbidities, spirometric and lung volumes. These values persisted, precluding weaning from invasive mechanical ventilation were measured in the last year before the start of omalizumab and the (IMV). ECCO R was initiated. Extubation, active physical therapy and final year of treatment with omalizumab. full patient mobilization were possible on Day-6. EC CO R discon- Results: In 4 patients it was withdrawn omalizumab because of no tinuation, ICU and hospital discharges occurred on Day-8, -9 and -11, efficacy. Responders and non-responders were treated, at least, during respectively. 1  year. The non-responders had more non-respiratory comorbidities Results: Patient 2 A 34-year-old obese female with late-onset and than responders (75%-25%,p  =  0.07). However responders, seemed poorly controlled asthma was admitted with ARF with severe hypox- to have more severe asthma, with a greater number of exacerbations emia (pH-7.45 pCO -30  mmHg P/F 69). After a trial of non-invasive (responders 3.24 vs no responders 2.75), more use of beta-adrenergic 2 Clin Transl Allergy 2017, 7(Suppl 2):14 Page 15 of 17 (responders 2.47 vs no responders 1), more cycles oral corticoster- However, determination of inflammation using patient-based symp - oids (responders 2.64 vs no responders 1.5) and higher ACQ score tom assessment or spirometry is difficult and frequently underesti- (responders 2.1 vs no responders 1.8). Only use of oral corticosteroids mates asthma severity. Measurement of exhaled nitric oxide (FeNO) and use of beta-adrenergic the differences were significant (p  =  0.02 can support the clinical assessment of the patient at the point of and 0.05 respectively). However the residual volume  (RV) higher in care and can provide insights into underlying airway inflammation non-responders (responders 188% vs no responders 154%), although and therefore potentially help practitioners optimize their treatment differences were not statistically significant (p = 0.3). decisions. High FeNO concentrations have been associated with more In the last year of use of omalizumab, asthma was worse in the group severe/uncontrolled asthma and poor compliance with ICS therapy. of non-responders, with increased use of beta-adrenergic (p =  0.005) Method: The objective of the ASTH MA survey was to explore the real and greater use of steroids cycles (p = 0.002). world impact of measuring FeNO on physicians’ treatment decisions. Conclusion: In non-responders patients, the RV was higher and also Physicians initially recorded their assessment of airway inflammation had more comorbidities. (low, intermediate or high) based on the patient’s clinical presentation The small number of our sample limits the validity of our findings. and then by measuring patient’s FeNO using an approved electro- chemical device. Based on the FeNO result, physicians recorded what changes in drug therapy were made. P28 Results: Data from 337 physician practices, which included 7,901 One year follow‑up of patients after omalizumab treatment patients with asthma, were available for analysis. Assessment of withdrawal inflammation using clinical impression vs FeNO measurement was Irena Krcmova, Jakub Novosad determined to be low in 4,247 patients (53.8%) vs 5,083 (64.3%) Institute of Clinical Immunology and Allergy, University Hospital, Hradec (< 25 ppb); intermediate in 2,749 (34.8%) vs 1,802 (22.8%) (25–50 ppb) Kralove, Czech Republic and high in 905 (11.5%) vs 1016 (12.9%) (> 50 ppb). Mean high FeNO Correspondence: Irena Krcmova - irena.krcmova@fnhk.cz was 87  ppb (range 51–300, median 77) with FeNO  ≥  51–99  ppb in Clinical and Translational Allergy 2017, 7(Suppl 2):P28 n  =  760, FeNO 100–199  ppb in n  =  229 and FeNO 200–300  ppb in n = 27 patients. Clinical impression matched actual FeNO of < 25 ppb Introduction: Omalizumab has demonstrated its clinical efficacy in in 64.4%, 25–50  ppb in 46.9% and  >  50  ppb in only 33.6%. Changes asthmatic patients in number of studies. However, data about its clini- in treatment were made in 68.4% (695/1016) of patients who had a cal benefits after treatment cessation are still missing. To stress this high FeNO (>  50  ppb); stepping up anti-inflammatory treatment in challenging clinical issue, we have arranged a pilot observational- 96.1% and stepping down in 2.9%. Inhaled steroids were started in 345 study following clinical and laboratory data of 12 patients with severe patients and increased in 213 patients. Oral steroids were increased/ uncontrolled atopic asthma treated with omalizumab (11–61 months) started in 110 patients. Omalizumab was started in 2 patients. for one year after treatment withdrawal due to clinical stability. Conclusion: Our results suggest that measurement of FeNO at point Method: Study had a mixed design with repeated measures at 4 time of care improves the clinical assessment of underlying airway inflam- points, at treatment initiation, directly after treatment cessation, and mation in asthma and leads to clinically relevant changes in treatment 6 and 12  months thereafter. We have focused on clinical (pulmonary especially in those patients presenting with high FeNO (> 50 ppb). functions, inhalation corticosteroid (ICS) doses, asthma control test Keywords: FeNO, Asthma, Management, ASTH2MA Survey (ACT ), skin prick tests positivity) and laboratory (FeNO, eosinophils and total IgE levels) parameters. Statistical analysis of collected data has P30 been undertaken. Repeated measures were treated using a general 4‑month omalizumab efficacy outcomes for inadequate linear model or Friedman ANOVA in case of normality assumption vio- controlled severe allergic asthma in the Netherlands 2012–2015 lation. Post-hoc analysis was applied using Wilcoxon signed-rank test Sanne Snelder, Gert-Jan Braunstahl with Bonferroni adjustment. Sint Franciscus Gasthuis, Rotterdam, The Netherlands Results: We have demonstrated a significant reduction of ICS doses Correspondence: Sanne Snelder - sannesnelder@hotmail.com and SPT positivity (wheal diameter/mm) and increase of ACT dur- Clinical and Translational Allergy 2017, 7(Suppl 2):P30 ing omalizumab treatment regardless of a dominant allergic linkage, treatment duration or omalizumab dose. This effect has been visible even 6  months after treatment withdrawal, but only ACT sustained Introduction: Since 2006, omalizumab has been prescribed for inad- increased significantly 12  months after treatment cessation. Two equately controlled severe allergic asthma in the Netherlands. In 2011, patients had a treatment of oral corticosteroids, which were perma- more stringent rules were applied by Dutch healthcare policy makers. nently tapered during the omalizumab therapy and subsequently. This implied that omalizumab should be strictly prescribed within the Conclusion: We conclude, that omalizumab treatment outcomes registered label in exchange for reimbursement: inclusion criteria were reach clinical and statistical significance even one year after treatment a positive skin test or in  vitro activity to a perennial aeroallergen, a withdrawal. “Step down” of omalizumab in “treatment responders” FEV1 less than 80 percent, more than 2 severe exacerbations and sub- remains a highly individual process that can undergo by mutual agree- stantial symptoms despite treatment with inhaled corticosteroids (ICS) ment with the patient under clearly defined treatment plan. and long-acting B2-agonists (LABAs). Keywords: Asthma, Omalizumab, Withdrawal, Clinical Effect To evaluate the 4-month efficacy outcomes of omalizumab treatment for inadequate controlled severe allergic asthma after the stringent rules were applied by the Dutch healthcare policy. P29 Method: This is a “real world” prospectively designed data registry High FeNO leads to changes in asthma treatment: cohort analysis in which the outcomes of patients (2012–2015) who received omali- from the assessment of TH2 inflammation (ASTH2MA) survey 1 2 2 2 zumab were evaluated after the policy change. The primary endpoint Nicola Alexander Hanania , Marc Massanari , Heike Hecker , Eric Kassel , 3 4 was a good or excellent response to omalizumab after 16  weeks as Craig Laforce , Kathy Rickard 1 2 scored by the treating physician. Secondary endpoints include change Baylor College of Medicine, Houston, Texas, United States; Circassia, 3 from baseline to week 16 in the Asthma Control Questionnaire (ACQ), Raleigh, North Carolina, United States; North Carolina Clinical Research, 4 FEV1 scores and oral corticosteroids (OCS) use. Raleigh, North Carolina, United States; Circassia, Raleigh, North Carolina, Results: 403 patients had a complete data set and could be evalu- United States ated. 63% percent of the patients had a good or excellent response Correspondence: Heike Hecker - heike.hecker@circassia.com to omalizumab after 16 weeks. 85.5% of the responders showed more Clinical and Translational Allergy 2017, 7(Suppl 2):P29 than 0.5 points improvement in the ACQ score at 16 weeks. The mean FEV increased from 71.58 to 79.06. 63% of the responders had an Introduction: Recognizing TH inflammation in asthma has led to improvement of 5% of the FEV1. The maintenance OCS use was lower improvements in optimization of drug therapy and disease control. at 16  weeks. There was no relation between FEV1  <  80 and ≥  80% at Clin Transl Allergy 2017, 7(Suppl 2):14 Page 16 of 17 Table 4 Systematic review Trial Kuehr Casale Kopp 2008 Massanari et al 2006 2010 Disease Seasonal Seasonal Seasonal rhinitis persistent rhinitis rhinitis and asthma asthma Severity Moder- Moderate mild asthma At least ate- moderate severe Age range 6–17 18–50 11–46 18–55 Total 221 159 140 275 number of patients Site of study Germany USA Germany USA Duration of 36 weeks 22 weeks 20 weeks 26 weeks enrolment Figure 7 Improvement of ACQ and FEV1 compared to response Allergen Grass and aqueous Depigoid,modified Cat, dog used birch short grass pollen and HDM extract rag- extract extract Table 3 Response vs FEV1 < 80 weed extract Response Protocol of Conven- Rush Pre-seasonal rush Cluster immuno- tional protocol titration protocol immuno- YES NO Total therapy therapy FEV1 < 80 YES 178 80 258 Length of 14 weeks 10 weeks 8 weeks Perennial pre- allergen FEV1 < 80 NO 73 33 106 seasonal Total 251 113 364 immuno- P = 0.981 therapy Time of 12 week Pre-treat- Co-administration Pre-treat- adminis- after ment ment for Table 4 Response vs year of inclusion tration of immu- 9 weeks 13 weeks Omali- nother- Year of inclusion Response No response zumab apy started 2012 (%) 63.6 36.4 Dose of 0.016 mg/ 0.016 mg/ Dose/kg/IgE IU/mL 0.016 mg/ 2013 (%) 70.7 29.3 Omali- kg per kg/IgE every 2–4 weeks kg/IgE 2014 (%) 64.9 35.1 zumab IU/Ml IU/mL IU/mL every 2 or 2015 (%) 69.2 30.8 4 weeks P = 0.690 Introduction: Oral corticosteroids (OCS) are used in the management baseline and response after 4  months (Table  3). The response rate of both acute and severe asthma. Acute courses are typically well toler- remained stable over the years 2012–2015 (Table  4). Figure  1 shows ated, but well documented side effects occur in prolonged treatment. that most of the responders had an improvement of both FEV1 and Side effects include skin thinning, cataracts, diabetes, osteopenia and ACQ. It also shows that ACQ appears to be a better measurement for a easy bruising; these may impact adherence to treatment. Patients response than FEV1. rarely admit poor adherence to treatment but, with increasing avail- Conclusion: 63% of the 403 patients with inadequately controlled ability of monoclonal antibody therapy, adherence should be good severe allergic asthma had a good or excellent response to omali- prior to escalation to high-cost treatment. Mass-spectrometry can zumab after 16  weeks. Overall the ACQ improved, FEV1 increased assess concentrations of cortisol and prednisolone in peripheral blood, and there was lower use of OCS at 16  weeks. There was no relation- and this can provide evidence of OCS adherence as well as assessing ship between patients with a FEV1 < 80 and ≥ 80% at baseline and the absorption. We conducted an audit of results of all our prednisolone response rate. Improvement of ACQ appears to be a better measure- assays conducted in patients with severe asthma. ment for response than improvement of FEV1. Method: Data was collected on all patients with severe asthma under- Keywords: Asthma, Omalizumab going prednisolone assays at our centre. OCS assays were performed prior to escalation of therapy where there was a suspicion of poor P31 adherence. The results of the assay, participant age, gender, BMI, FEV1, Adherence to oral corticosteroids; audit results of prednisolone Fractional Exhaled Nitric Oxide (FeNO), eosinophil count and predniso- assays lone dose were recorded. These were used to determine which patient T. L. Jones, D. Neville, E. R. Heiden, E. Lanning, T. Brown, H. Rupani, K. S. characteristics correlate with adherence. Babu, A. J. Chauhan Results: 14 adults taking oral prednisolone were included in this Portsmouth Hospitals NHS Trust, Portsmouth, United Kingdom audit. This comprised 11 females and 3 males with a mean age of Correspondence: Thomas Llewelyn Jones - thomas.jones@porthosp.nhs.uk 46.3  ±  13.4  years, a mean prednisolone dose of 20.2  ±  10.4  mg, a Clinical and Translational Allergy 2017, 7(Suppl 2):P31 Clin Transl Allergy 2017, 7(Suppl 2):14 Page 17 of 17 mean BMI of 30.7  ±  7.2, a mean FeNO of 79.9  ±  53.2, mean FEV1 of The purpose of this review is to establish whether the combination of 64 ±  17.6% predicted and a mean eosinophil count of 0.35 ±  0.34. 5 omalizumab and subcutaneous allergen immunotherapy (SCIT ) has an out of 14 were compliant as assessed by a supressed cortisol (5/5) or add-on benefit to the efficacy and safety of SCIT in asthma and allergic detectable blood prednisolone prior to dosing (4/5). All 14 patients rhinitis? absorbed prednisolone well following oral OCS administration. Factors Method: This systematic review included double-blind randomized correlating with compliance included gender (males 66% compliant, placebo controlled trials of the add-on benefits of the combination of females 27% compliant), BMI (25.5 kg/m2 compliant vs 33.5, p = 0.05) omalizumab and SCIT compared to SCIT alone. and peripheral eosinophil count (compliant 0.15 vs 0.45, p = 0.09). Results: The combination of omalizumab and SCIT has decreased Conclusion: Only 36% of patients in our audit were adherent to OCS symptoms score, rescue medication score, and symptoms load in highlighting the importance of checking medication adherence dur- patients with rhinitis and patients with rhinitis and co-existing asthma. ing each clinical review, particularly when considering patients for In patients with asthma and those with rhinitis treated with rush novel therapies. Male gender, lower BMI and low eosinophil count immunotherapy, the percentage of patients developed systemic reac- were associated with compliance, but FeNO, FEV1 and OCS dose were tion to SCIIT including severe reaction requiring adrenaline admin- not. We suggest an objective assessment of compliance prior to initia- istration was significantly decreased in the combination treatment tion of expensive novel biological agents. group compared with SCIT/placebo group. Omalizumab has reduced Keywords: Prednisolone, Medication, Adherence, Education the risk for anaphylaxis by 5 folds in rhinitis patients treated with rush immunotherapy protocol. Only one trial reported the of number of adrenaline doses used during the trial P32 Conclusion: The addition of omalizumab has increased the efficacy of Does the combination of omalizumab and Subcutaneous allergen SCIT in patients with rhinitis and patients with rhinitis and co-existing immunotherapy (SCIT) have an add‑on benefit to the efficacy asthma. Omalizumab also reduced the risk of serious systemic reaction and safety of allergen immunotherapy in asthma and allergic in patients with asthma and in rhinitis patients treated with rapid high rhinitis? systematic review 1 2 3 2 dose of allergen. Addition of omalizumab has helped more patients at M. Y. Eldegeir , A. A. Chapman , M. Ferwana , M. Caldron 1 2 high risk of adverse effects related to SCIT to reach their target immu- National Guard Hospital, Dammam, Saudi Arabia; Imperial Collage 3 notherapy. An important findings of this review is that the use of London, London, United Kingdom; King Saud University, Riyadh, Saudi adrenaline is under-reported by immunotherapy clinical trials Arabia Keywords: Omalizumab, Immunotherapy, Asthma, Rhinitis, Allergic Correspondence: Manal Yousif Eldegeir - almanjil@gmail.com Clinical and Translational Allergy 2017, 7(Suppl 2):P32 Publisher’s Note Introduction: The last two decades witnessed a substantial expansion Springer Nature remains neutral with regard to jurisdictional claims in pub- in our understanding of the pathoimmunological mechanism of aller- lished maps and institutional affiliations. gic respiratory diseases. This led to a huge surge of interest in devel- oping novel treatment modalities aiming for improved efficacy and safety of AIT. Co-administration of anti IgE is an attractive approach with the added benefit of omalizumab as a biological agent which has its own independent Immunomodulation effect on asthma and rhinitis.

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Published: May 24, 2017

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