SUNDAY , SEPTEMBER 8 TH 2013 65 . Biomarkers in asthma and COPD

printing supported by . Visit Pfizer Vaccines at Stand Q.01 52s SUNDAY, SEPTEMBER 8TH 2013 395 Altered infl ammasome activity in bacterial acute exacerbations of COPD (AECOPD) Efrossini Dima1,2, Matina Kardara2, Harilenna Giannakopoulou1, Charis Roussos2, Nikolaos Koulouris1, Nikoletta Rovina1,2. 11st Department of Pulmonary Medicine, “Sotiria” Hospital, Athens Medical School, Athens, Greece; 2”M. Simos” Laboratories, Pulmonary and Critical Care Department, Evangelismos Hospital, University of Athens, Athens, Greece Infl ammasome and its products, as part of the innate immune system, can be triggered to assist in defence against invading pathogens. We have previously shown increased levels of IL-18 in induced sputum of stable COPD patients, which were decreased in acute exacerbations of COPD (AECOPD), implying a possible dysregulation of infl ammasome in AECOPD. Aim of this study was to assess the infl ammasome activity in AECOPD in the case of proven bacterial infections versus AECOPD where only bacterial colonization was proved. 30 patients hospitalized for an infectious AECOPD according to Anthonisen’s criteria were included in the study. We examined the infl ammatory properties of induced sputum and assessed bacterial infection using PCR. IL-18, caspase-1, TLR-2, and IL-1b were measured in induced sputum and serum by immunosorbent analysis (ELISA). Immunocytochemistry of IL-18 expression in sputum cells was performed using a mouse monoclonal IL-18 antibody. IL-18 levels in sputum were found signifi cantly lower in AECOPD caused by a pathogen compared to colonized AECOPD (207 pg/ml (range 47-2301) vs 420 pg/ml (58-1201), p=0.05). Similarly, although non statistically signifi cantly, decreased levels of caspase-1 and TLR2 were found in infectious versus colonized AECOPD (1.6 pg/ml (1.22-19) vs 3.6 pg/ml (1.27-117), p>0.05). IL-1b was statistically signifi cantly increased in induced sputum of infectious AECOPD (539 pg/ml (1.5-973) vs 88 (2.9-989), p<0.05). Positive staining of IL-18 was observed in macrophages in immunocytochemistry. Our data show that in bacterial AECOPD there may be a dysregulated activation of infl ammasome mediating IL-18 production. 396 Local and systemic infl ammation in chronic obstructive pulmonary disease (COPD) Jie Ji1, Ida von Schéele1, Barbro Dahlén2, Jan Bergström3, Bo Billing2, AnnSofi e Lantz2, Kejll Larsson1, Lena Palmberg1. 1Lung and Allergy Research, Institute of Environmental Medicine, Karolinska Institutet, Stockholm, Sweden; 2Lung and Allergy Research, Department of Medicine, Huddinge (MedH), Karolinska Institutet, Stockholm, Sweden; 3Department of Dental Medicine, Karolinska Institutet, Stockholm, Sweden Background The aim of the study was to explore to what extent local infl ammatory processes in the mouth (saliva, clinical assessment of periodontitis) and the respiratory tract (sputum, bronchoalveolar lavage (BAL), lung function) are associated with systemic infl ammatory responses (blood) in smokers with and without COPD. Method Healthy controls (n=23), smokers with (n=28) and without (n=29) COPD performed spirometry and dental examinations. Saliva, induced sputum, BAL fl uid and serum were collected. Infl ammatory mediators were measured using ELISA. Soluble and cell bound tumor necrosis factor receptors (TNFR) in sputum, BAL fl uid and serum were detected by fl ow cytometry. The mRNAexpression of tumor necrosis factor- (TNF-) and its receptors on BAL macrophage were analyzed by real-time PCR. Result A negative correlation between lung function and saliva IL-8/MMP-9 was found in smokers with COPD (p<0.01). There were positive correlations between these mediators (IL-8/MMP-9) in saliva and periodontitis as assessed by bleeding index in non-smokers (p<0.01). Sputum IL-6 and IL-8 were signifi cantly positively correlated with soluble TNFRs (sTNFRs) in non-smokers (p<0.01) and with sTNFR2 in smokers with COPD (p<0.01). There was a close positive correlation between soluble TNFR1 and TNFR2 receptors in sputum, BAL and serum in all groups (p<0.01). Conclusion Infl ammatory markers in saliva, which is easy to collect, seem to refl ect disease severity in COPD patients. Shedding of TNFR is similarly regulated locally and systemically, both in healthy subjects and in smokers, irrespective of airfl ow limitation. 397 Pro-surfactant protein B, a promising BAL biomarker of COPD progression in heavy smokers, is increased by budesonide/formoterol shortterm therapy Anthony Tam1, Soo Jung Um1, Harvey Coxson2, Stephen Lam1, Shu Man1, Don Sin1. 1Medicine, University of British Columbia, Vancouver, BC, Canada; 2Radiology, University of British Columbia, Vancouver, BC, Canada Rationale  Aim: Reduced levels of surfactants in lung and bronchoalveolar lavage fl uid (BAL) generally signal disease progression. The aim of this study was to determine the effects of Symbicort® therapy on the BAL levels of prosurfactant protein B (pro-SPB) and other biomarkers in heavy smokers with or without COPD. Methods: We recruited 37 heavy smokers (3 current and 34 former; 30 packyears), age 65±6 years (mean±SD), free of exacerbations for  4 weeks, with FEV1 of 73.1±18.3% predicted and FEV1/FVC ratio 66.3±9.4% (clinical trials. gov: NCT00569712). COPD was defi ned as FEV1/FVC <70%. BAL was obtained at baseline and after 4 weeks of Symbicort Turbuhaler® 400/12 mcg (budesonide/formoterol) BID therapy. Lung-predominant proteins: pro-SPB, surfactant protein D (SP-D) and Club Cell Secretory Protein (CCSP)-16 were measured in BAL supernatants. Results: Symbicort therapy signifi cantly increased pro-SPB levels in BAL (geometric mean±SD: 322±619 versus 268±394 ng/ml; p=0.0166). The pro-SPB levels (but none of the other BAL biomarkers) were signifi cantly related to lung function expressed by FEV1% of predicted (Spearman rho=0.36; p=0.026) and FEV1/FVC ratio (rho=0.51; p=0.0013), and to the levels of SP-D (rho=0.43; p=0.0073) and CCSP-16 (rho=0.54; p=0.0005), and to body mass index (rho=0.46; p=0.0043). Conclusions: In the current and former heavy smokers, pro-SPB levels in BAL were positively related to lung function and signifi cantly increased by 4 weeks therapy with Symbicort. Pro-SPB is a very promising BAL biomarker to evaluate lung function in heavy smokers and thus disease progression in COPD and other chronic airway diseases. 398 Assessment of ATP degradation in bronchoalveolar lavage fl uid Zsofi a Lazar1,2, Marisa Braun1, Anja Meyer1, Jessica Beckert1, Ildiko Horvath2, Marco Idzko1. 1Dept. Pulmonology, University Clinic Freiburg, Freiburg, Germany; 2Dept. Pulmonology, Semmelweis University, Budapest, Hungary Introduction: Extracellular adenosine triphosphate (ATP) via purinergic signalling plays a role in the development of airway infl ammation. Elevated ATP concentration in airway samples was reported in infl ammatory diseases. Airway ATP level is under tight control by enzymatic degradation, which could also affect ATP concentration measured in ex vivo biological samples. Aim: To assess ATP degradation in bronchoalveolar lavage (BAL) fl uid. Methods: ATP was measured in BAL collected from 20 subjects (6 patients with interstitial lung diseases, 4 with asthma, 2 with COPD, 1-1 with extrinsic allergic alveolitis / Wegener granulomatosis / microaspiration / pleuritis / pulmonary hypertension / pneumonia and 2 healthy controls) within an hour after collection using luminometry. ATP degradation was assessed as the recovery of 1 M ATP after 30 minutes. Seven samples were also collected on chelating solutions (0.32% sodium citrate or 2 mM EDTA+2 mM EGTA). Data were analyzed with non-parametric tests (median /interquartile range/). Results: BAL fl uid ATP concentration was 18 nM /4-107 nM/, and recovery of added ATP was 36% /29-82%/. ATP concentration was higher in samples treated with citrate (530 nM /375-715 nM/) or EDTA+EGTA (420 nM /234-828 nM/) compared to untreated samples (186 nM /39-302 nM/; p<0.05). ATP degradation was inhibited by both citrate (recovery: 99% /50-100%/) and EDTA+EGTA (recovery: 80% /53-100%/) in comparison with no treatment (p<0.05). Conclusion: Extracellular ATP is degraded at a variable speed in BAL fl uid, which is effectively inhibited by chelating agents. The analysis of pre-treated BAL fl uid might more precisely refl ect airway ATP concentration. Oral Presentation Room 3.10 10:45 12:45 Abstract printing supported by . Visit Pfizer Vaccines at Stand Q.01printing supported by . Visit Pfizer Vaccines at Stand Q.01 53s SUNDAY, SEPTEMBER 8TH 2013 399 Profi ling the proteome of the lower airway respiratory tract lining fl uid in chronic obstructive pulmonary disease Elif Melis Bicer1, Ben Forbes2, Graham Somers3, Anders Blomberg4, Annelie Behndig4, Ian Mudway1. 1Analytical and Environmental Sciences, King’s College London, London, United Kingdom; 2Insitution of Pharmaceutical Sciences, King’s College London, London, United Kingdom; 3Respiratory CEDD, GSK, Stevenage, United Kingdom; 4Public Health and Clinical Medicine, Lung Medicine, Umea, Sweden Chronic Obstructive Pulmonary Disease (COPD) is a chronic infl ammatory condition of the airways associated with protease, anti-protease imbalance and oxidative stress. Relatively little is known about how underlying airway immunopathology impacts upon the protective function of the respiratory tract lining fl uid (RTLF). Aim: To investigate compositional differences in the RTLF proteome in COPD patients compared against aged and smoking history matched controls. Broncholaveolar lavage samples were obtained from COPD smokers (n=5, 63.8±6.0 years), COPD ex-smokers (n=10, 66.0±6.8 years), healthy smokers (n=5, 61.4±6.2 years) and healthy non smokers (n=5, 66.8±5.9 years). Samples were analysed by one-dimensional gel electrophoresis and nanoliquid chromatography-tandem mass spectrometry, data processed using MASCOT and SCAFFOLD. We identifi ed 342 unique proteins, the greatest number, 153, observed in healthy non smokers, with only 49 proteins identifi ed in COPD ex-smokers. Proteins were classifi ed according to their gene ontology annotation, with COPD smokers found to possess the greatest number with roles in infl ammation, immune response and protease, anti-protease balance. Employing the sum of major ion intensities permitted a semi-quantitative assessment of protein concentrations with Calgranulin A, Surfactant protein A and Alpha-1 antitrypsin decreased in COPD smokers versus non smokers. Secretoglobulin concentrations were elevated in COPD patients versus smoking and non-smoking controls. Data suggests a simplifi ed RTLF proteome in COPD, with a shift in protein expression in COPD smokers consistent with ongoing infl ammation and protease, anti-protease imbalance. Oral Presentation Room 3.10 10:45 12:45 Abstract printing supported by . Visit Pfizer Vaccines at Stand Q.01printing supported by . Visit Pfizer Vaccines at Stand Q.01