Abstracts from the 11th Symposium on Experimental Rhinology and Immunology of the Nose (SERIN 2017)

s from the 11th Symposium on Experimental Rhinology and Immunology of the Nose (SERIN 2017) Düsseldorf, Germany. 30 March–01 April 2017 Published: 23 August 2017 © 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. Oral Abstract Session 1: Rhinitis O01 Long‐term follow‐up of local allergic rhinitis patients Ibon Eguiluz Gracia, Carmen Rondón, Paloma Campo, Ana Prieto, Lina Mayorga, Luisa Galindo, Ana Molina, Miguel Blanca, Maria Jose Torres Hospital Regional Universitario de Malaga and IBIMA, Málaga, Spain Correspondence: Ibon Eguiluz Gracia iboneguiluz@gmail.com Clinical and Translational Allergy 2017, 7(Suppl 3):O0


Introduction:
Recent studies suggest that a specific respiratory microbiome is associated with an increased risk of viral infection and that in turn these infections influence the composition of the resident microbiota. A commonly found virus that causes infection in preschool children is Rhinovirus (RV), wheeze is an important symptom. Children with a symptomatic RV infection have an increased risk to develop persistent wheeze and asthma.
If children with a RV infection have a specific respiratory microbiome this may hold implications for early diagnosis of asthma and novel therapeutic approaches aimed at restoring microbial dysbiosis. Methods: We aimed to study this by comparing the nasal microbiome of children with physician confirmed wheeze, during acute symptoms and after recovery, to symptomatic controls with non-wheezing respiratory illness and asymptomatic healthy controls. Specificity of our outcomes for RV was determined by matched analysis in RV negative (RV-) children. As part of the EUROPA-study children were visited within 8 h of exhibiting respiratory symptoms and again upon recovery for assessment of symptoms and nasal swab collection. Swabs were tested by qPCR for 14 respiratory viruses. Bacterial microbiota analysis was done by IS-pro, a 16S-23S PCR-based bacterial profiling technique. Relative abundance and Shannon diversity index were compared between groups. Results: 160 pre-school children were included in the study. RV induced wheeze had the highest normalized total bacterial abundance (mean ± SEM; 0.95 ± 0.08) followed by symptomatic controls (0.77 ± 0.08) and asymptomatic controls (0.57 ± 0.09, p = 0.01). This increase was related to an upregulation of bacterial species belonging to the phyla of Firmicutes and Bacteroidetes of which the latter persisted after recovery from infection. None of the control subjects carried Bacteroidetes. Microbial diversity was significantly (p = 0.04) decreased in RV+ wheezing children (Median [IQR] 1.60 [0.97]) compared to RV− wheezing children (2.00 [0.75]). Conclusions: We established an increased microbial abundance and decreased microbial diversity in children with Rhinovirus induced wheeze who are at an increased risk to develop asthma. For a subset of these children this was primarily attributed to shifts in various species belonging to the phylum of Bacteroidetes. Our findings hold potential implications for the early diagnosis of asthma and novel therapeutic approaches aimed at restoring microbial dysbiosis. Keywords: Asthma, Respiratory Microbiome, Rhinovirus, Wheeze, IS-Pro Introduction: Eosinophilic mucin is a clinical relevant nasal secretion in patients with chronic rhinosinusitis (CRS) warning the surgeon of increased risks of recurrence of both nasal polyps and symptoms. While the pathophysiology leading to this thick, glue-like mucus is poorly understood, physicians are increasingly confronted to employ topical and or systemic strategies to avoid postoperative deterioration. Aim of this study was to determine a typical proteomics signature of eosinophilic mucin to improve our understanding of the underlying pathophysiology. Methods: Nasal secretions were collected from 10 healthy volunteers and compared to intraoperatively collected specimens of eosinophilic mucin. Pooled samples were analyzed using 2D-Gel PAGE, MALDI-MS and IPA analysis. Gender contributions was balanced and topical medication as well as allergy as confounding factors excluded. Results: 593 proteins were identified, of which 373 proteins were present in both pooled samples (mucin only 76, healthy lavage only 144 proteins). 303 out of 393 proteins were regulated in their expression. If a cut-off value of 1.5 was determined, 43 were induced and 40 were reprimed in eosinophilic mucin. Conclusions: Based on our results, specific proteomic changes could be observed in eosinophilic mucin enabling further insight into the specific pathophysiology leading to deterioration in this subgroup of CRS patients.

Introduction:
In nasal secretions (NS) IgE is present at very low concentrations and difficult to detect with standard methods. To evaluate the performance of a customized microarray chip (Thermo Fisher Scientific, TFS, Uppsala, Sweden) derived from the new Immu-noCAP ISAC 103 ™ for simultaneous detection of IgE to 13 components of Dermatophagoides pteronyssinus (D.pt.) and 2 of Dermatophagoides farinae (D.fa.) in serum and NS. Methods: NS were collected with both Filter Disks (FD) and Sinus Packs (SP) in 30 adult patients with allergic rhinitis to HDM and 29 adult non-allergic controls. NS samples were diluted 1:20 for FD and 1:10 for SP. Specific IgE to 13 D.pt. and 2 D.fa. components (Der p 1, rDer p 2, Der p 4, rDer p 5, rDer p 7, rDer p 10, rDer p 11, rDer p 14, rDer p 15, rDer p 18, rDer p 21, rDer p 23, clone 16; Der f 1 and Der f 2) were measured with the microarray both in serum (cut-off ≥ 0.10 ISAC standardized units [ISU]) and in NS. In NS the cut-off for IgE positivity needs to be determined. Hence, several analytical cut-off levels were tested (final cut-off reported below). Sensitivity and specificity of IgE tests in NS were calculated using the results of IgE tests in serum as a reference. Results: With the microarray, the best balance of sensitivity and specificity was achieved with a cut-off of ≥0.03 ISU in the diluted NS. With this cut-off, nasal IgE to major allergen molecules (nDer p 1, nDer f 1, rDer p 2, rDer f 2, rDer p 23) identified the mite-allergic patients with 90% sensitivity and 100% specificity. Among the mite allergic subjects, prevalences of IgE positivity to 13 of the 15 examined components were similar in serum and NS: 83-97% for Der p 2 and Der f 2; 70-87% for Der p 23; 20-33% for Der p 5 and Der p 7; and 0-23% for all others. IgE to Der p 1 and Der f 1 were detected more frequent in serum (80-83%) than in NS (37-60%). Among the non-allergic controls, no IgE to the components was detected in sera and NS, with the exception of IgE to Der p 5 that was detected (= 0.03 ISU) in 4 controls but only in their SP samples. Conclusions: An ISAC-derived microarray technique detected nasal IgE to major allergen molecules D.pt. and D.fa (nDer p 1, nDer f 1, rDer p 2, rDer f 2, rDer p 23) identifying the mite-allergic patients with 90% sensitivity and 100% specificity when referred to serum. These promising findings need confirmation in larger studies (Table 1). Clin Transl Allergy 2017, 7(Suppl 3):28 Introduction: There is a lack of published studies about the association between rhinomanometry and spirometry results. Some studies have shown a moderate correlation between spirometry parameters and other nasal objective measures such as Peak Nasal Inspiratory Flow (PNIF). We aimed to study the correlation between rhinomanometry and spirometry parameters. Methods: We included all adults (age ≥ 18 years) who performed rhinomanometry and spirometry consecutively on the same day, at CUF Porto -Instituto and Hospital from November 2010 to July 2016. When more than one rhinomanometry was performed, only the first one was included in the analysis. We included gender, age, height, rhinomanometry parameters (inspiratory and expiratory total nasal airflow, inspiratory (RAARi) and expiratory (RAARe) mean airflow resistance at a sample pressure of 150 Pa and side ratio) and spirometric parameters (forced vital capacity (FVC), forced expiratory volume in 1 s (FEV 1 ), mid-flow rate/forced expiratory flow at 25-75% of FVC (FEF25-75) and FEV 1 /FVC). Pearson's correlation was used to evaluate unadjusted correlations and partial correlations were used to adjust parameters for age, gender and height. Results: A total of 971 adults were included, 623 (64%) females, with a mean (sd) height of 166.0 (9.0) cm and age of 38.3 (14.1) years Clin Transl Allergy 2017, 7(Suppl 3):28 (min-max: 18-80). Correlations between spirometry and rhinomanometry variables are presented in Table 1. The correlations between FEV 1 /FVC and either inspiratory total nasal airflow or mean RAARi, were the only ones with statistical significance (r = −0.083, p = 0.016 and r = 0.011, p = 0.039, respectively). After adjusting for age, gender and height, no statistically significant associations were found between the parameters. Introduction: Infrared thermography is a diagnostic tool in more medical disciplines. It detects temperature changes in the infrared spectrum range and was originally designed to test the heating qualities of buildings. Potential benefits in allergology and ENT are not widely evaluated. We asked if this method can give us an objective information about a patient during nasal provocation test.
Methods: As it is difficult to detect always the same point in the face during repeating measurements, a bounding box to detect the face of a patient was designed. Forehead, nasal, cheek and chin facial areas were defined. Results: We present one example of our software in the evaluation of facial infrared thermography. This method detects temperature changes in different areas of the face. The nasal area was the most sensitive one in the detection of temperature change during nasal provocation test followed by the cheek area. Conclusions: Facial infrared thermography might be-besides other important parametres like nasal symptome score and active anterior rhinomanometry-an objective, non-invasive and feasible tool in the assessment of allergic response during nasal provocation test. Introduction: Condyloma acuminatum is a venereal diasese transmitted by the human papillomavirus (HPV). Generally, it is a benign entity but carcinomatous change has been reported in anogenital area. The malignant transformation is associated with the immunocompromised status,especially HIV. Condyloma acuminatum is uncommonly identified in the nasal cavity, the malignant transformation is extremely rare. Methods: A mid-aged patient with diabetes mellitus and psoriasis had suffered from progressive right nasal obstruction with epistaxis for six months. Physical examination revealed a cauliflower-like lesion over the right nasal vestibule, expanding to the septum. Due to this patient had history of penile condyloma acuminatum, biopsy was done and sent hc2 high-risk-HPV DNA testing. The results was compatible with condyloma acuminatum and negative for high-risk-HPV infection. Results: Eradication surgery was performed. Necrotic tissue with pus content was buried in the lesion. This mass was completely excised eventually. The formal histopathology reported condyloma acuminatum with focal invasive squamous cell carcinoma. We suggested this patient receiving adjuvant radiotherapy or re-operation because there was no planed safe margin in advance, and the examinations for HIV and syphilis were also advised. But the patient refused and lost of follow-up thereafter. Conclusions: Human papillomavirus types 16 and 18 are found in up to 90% of patients with cervical carcinoma, however HPV type 6 and HPV type 11 are the main factor in developing giant condyloma acuminatum, which is reported 56% incidence of malignant transformation. Abscesses and fistulas are more common in lesions as described in our case. Immunosuppression, coexisting HIV infection, and unhygienic conditions play a role in malignant transformation, therefore AIDS togethers with other venereal diseases should be examined. Electrocautery or laser total excision could be applied for treatment. Non-resectable lesions, radiotherapy could be applied either alone or along with chemotherapy. In conclusion, condyloma acuminatum with invasive squamous cell carcinoma is rarely found in the nasal cavity, more cases should be obtained for more comprehensive understanding. Introduction: One of the most common problems presenting among physicians are rhinitis symptoms. Challenges in the diagnosis results from the factor that differential diagnosis is extensive and sometimes symptoms of allergic, nonallergic, and mixed rhinitis are often indistinguishable. Searching in the literature there is a clear consensus on the classification but is difficult to found one that include and ordered all the different etiologies. It is also difficult to found an specific diagnosis algorithm for these diseases. Methods: After checking over and try to reunite different consensus made in allergy an ENT for classification and diagnosis of rhinitis we propose Tables 1, 2 with both of these sections. Results

CHRONIC RHINOSINUSITIS
Conclusions: When approaching rhinitis, diagnostic challenge is to determine the etiology, specifically whether it is allergic, nonallergic, or perhaps an overlap of both conditions. With these proposal tables it could be more easy to differentiate and lead to a correct diagnosis. Introduction: Most allergic rhinitis (AR) patients attending clinic have moderate/severe persistent disease. Meda Pharma's AzeFlu (MP-AzeFlu) comprises intranasal azelastine hydrochloride, fluticasone propionate and a novel formulation, in a single device. Its reallife effectiveness has been established in AR during 14 days, but its effectiveness in those with persistent AR (PER) over the longer term is unknown. We assessed the effectiveness of MP-AzeFlu in routine clinical practice in PER patients, using the ARIA-endorsed language of AR control (i.e. visual analogue scale (VAS)). A VAS score cut-off of 50 mm is recommended to assess control and guide treatment decisions. Methods: 428 patients (≥12 yrs old) with moderate/severe PER were recruited into 3, prospective, non-interventional studies in Austria (n = 214), Ireland (n = 53) and Sweden (n = 161). MP-AzeFlu was prescribed according to label. Patients assessed symptom severity using a VAS from 0 mm (not at all bothersome) to 100 mm (very bothersome) in the AM prior to MP-AzeFlu use, on Days 0, 1, 3, 7, 14, 21, 28, 35 and 42. An endoscopy was performed in the Irish study on Days 0 and 28. Symptoms of 'oedema' , 'discharge' and 'redness' were scored on a 3-point scale for both nostrils (max score = 12). Results: Patients treated with MP-AzeFlu (1 spray/nostril bd; daily doses: AZE = 548 μg;FP = 200 μg) had a VAS score reduction from 59.0 mm (SD 25.0) at Day 0 to 28.8 mm (SD 23.2) at Day 28 (p < 0.0001) and 23.2 mm (SD 21.5) on Day 42 (p < 0.0001), a reduction of 35.8 mm. This reduction was rapid, with statistical significance vs baseline noted from Day 1 (p < 0.0001), and was consistent irrespective of phenotype, patient age, and disease severity. On average MP-AzeFlu-treated patients achieved the 50 mm VAS score cut-off before Day 3. Total endoscopy score reduced from 7.5 (SD 3.1) at baseline to 3.5 (SD 2.5) at Day 28. The % of patients with severe oedema reduced from 53.1% at baseline to 3.8% at Day 28. A similar reduction in the incidence of thick/mucusy discharge (28.3-4.8%) and severe redness (34.9-0%) was also observed. Conclusions: MP-AzeFlu provides effective and rapid PER control in a real-world pan-European setting assessed by VAS. Symptom improvement was noted from Day 1, sustained for 42 days and was associated with improvement in mucosal appearance after 28 days. Keywords: MP-AzeFlu, Dymista, Azelastine, Fluticasone *MP-AzeFlu, a registered trademark of Meda AB, is marketed in the U.S. as Dymista ® , a registered trademark of Meda Pharma Inc., both Mylan Companies. monotherapy in patients with allergic rhinitis and chronic rhinitis. MP-AzeFlu has previously shown greater antiinflammatory potency than FP and AZE in an in vitro model of eosinophilic inflammation. Our aim was to compare the effect of MP-AzeFlu, FP, and AZE on antiinflammatory gene transactivation and proinflammatory mediator inhibition in sinonasal cultured fibroblasts from nasal mucosa (NM) and chronic rhinosinusitis with nasal polyps (CRSwNP). Methods: NM and CRSwNP fibroblast cultures (n = 6) were incubated with serial dilutions of MP-AzeFlu (1:10 2 -1:10 4 ) or FP or AZE (equivalent dilutions) for 2 to 24 h with/without 1 ng/ml IL-1b. Glucocorticoid-induced leucine zipper (GILZ), mitogen-activated protein kinase phosphatase-1 (MKP-1), and COX-2 gene (RT-PCR) and IL-8 and GM-CSF protein (ELISA) expression was analysed. Results are expressed as fold-increase or % inhibition over control (mean ± SEM). Results: MP-AzeFlu and FP (all dilutions, P < 0.05) and AZE (1:10 2 , P < 0.05) increased GILZ and MKP-1 gene expression in NM and CRSwNP fibroblasts. MP-AzeFlu (1:10 2 ) had an overadditive effect on GILZ expression vs FP or AZE and on MKP-1 expression vs AZE (P < 0.05) ( Table 1). MP-AzeFlu and FP provoked a similar inhibition of IL-1b-induced COX-2, IL-8, and GM-CSF in NM and CRSwNP fibroblasts (all dilutions, P < 0.05) but greater than AZE, which had no inhibitory effect. We have previously shown that VEGF is overexpressed in the epithelium of hyperplastic nasal polyps in patients with chronic rhinosinusitis with nasal polyposis (CRSwNP). We hypothesized that the HIF family of transcription factors function to regulate this process. Methods: We used human primary nasal epithelial cells grown in submerged culture (PNEC) and whole human polyp tissue in air liquid interface culture to examine to examine HIF and VEGF expression by immunohistochemistry, flow cytometry and realtime PCR analysis. Function was assessed by exposure to known pharmacologic inhibitor digoxin and specific siRNA knockdown of HIF on cell growth and apoptosis by DNA binding assay of PNEC and TUNEL assay of polyp tissue, respectively. Results: We now report that the HIF family of transcription factors, both HIF1-alpha (HIF1α) and HIF2-alpha (HIF2α), which known regulators of expression of VEGF, are highly expressed in PNEC from CRSwNP subjects, as compared to normal control subjects when analyzed by flow cytometry of PNEC and immunohistochemistry of sinonasal surgical tissue. PNEC from CRSwNP patients express high constitutive levels of mRNA of HIF1α and HIF2α. Inhibition of HIF1α by cardiac glycoside digoxin results in 65% inhibition of PNEC growth rates analyzed by DNA binding assay (p < 0.001, n = 5). Further studies show that siRNA knockdown of HIF1α results in inhibition of VEGF mRNA expression by 75% (p < 0.01, n = 3) and subsequent 55% inhibition of PNEC growth rate as measured by DNA binding assay (p < 0.01, n = 3). Exposure of whole human nasal polyps in ex vivo culture to HIF1α inhibitor digoxin or VEGF co-receptor inhibitor anti-neuropi-lin1 antibody results in 90% (p < 0.01, n = 5) and twofold (p < 0.005, n = 5) increase in apoptosis respectively of PNEC as assessed by TUNEL assay. Introduction: Two major clinical phenotypes of chronic rhinosinusitis (CRS), with (CRSwNP) and without nasal polyps (CRSsNP) may show different regulation of inflammation, interplay between T cell subsets, remodelling, response to microbiome and association with predisposing comorbidities, like allergy, asthma and aspirin intolerance. IL-7 and IL-15 belong to common gamma chain cytokines, and are essentially involved in T cell homeostasis and proliferation. Several papers indicated that they might be involved in the immune response in asthma, however, their role in CRS was not evaluated. Our aim was to analyze interaction between IL-7 and IL-15 related to phenotypes and comorbidities.

Methods:
The study included 50 patients(both gender, age 18-60 years, 32 with CRSsNP, and 18 with CRSwNP), with CRS according to the EPOS criteria, who were submitted to endoscopic sinus surgery. Ethmoid sinus mucosa or nasal polyps were collected at surgery and immediately tissue homogenates were prepared and incubated with protease inhibitor. Assays (R&D System, Quantikine ELISA, UK) for interleukins IL-4, IL-5, IL-7, IL-15 and for interferon gamma (IFN-γ) employs the quantitative sandwich enzyme immunoassay technique. Tissue homogenates (n = 70) were analysed according to the manufacurer's instructions (29 from non-allergic CRSsNP, 17 from allergic asthma patients, 11 from non-allergic asthma, 9 from allergic non-asthmatic CRSsNP and 4 from aspirin intolerant CRSwNP). Polyp and ethmoid sinus mucosa samples were taken in CRSwNP, and only ethmoid sinus mucosa from patients with CRSsNP, respectively, from the worse side according to imaging. Results: Results. IL-5 was significantly higher (mean 38.69 vs. 9.47 pg/ mL respectively), but IL-7 (0.1397 vs. 0.2638 pg/mL, respectively), and .71 pg/mL, respectively) significantly lower in samples from CRSwNP, and these differences were also significant when asthmatic were compared to non-asthmatic, irrespective of phenotype and sensitization. Interestingly, IFN-γ was significantly higher in patients with allergic sensitization, while IL-4 was significantly higher in patients with ASA intolerance. IL-15 significantly correlated with IL-7 (rho 0.44), IL-4 (rho 0.31) and IFN-γ significantly with IL-4 (rho 0.51). Conclusions: IL-7 and IL-15 are significantly upregulated in CRSsNP compared to CRSwNP and in non-asthmatics compared to asthmatics. Keywords: IL-7, IL-15, Chronic Rhinosinusitis, Nasal Polyps, T Cell Subset Introduction: Acute and chronic rhinosinusitis affect everyone several times per year and 11% of the EU population, respectively. Methods: Double-blind randomized placebo-controlled trials. Results: Recently, the herbal drug BNO 1016 showed evidence for a significant and clinically relevant reduction of symptoms and increase of quality of life. In an analysis of pooled data of two similar randomized placebo-controlled clinical trials including 589 patients, the Major Symptom Score (MSS) improved from 10.02 ± 1.61 to 2.47 ± 2.55 for BNO 1016 and from 9.87 ± 1.52 to 3.63 ± 3.63 for placebo; the difference between treatment groups at end of therapy (1.16 score point ± [SD]) was statistically significant in favor of BNO 1016 (p < 0.0001); patient-assessed quality of life also improved significantly (p = 0.0015) versus placebo. The results were confirmed by ultrasonography. In summary, the herbal dry extract BNO 1016 is efficacious and well tolerated in patients with acute viral rhinosinusitis. The objective of a recent clinical trial therefore was to assess the efficacy, safety and tolerability of dry extract BNO 1016 in patients with chronic rhinosinusitis. 927 patients suffering from chronic rhinosinusitis (>12 weeks and 6 ≤ MSS ≤ 12) were enrolled in a randomised placebocontrolled trial with a treatment period of 12 weeks. The primary endpoint was the mean Major Symptom Score (MSS) over week 8 and week 12 compared to placebo. Secondary endpoint included further MSS related calculations and responder rates over time. Finally, safety and tolerability were evaluated. Although the results of the secondary endpoints showed a clear trend towards superior efficacy, BNO 1016 was not superior over placebo regarding the primary endpoint. Additional post hoc sensitivity analyses in patients with a baseline MSS ≥ 10, duration of disease > 1 year and diagnosed by specialists in otorhinolaryngology indicated that those patients significantly benefited from BNO Clin Transl Allergy 2017, 7(Suppl 3):28 1016. Therapy was superior for the primary endpoint analysis. Patients were less impaired with respect to work and daily activities. A good safety and tolerability of BNO 1016 was assured in all patients. Conclusions: Rhinosinusitis, a frequent cause of disease and associated sequalae, still remains a challenge for treatment. BNO 1016 can safely be administered in patients with acute and chronic rhinosinusitis and offers a suitable treatment option. Introduction: Aspirin exacerbated respiratory disease (AERD) is characterized by acute aspirin-triggered respiratory distress, chronic asthma and nasal polyps that are often refractory to steroid treatment. AERD patients show characteristic abnormalities in lipid mediator (eicosanoid) metabolism and signaling, but the mechanisms that discriminate nasal polyp development in AERD and aspirin tolerant chronic rhinosinusitis with nasal polyps (AT CRSwNP) remain obscure. We thus compared eicosanoid profiles in nasal polyps from AERD and AT CRSwNP patients and analyzed the expression of factors involved in airway remodeling. Methods: Nasal polyp tissues (n = 10 AT CRSwNP, n = 4 AERD) were obtained from patients undergoing functional sinus surgery. All patients were treated with systemic steroids preoperatively. Expression of eicosanoid pathway proteins and regulatory factors in nasal polyp tissues or healthy turbinates (n = 3) was studied by immunofluorescence and immunohistochemistry followed by automated image analysis. Supernatants from overnight cultures of nasal polyps were processed and subjected to lipid mediator analysis by LC-MS/MS or immunoassay. Results: Nasal polyp tissues from steroid treated AT CRSwNP and AERD patients expressed high levels of leukotriene (LT) biosynthetic enzymes (5-lipoxygenase, Leukotriene C4 synthase) despite low levels of infiltrating granulocytes. LT enzymes were abundant in the nasal polyp epithelium and in macrophages and all nasal polyp tissues released significant amounts of LTs. The overall eicosanoid profile was similar in nasal polyps from AT CRSwNP and AERD patients with a tendency for increased thromboxane production by AERD tissues. In contrast AERD tissues expressed significantly lower levels of the enzymes transglutaminase 2 (TG2) and microsomal prostaglandin E2 synthase (mPGES-1). TG2 was particularly abundant in tissues from house dust mite allergic patients. Conclusions: Our findings identify TG2 as a potential mechanism that discriminates nasal polyp endotypes. We further show that TG2 expression and LT production in nasal polyp epithelium are resistant to steroid treatment. Thus, targeting TG2 might be promising in allergic CRSwNP patients and novel strategies targeting steroid resistant lipid mediator abnormalities in nasal polyps are urgently needed. Keywords: AERD, Allergy, Eicosanoids, Nasal Polyps, Transglutaminase 2

P12 Effects of Cytokine secretion from nasal polyps on peripheral lymphocytes in a co-culture system
Introduction: T cell subpopulations in nasal polyps differ from peripheral lymphocytes in patients with CRSwNP. The direct influence of the nasal polypoid tissue by secretion of cytokines on the differentiation or activation of T cells still remains unclear. To study the effects of polypoid tissue on peripheral lymphocytes in vitro, we developed a co-culture system with nasal polyp tissue and peripheral lymphocytes to directly measure these interactions. Additionally, we assessed the cytokine secretion by polyp tissue, which may induce T cell responses in this system. Methods: Tissue and blood samples were collected from 10 patients undergoing nasal sinus surgery. Polypoid tissue was cultured under air-liquid interface conditions. Afterwards, CD3/CD28 activated PBMC of the same patients were added using cytokine free medium. After 3 days, lymphocytes were separated and analyzed by multicolor flow cytometry. Monoculture PBMC served as control group. Additionally, cytokine secretion of the polyp tissue was measured using a human Th1/Th2/Th17 antibody array. Results: There was a significantly higher amount of CD4 + and CD8 + T cells in the co-cultured system than in PBMC alone. Terminal differentiated CD8 + T cells were significantly increased, while central memory CD8 + T cells significantly decreased in the co-culture. HLA-DR was downregulated in co-cultured CD3 + lymphocytes. Conventional memory CD4 + T cells significantly increased and resting regulatory T cells significantly decreased in the co-cultured system. Cytokine analysis showed a secretion of IL-6, GM-CSF and MIP-3.

Conclusions:
It could be demonstrated that nasal polypoid tissue has an effect on the PBMC phenotype. The secretion of pro-inflammatory cytokines may play a regulatory role in this context. Interestingly, the clinically known downregulation of HLA-DR in CD3 + lymphocytes was significantly reproducible in vitro. The modulating effect of the polypoid tissue on the activation of lymphocytes must be further investigated in order to determinate its impact on this disease. Keywords: Chronic Rhinosinusitis With Nasal Polyps, T Cell Subsets, Co-Culture System Clin Transl Allergy 2017, 7(Suppl 3):28 for the final diagnosis of EGPA. The aim of the study was to develop a new diagnostic tool to support the diagnostic and prognostic work-up of EGPA patients with the possibility to evaluate extracellular mediators in nasal secretion. Methods: Nasal secretions were gained from 40 patients of which 20 EGPA, 10 suspected EGPA and 10 controls after positioning the cotton pieces in the nasal middle meatus where they were left for 10 min. Subsequently the cotton pieces were put in 5 ml of saline solution and left in ice until processing. The liquid obtained from the squeezing of the cottons was centrifuged two times at 1600 RPM for 10 min. Supernatant was frozen after first centrifugation. Trypan Blue was added to 10 μl of sample to calculate the number of cells and their vitality. Finally, 100 μl of sample with containing 1.5-2.00 × 10 5 cells were centrifuged with Cytospin at 300 RPM for 15 min. The slides were stained with May Grunwald/Giemsa and cell population analysed at microscope with objective oil immersion 40× of magnification. The remaining cells were frozen or cultured. Results: The mean number of cells obtained after centrifugation was 2.00-4.08 × 10 6 cells/ml for EGPA patients and 6.42-8.62 × 10 5 cells/ ml for control group and suspected EGPA. Cytologic analysis allowed us to count an increase in the percentage of eosinophils in EGPA patients with active ENT disease and in some of the patients with a suspect of EGPA, where this percentage correlated with histologic evidence of disease. Conclusions: This method seems preliminarily a reliable cytologic examination allowing fresh cells isolation and analysis of extracellular mediators. Compared to the direct slither of the mucus on the surface of the glass, cytospin significantly reduced any dye accumulation. Further studies are ongoing to support the described method, in order to confirm the cytologic diagnostic reliability to identify prognostic biomarkers of EGPA. Introduction: Type I interferons (IFN-I) response has been implicated in the eosinophilic inflammation besides its antiviral function. This study aimed to investigate the role of IFN-I response in the pathogenesis of eosinophilic chronic rhinosinusitis (ECRS). Methods: The expressions of IFN-I and IFN-I receptor (IFNAR1) in sinonasal tissue from controls and patients with CRS with nasal polyp (NP) were measured using real time-PCR, ELISA, and/or immunohistochemistry. The levels of CCL11/eotaxin-1, IL-5, and IL-13 in sinonasal tissue of human subjects were determined by ELISA. ECRS in the wild type (WT) and IFNAR1 knockout (Ifnar1 −/− ) mice was induced by intranasal challenge of aspergillus protease plus ovalbumin (OVA). Stromal cells cultured from NP tissue were stimulated by exogenous IFN-β and their CCL11/eotaxin-1 production was measured by ELISA. Results: IFN-β, IFNAR1, IL-5, IL-13, and CCL11 expressions were increased in NP tissues from ECRS compared with uncinated process mucosa from controls. IFN-β levels positively correlated with IL-13 and CCL11 levels. The histological severity of Aspergillus protease plus OVA induced-ECRS was less in Ifnar1 −/− than in WT mice. The levels of IL-4, IL-5, and CCL11 in the nasal lavage fluid of Ifnar1 −/− mice were significantly lower than those in WT mice. The serum total IgE levels in Ifnar1 −/− mice were also lower than those in WT mice. NP stromal cells produced significantly greater amount of CCL11/eotaxin-1 by concentration-dependent stimulation of IFN-β. Conclusions: Our results showed that IFN-I response was up-regulated in the NP tissues from ECRS, IFN-β increased CCL11/eotaxin production in the NP stromal cell culture model, and development of ECRS was inhibited by deficiency in IFN-I signaling. Therefore, increased IFN-I response in the sinonasal mucosa may underlie in the pathogenesis of ECRS. Keywords: Chronic Rhinosinusitis, Type I Interferon, Eosinophil Introduction: Chronic rhinosinusitis with nasal polyps (CRSwNP) affects 4% of the total population (C.A. Akdis, 2013). The prevalence of CRSwNP in Russian Federation (RF) is 1.5% or 4.9 per 10,000 population (Lopatin A, 2014). The ratio and number of neutrophils and eosinophils in the vast majority of nasal polyps (NP) are diverse. According to the data from research neutrophilic types of adult bilateral polyps do occur, predominantly in Asian subjects and some populations in North America (Zhang N, 2006). Russia is a multiethnic country that separates Europe and Asia. It is of interest as a target for epidemiological studies. The aim was to explore the histology of NP biopsies obtained from operated patients with CRSwNP living in different regions of RF. Methods: NP samples obtained from 152 patients during polypectomy were assaying histologically to identify neutrophils and eosinophils count and ratio of eosinophils to neutrophils (ENR) in biopsies. There were 10-11 samples of polyp's tissue from 15 ENT hospitals from different regions of the country. There was equal quantity of participants of 12 nationalites belonging to Caucasian or Mongoloid race. All patients completed a questionnaire including data about symptoms, nationality, comorbidities, atopic reactions and drug hypersensitivity. Results: There was predominance of eosinophil's infiltration (>90% of all cells) in all NP samples. There were no significance between clinical symptoms and NER. The NP with predominant eosinophil's count (ENR median total = 6.8) were isolated from Siberian and Volzhsky region habitants (Tomsk ENR = 23.3, Irkutsk = 9.14, Nizhniy Novgorod = 11.3). There were no significant differences in ENR in patients of different nationalities. Moreover the predominance of eosinophilic infiltration in Russian (Caucasoid) were revealed in 69.5% of cases versus patients with other nationalities patients (83%). In most biopses from Russian patients ENR = 6.0 in comparison to others ENR = 6.9. Conclusions: There're some hypotheses about triggering factors of CRSwNP. The most popular one is the influence of microbial agents (C. Bachert). Probably the features of infectious and parasitic diseases (zoonoses, insect bites, ticks, infected fish, etc.) and climate factors are cause of debut of formation of polyps. Russia is excellent model for epidemiological studies that could reveal these factors. Keywords: Nasal Polyp, CRSwNP, Eosinophils, Endotypes Introduction: Is concurrent eosinophilic oesophagitis and sinonasal polyposis a pure coincidence? Or, do they share common pathogenesis? We report a case series of four subjects with both conditions managed at a local allergy clinic in Adelaide, South Australia, Australia, highlighting certain features common to both. Methods: N/A. Results: All four subjects ranged from age 50 to 61 at the time of presentation, seeking treatment for chronic nasal symptoms, which were managed predominantly with intranasal corticosteroids. All four were asthmatic and have had intranasal surgery in the past, with two of them having had four surgeries to date. Results of polyp biopsy were not available. Peripheral eosinophilia was characteristic of all four, ranging from 0.6 to 1.56 × 10 9 . Background of atopy was Clin Transl Allergy 2017, 7(Suppl 3):28 further evaluated with skin prick testing with aeroallergens and food allergens with no concordant findings noted across all four subjects. Nasal symptoms preceded oesophageal symptoms by 2 to 3 years. All subjects had oesophageal biopsy confirming eosinophilic oesophagitis. Three subjects were aspirin sensitive. Aspirin or sodium salicylate desensitisation was tried without success. Two subjects responded to low salicylate diet with the remaining two not yet trialled on such. Three subjects responded symptomatically to montelukast. Conclusions: Is there a single factor driving the pathomechanistic pathway e.g. IL-5? Nasal polyposis occurs frequently in patients with intolerance to salicylate acid and those with allergic fungal sinusitis. It has been demonstrated that IL-5 has a key role in the pathophysiology of eosinophil dominated polyps. Additionally, adhesion molecules VCAM-1 is recognised to play a role in the extravasation of eosinophils into nasal polyps and cytokines such as IL-3, IL-4 and TNF-a can induce VCAM-1 expression in microvascular endothelium from the polyps. Future planned testing includes eosinophil activation markers on flow cytometry, along with serum levels of cytokines mentioned. Further questions then arise for the management of future patients, especially in regards to the eosinophilic oesophagitis component. Introduction: Traditionally, Draf III procedure is considered for accessing the refractory pathogens inside the frontal sinus. In case of confined, predominantly unilateral lesions, Draf IIb procedure also provides wide access to the frontal sinus. This approach can be extended without destruction of the contralateral frontal sinus drainage pathway. Methods: We report on a patient suffering from left frontal pain and nasal obstruction for 6 months. He received examination in our department, and fiberscopy revealed polyposis. Computed tomography displayed the over-pneumatized right frontal sinus pushing intersinus septum toward to left side and causing narrowed left frontal space. We decided to performe Draf IIb.2 procedure. Results: Once the frontal recess is identified, we started using the irrigated RAD curved burr (3.6 mm 55°) to remove partial anterior-superior portion of middle turbinate and frontal beak. After identifying intersinus septum, we removed its lower part and formed a drainage pathway through rightl frontal sinus. There were no sequelas and patient felt well during follow-up. Conclusions: In case of confined, unilateral frontal lesions, less destructive, limited approaches, defined as extended Draf IIb, can be applied without disturbing the contralateral frontal sinus drainage pathway according to the literatures. Gotlib et al. have classified extended Draf IIb procedure into 3 subtypes; Extended Draf IIb.2 (or mini-Lothrop) refers to Draf IIb procedure with removal of the lower intersinus septum. This technique is applicable for bilateral frontal lesions with one side limiting in frontal sinus or unilateral frontal sinus disease with intersinus septum deviation towards the lesion. It can minimiz destruction of the contralateral frontal sinus drainage pathway when comparing to Draf III. The nasal septum can be kept intact as compared to Draf IIb.1 and Draf IIb.3 procedures. Keywords: Draf IIb Procedure Introduction: Birch pollen allergic rhinitis (AR) is one of the common allergic diseases in Europe. Little knowledge exists on changes in nasal epithelial transcriptome during allergen exposure or allergen specific immunotherapy. The aim of this study was to evaluate the seasonal and immunotherapy-related alterations in nasal epithelial transcriptome in subjects with or without birch pollen allergic rhinitis (AR). Methods: The study subjects were healthy nonsmoking adults with or without birch pollen AR (N = 11). We used the RNA sequencing to demonstrate the alterations in transcriptome of study subjects. The whole study spanned on two years i.e. 4 seasons, where half of the AR patients started the immunotherapy during the second year. We used edgeR for differential expression analysis and R environment for further Bioinformatics. Gene Ontology (GO) and KEGG pathway enrichment was done using DAVID (Database for Annotation, Visualization, and Integrated Discovery). Results: We found a significant change in gene expression of the nasal epithelial cells in AR patients not having the immunotherapy compared to healthy controls. We found immunity related GO terms up or down -regulated in these patients. We also observed variations in biological processes of IL-2 production, cytokines production, regulation of lymphocyte and leukocyte differentiation. Immunotherapy induced a down regulation of GO terms related to immune response in AR patients. Introduction: The effect of allergen-specific immunotherapy (IT) on basophils remains incompletely understood. Several studies have shown an early suppression of basophil activation, and a few studies have reported long-term suppression in response to IT. These studies involved venom, grass, birch or cat IT. To our knowledge, no previous studies investigated the effect on basophil activation in the context of house dust mite (HDM) IT. Objectives: To study the effect of HDM IT on allergen-stimulated basophil activation and to evaluate its relation with clinical response to treatment. Methods: Patients with HDM allergic rhinitis undergoing conventional HDM subcutaneous IT (HDM SCIT, n = 23) and patients not