SUNDAY,SEPTEMBER 25 TH 2011 64. Cystic fibrosis: new aspects of diagnosis, inflammation and detecting exacerbation 361 Intestinal current measurement (ICM) as a new diagnostic test for cystic fibrosis (CF) Malena Cohen-Cymberknoh 1,2 , Yasmin Yaakov 1,3 , Eitan Kerem 1,2 , David Shoseyov 1,2 , Joseph Rivlin 4 , Lea Bentur 5 , Elie Picard 6 , Micha Aviram 7 , Michael Wilschanski 1,3 . 1 CF Center, 2 Department of Pediatrics, 3 Department of Pediatric Gastroenterology, Hadassah-Hebrew University Medical Center, Jerusalem, Israel; 4 CF Center, Carmel Medical Center, Haifa, Israel; 5 CF Center, Rambam Medical Center, Haifa, Israel; 6 CF Center, Shaare Zedek Medical Center, Jerusalem, Israel; 7 CF Center, Soroka Medical Center, Beer Sheva, Israel Background: Like the nasal potential difference (NPD) test, ICM may be useful for the diagnosis of atypical CF. However, ICM is easily applicable at all ages. Aim: To assess the diagnostic reliability of ICM in a large cohort of CF, healthy control and patients with questionable CF. Methods: Rectal biopsies were taken from 3 groups: known CF patients, healthy controls and patients with questionable CF. The last group had a variety of symp- toms suggestive of CF: recurrent pneumonia, unexplained bronchiectasis, chronic diarrhea and/or failure to thrive. ICMs were performed using standard protocols by mounting the rectal biopsy in an Ussing chamber and sequentially adding secretagogues while recording current changes. Results: 17 known CF patients and 16 control patients were examined and have remarkably different results (all results are presented as μA/cm 2 ): carbachol 16±7, histamine 13±9 and forskolin 4.8±4 for healthy control group and carba- chol -3.7±6.8 (p<0.0001) histamine -3.1±2.7 (p<0.0001) and forskolin 0.2±0.4 (p=0.0004) for the CF group. The suggested reference values are: +3.75, +0.25, +1.32 for carbachol, histamine and forskolin, respectively. The combination score (the sum of the 3 secretagogues) differentiates normal from abnormal ICM (ROC Curve analysis, area under the curve =1.00, both sensitivity and specificity are 100%). This statistical model was applied to 70 patients suspected for CF and revealed that 59 patients had normal and 11 patients had abnormal ICM results. Conclusion: In this study we have shown that ICM tests may be useful to dif- ferentiate between CF and non-CF patients and may be included in diagnostic algorithms. Larger studies are needed to confirm these results. 362 Regulation of ion transporters and airway surface dynamics by lipoxin in cystic fibrosis bronchial epithelium Mazen Al-Alawi 1 , Valia Verriere 1 , Richard Chostello 2 , Valerie Urbach 1 , Brian Harvey 1 . 1 Department of Molecular Medicine, Royal College of Surgeons in Ireland, Dublin, Ireland; 2 Department of Respiratory Medicine, Royal College of Surgeons in Ireland, Dublin, Ireland Aims: We have investigated the role of the endogenous anti-inflammatory lipoxin LXA 4 in modulating Cl - secretion and Na + absorption, airway surface liquid height (ASLh) and ciliary beat frequency (CBF) in CF and non-CF bronchial epithelia. Methods: CF (CuFi-1) and non-CF (NuLi-1) bronchial epithelial cell lines were grown under an air-surface liquid interface into well-differentiated epithelia. ASLh and CBF were measured using confocal fluorescence microscopy and ion transport using patch-clamp and short-circuit current techniques. Results: LXA 4 (1nM) treatment for 15 minutes, increased ASLh by 47.5±0.5% and 103.0±3.0% in NuLi and CuFi epithelia respectively (P<0.001, n=18). The stimulatory effect of LXA 4 on ASLh was sustained over 24 hours in the CF ep- ithelia and was inhibited by the following pre-treatments: bumetanide, amiloride, Boc-2 (LXA 4 receptor antagonist), reactive blue (P2Y receptor antagonist) and extracellular hexokinase (ATP hydrolysis). LXA 4 stimulated CBF, intracellular Ca 2+ mobilization, Cl - secretion and inhibited Na + absorption in the CF epithelia. Conclusions: These effects of lipoxin involving the FPR2 receptor, apical ATP release, purinoreceptor activation, inhibition of Na + absorption and stimulation of Cl - secretion to enhance airway surface liquid dynamics open up a new therapeutic avenue to promote mucociliary clearance in cystic fibrosis airways. 363 Deficient production of IFN-stimulated genes upon rhinovirus infection in cystic fibrosis airway epithelial cells Elisabeth Kieninger 1 , Marjolaine Vareille 1 , Marco P. Alves 1 , Brigitte S. Kopf 1 , Thomas Geiser 2 , Sebastian L. Johnston 3 , Michael R. Edwards 3 , Nicolas Regamey 1 . 1 Division of Paediatric Respiratory Medicine, University Children’s Hospital, Bern, Switzerland; 2 Division of Pulmonary Diseases, University Hospital of Bern, Bern, Switzerland; 3 Department of Respiratory Medicine, National Heart & Lung Institute, Imperial College London, London, United Kingdom Background: Rhinoviruses (RVs) are important triggers of pulmonary exacerba- tions and possible contributors to long-term respiratory morbidity in cystic fibrosis (CF), but mechanisms leading to RV-induced CF exacerbations are poorly under- stood. We recently described deficient innate immune responses to RV infection in CF characterized by impaired type I and III interferon (IFN) production and increased virus replication. To study downstream effects of impaired IFN induction we investigated the ex- pression of IFN-stimulated genes (ISGs) which are important for the production of antiviral proteins. Methods: Epithelial CF and non-CF cell lines (UNCCF2T/UNCN2T, CFBE41o- /16HBE14o-) were cultured and infected with RV-16 and -1B at a MOI of 2. Induction of ISGs including MxA, 2’,5’-OAS, viperin and NOS2 was assessed by RT-PCR. Exogenous IFN-β and -λ were added before and after infection. Results: Expression of all ISGs was induced in CF and control cells upon virus infection. CF cells expressed 100-1000 times less ISGs than control cells (all p<0.05). ISG expression and RV replication were inversely related (MxA: r=- 0.79, p=0.001). There was a positive correlation between ISG expression and IFN-β (2’,5’-OAS: r=0.74, p=0.004) and IFN-λ production (NOS2: r=0.65, p=0.01). Ex- ogenous IFN increased levels of ISGs to the level of control cells, with a more pronounced effect of IFN-β. Conclusions: ISG induction upon RV infection is deficient in CF indicating a profound impairment of the early innate antiviral response. Addition of exogenous IFN restores antiviral pathways in CF, suggesting a potential use of IFNs in the prevention or treatment of RV-induced CF exacerbations. 364 Association of FCN1 and FCN2 gene polymorphisms with earlier onset of chronic pseudomonas aeruginosa (Pa) colonisation in cystic fibrosis (CF) patients Filomeen Haerynck 1 , Kristel Van Steen 2 , Tom Cattaert 2 , Bart Loeys 3 , Sabine Van Daele 1 , Petra Schelstraete 1 , Frans De Baets 1 . 1 Pediatric Pulmonology and Immunology, Ghent University Hospital, Ghent, Belgium; 2 Statistical Genetics, University of Liege, Liege, Belgium; 3 Medical Genetics, Antwerp University Hospital, Antwerp, Belgium Background: CF is a multisystem disease with high degree of phenotypic vari- ability especially in lung disease.Modifying genes of innate immunity may be involved in early onset of Pa colonisation. Methods: 82 Single Nucleotide Polymorphisms (SNPs) in 22 genes contributing to the innate immunity (MBL2, MASP (MBL associated serine Protease) 1/2/3, FCN (Ficolin) 1/2, LBP (Lipopolysaccharide-binding Protein), CD14,TLR (Toll- likereceptors 1→10)) were genotyped in a cohort of 116 CF patients. (age 6-44 years) Association survival analysis (Kaplan Meier and Cox regression) using additive, recessive, dominant and codominant model was performed looking for an association between SNPs and age of onset of Pa colonisation in all CF patients. Results: CF patients being heterozygous or homozygous for the mutant allel of both linked SNPs FCN1 (promoter) (A/G) and FCN1 (Q272Q) (exon 9) (G>A) are earlier colonised with Pa (p=0,016,p=0,026 resp). Earlier onset of Pa colonisation is seen in CF patients homozygous for mutant allel of -64A>C polymorphism FCN2 (promoter) (p=0,0031) and in patients having at least one mutant allel of the linked S258A (G>T)) polymorphism FCN2 (p=0,0057). CF patients heterozygous for mutant allel of TLR10 (rs7694115) (promoter) (T>C) (p=0,026) and linked SNP (rs11096957) (ex3) (N241H) (p=0,0113) and SNPs (rs11466645) (pro) (T>A) (p=0,0068), (rs11096956) (ex3) (P344P) (p=0,0067) are significantly later colonised with Pa. Conclusion: Mutant allel of SNPs FCN1 (pro and Q272Q) and FCN2 (-64A>C and S258A) is significantly associated with earlier Pa colonisation. Mutant allel of polymorphism of TLR10 is associated with later onset of Pa colonisation. 365 The polyamine spermine is increased in cystic fibrosis airway secretions Hartmut Grasemann 1 , Darakhshanda Shehnaz 1 , Paul Pencharz 2 , Felix Ratjen 1 . 1 Pediatrics, Respiratory Medicine, The Hospital for Sick Children, Toronto, ON, Canada; 2 Gastroenterology, Hepatology and Nutrition, The Hospital for Sick Children, Toronto, ON, Canada Rationale: Sputum arginase contributes to the nitric oxide (NO) deficiency in cystic fibrosis (CF) airways. Ornithine, the product of arginase activity, is the precursor of polyamines, which may play a role in the pulmonary response to injury and remodeling. Objective: To measure concentrations of spermine in sputum of CF patients. Methods: Using mass spectrometry, spermine was measured in sputum of clin- ically stable patients with CF (n=10), CF patients before and after antibiotic treatment for a pulmonary exacerbation (N=10) and healthy controls (n=10). CF patients were 7-17 years of age. Mean FEV1 in the stable CF patients was 80.4 (range 47-117)% of predicted values. FEV1 in CF patients presenting with a pulmonary exacerbation was 50.8±3.5 (range 36-69)% of predicted and improved by 13.5 (±2.8)% with treatment. Results: Mean (±SEM) spermine concentration in sputum was significantly higher in stable CF than controls (1.71±0.59 vs. 0.22±0.05 μmol/ml, p=0.02). Spermine concentrations were highest in CF patients presenting with a pulmonary exacerbation (9.10±1.62 μmol/ml) and decreased to levels similar to stable CF (1.68±0.33 μmol/ml, p<0.001, paired t-test), but remained significantly increased when compared to controls (p<0.001). The change in spermine concentrations during treatment for a pulmonary exacerbation correlated significantly with the 39s Oral Presentation Forum - 10:45-12:45 Abstract printing supported by . Visit Chiesi at Stand D.30