about its suitability for the GFD. Previously, we analysed 15 quinoa cultivars using murinemonoclonal antibodies raised against both coeliac toxic gliadin and glutenins, we found that4 cultivars had small quantities of proteins with similar structure to some known toxicgluten epitopes. Therefore, we extracted prolamins from these 4 cultivars to investigate thereaction against gluten specific T-cells from intestinal biopsies. We measured proliferationof T-cells after prolamin stimulation using 3H-thymidine incorporation and the ability oftissue transglutaminase (TG2) to enhance T-cell recognition of these prolamins. Our resultssuggest that similar T-cell antigens exist within wheat prolamins and one quinoa cultivar.It is clear that TG2 deamidation can enhance this reactivity. However, the significance ofthese findings with respect to quinoa requires further evaluation.

S2047

Frequency of Elevated ALT in Untreated Coeliac Disease and the Impact ofCompliance With Gluten Free Diet on ALT NormalisationAnna Foley, Sue J. Shepherd, Peter R. Gibson

Elevated transaminases are often observed in patients with newly diagnosed coeliac disease(CD) with up to 57% being reported in selected populations. Their clinical significance isuncertain, although they are believed to be part of the coeliac inflammatory process andnormalise on a gluten-free diet (GFD). Aims: To determine the prevalence and factorsassociated with elevated ALT in patients with untreated, and the effect of GFD on ALT.Methods: 99 consecutive untreated patients with biopsy-proven CD referred for dietaryeducation in GFD were prospectively studied with regard to demographics, symptoms, LFTs,body composition and serology. The first 53 patients were followed regularly for 12 months,including LFTs. All were compliant with the GFD. These were compared to 60 age and sexmatched patients on a long term GFD (>2 years) who were in serological remission. Results:Mean age (range) was 39 (range 18-71) years. 24% were men. ALT was elevated (female>30 IU/L, male >40IU/ml) at diagnosis in 27% in whom ALT was 31 to 99. After > 2 yearsGFD 25% had persistently elevated ALT. Over 12m GFD 24% developed high ALT despitebeing normal at diagnosis. Abnormal ALT was not associated with symptom severity, Marshclassification, sex, HLA-D genotype or alcohol intake. There was a significant correlationbetween WHR and high ALT levels (r value 0.59, p=0.002), however no other body composi-tion data had a positive correlation. Conclusion: Elevated ALT is common (27%) at diagnosisin this unselected population. Contrary to popular belief, one third of these do not normaliseover 12 months with compliance to GFD and ALT can become chronically abnormal aftercommencing the GFD. The positive correlation with WHR (reflecting visceral fat) and ALTmay contribute to high cytokine load causing hepatic inflammation or may reflect hepaticsteatosis. The clinical significance of such “unstable” ALT levels requires elucidation.

S2048

Cell Imaging of a Fluorescently Labelled Gliadin Peptide Across LiveIntestinal Epithelial Monolayers Challenged With IFN-γAnn Chua, Guy R. Sander

Background: Increased translocation of intact gluten peptides across the intestinal epitheliumcontributes to the pathogenesis of coeliac disease. Largely unknown is how these peptidescross the epithelium and the effect of permeability increasing proinflammatory cytokinessuch as interferon-γ (IFN-γ). We describe the use of a novel technique using confocalmicroscopy to track a fluorescently labelled α-gliadin peptide (p57-89) across live intestinalepithelial monolayers grown on Transwells. We aimed to use the technique to determinethe cellular uptake of p57-89 in live intestinal epithelial monolayers during their challengewith IFN-γ, a key cytokine up-regulated in coeliac disease. Methods: HT-29 and T84 cellmonolayers were grown on semi-permeable plastic Transwell membranes. T84 monolayerswere challenged with IFN-γ (100ng/mL) for 48 hours. Peptide influx and ionic permeabilityas measured by transepithelial electrical resistance (TER) were analysed. Following cytokinechallenge of monolayers, FAM labelled p57-89 (FAM-p57-89; 0.2ng/ml) was added to theapical chamber of Transwells and incubated for a further 4 hours. Transwells were thenplaced on top of microscope slide cover slips and the location of FAM-p57-89 in liveintestinal epithelial monolayers directly visualised with a confocal microscope. Results:Following 48hrs of IFN-γ challenge of T84 monolayers, TER decreased 77% (923 +/- 52Ω/cm2 vs 213 +/- 19 Ω/cm2; p<0.01) and the rate of peptide influx increased 4.27 fold(+/- 0.1; p<0.01) compared to the unchallenged monolayers. Using confocal microscopy,we detected FAM-p57-89 intracellularly and along the lower half of the lateral membranewhen live T84 monolayers were challenged with IFN-γ but not in control monolayers. FAM-p57-89 was also detected in cytoplasmic vesicles and along the lower half of the lateralmembrane in the leakier HT-29 cell monolayer. Conclusions: The confocal microscope canbe used to detect and track fluorescently labelled gliadin peptides across live intestinalepithelial cell monolayers grown on Transwells. Cellular uptake of gliadin peptides duringIFN-γ challenge appears to be transcellular with peptide accumulating along the lateral mem-brane.

S2049

Celiac Disease Rate Increases in Patients With Irritable Bowel Syndrome- ACommunity Screen From West VirginiaYoram Elitsur, Joe E. Khoury, Karen Daniel

Introduction: Irritable bowel syndrome (IBS) and celiac disease (CD) may share similarclinical symptoms including abdominal pain and diarrhea. Patients with IBS are consideredas a high risk group for the development of CD. Indeed, previous reports have suggestedan increased CD rate in IBS patients (OR-7.0; Lancet 2001). Aim: We investigated the rateof CD in a cohort of adult patients diagnosed with IBS. Methods: A community health fair,organized by the Gluten Intolerance Group (GIG) of West Virginia, was performed inautumns of 2008 and 2009. The study included adults and children who have the followingconditions: IBS, autoimmune diseases, or positive family history of celiac disease. Anti-tissuetransglutaminase (tTG) antibody was utilized to establish the diagnosis of CD. The diagnosis

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of patients with IBS was confirmed by Rome III criteria. Results: One hundred and ninetythree subjects were screened, of whom 91 (47%) had IBS confirmed by Rome III criteria.The clinical characteristic was distributed as followed: IBS-D in 43, IBS-C in 31, IBS-M in16, and unsubtyped IBS in 1 patient. Only 2 (2.2%) patients with IBS had elevated tTGlevels. Small intestinal biopsies were performed in both subjects and were positive for celiacdisease. Conclusion: The diagnosis of celiac disease in our cohort of IBS adults is higherthan expected in the general population (approximately 1%). Celiac disease screening shouldbe considered in patients with IBS.

S2050

Gluten Sensitivity: Beyond Celiac DiseaseRaffaella Tortora, Paola Iovino, Ilaria Russo, Fabiana Zingone, Pietro Capone, AntonellaRanaudo, Paolo Andreozzi, Carolina Ciacci

Gluten sensitivity is a new concept to define a condition of some morphological, immunolo-gical and functional disorders that withdraw with a gluten free diet in patients HLADQ2/DQ8 positive, with negative gluten-specific autoantibodies and without histological character-istic of celiac disease (CD). Aim of this study was to characterize in the patients with glutensensitivity the mucosal responses during an In Vitro gluten challenge (iVGC) with expositionto gliadinin comparison with celiac patients on gluten-free diet. Methods: 50 CD patientson gluten-free diet and 59 patients reporting gluten sensitivity and negative specific antibodiesunderwent upper endoscopy and multiple jejunal biopsies. Mucosa was placed on organculture dish for 3 and 24 hours, challenged with peptic-tryptic digest from wheat which,in CD mucosa, is known to induce expression of markers of immunological activation (CD3,CD25, CD69, PY99). In fact, in all CD patients iVGC was able to show mucosal inflammationupon gliadin challenge. Basal histology showed normal mucosa in 12 gluten sensitive patients,increased IELs and mucosal alteration in 47 (Marsh 1: 36 patients; Marsh 2: 7 patients;Marsh 3: 4 patients). In 10 gluten sensitive patients iVGC showed mild mucosa inflammation(modest CD3/CD25 increase, CD69 and PY99 increase) as for some CD patients, in 47mucosa showed no signs of inflammation upon gliadin challenge, and in 2 iVGC was notconclusive because inducing a mild increase of some markers but not all. In conclusion: in79.6 % of patients reporting gastrointestinal symptoms after gluten ingestion iVGC was notable to disclose mucosal inflammation after gliadin challenge. In 16.9%, however, iVGCdisclosed a mild inflammation following gliadin contact. Our data for the first time demon-strate for a small percentage of gluten sensitive patients an immunological activation ofintestinal mucosa secondary to gliadin exposure.

S2051

Celiac Disease and Risk of Colorectal NeoplasiaRaquel Gonzalez, Lisandro Pereyra, Adriana Mohaidle, José M. Mella, Carolina Fischer,Mario A. Medrano, Beatriz Vizcaino, Adrián R. Hadad, Pablo Luna, Daniel G. Cimmino,Silvia C. Pedreira, Luis A. Boerr

Introduction: although small bowel and esophagus neoplasia are recognized to occur morefrequently in patients with celiac disease (CD), an association with colorectal cancer has notbeen described. Aim: To determine the risk of colorectal neoplasia among patients withceliac disease. Materials and methods: A nested case-control study was conducted using theGastroenterology and Endoscopy electronic data base to identify patients that had performedcolonoscopy. Patients with CD were regarded as “cases” and those without CD were regardedas “controls”. For each case, two controls matched for age, sex, colonoscopy purpose andfirst and second grade family history of colorectal cancer, were randomly selected. A surveywas carried out by telephone calls to assess patients on their risk factors and history disease.The main outcome evaluated was the risk of colorectal polyps, adenomas, advanced lesionsand cancer. Risk was measured in odds ratio (OR) and its corresponding confidence intervals95% (CI). Results: Out of 178 celiac disease patients analyzed, 44 had undergone a previouscolonoscopy and were included in the study as cases and 88 as controls. In cases, the averageage was 55 years old (range 34-82 years old), 86% women, the colonoscopy purpose wascolorectal cancer screening in 63% and abdominal pain in 16% and strict adherence to thegluten- free diet occurred in 66%. Presence of polyps, adenomas and advanced coloniclesions was 9/44 (20%), 7/44 (16%) and 2/44 (4, 5%) respectively. In controls, the averageage was 55 years old (range 33-82 years old), women 86%, and the colonoscopy purposewas colorectal cancer screening in 55% and abdominal pain in 15%. Presence of polyps,adenomas and advanced colonic lesions was 13/88 (15%), 8/88 (9%) and 3/88 (3, 4%)respectively. The risk of polyps, adenomas and advanced colonic lesions was similar in bothgroups (OR 1, 48, CI 0, 59- 3, 73 p = 0, 40, OR 1, 89, CI 0, 66- 5, 42 p= 0, 24, and OR1, 34 CI 0, 26-7, 05 p= 0, 74, respectively). No colorectal cancer was identified. Conclusion:The risk of colorectal neoplasia within a cohort of patients with celiac disease, who mostlypresented a strict adherence to gluten- free diet, was similar to the general population.

S2052

Relevance of the Manual Orientation of the Endoscopic Biopsies in theDiagnosis of Gluten-Sensitive EnteropathyFrancisco J. Martínez-Cerezo, Gemma Castillejo, Vanessa Morente, Joan Marsal, FranciscoJ. Tena, Vicente Moreno, Francisco Riu, Domingo Pascual

The method of choice for the histopathological diagnosis of gluten-sensitive enteropathy isthe obtention of multiple duodenal endoscopic biopsies. It is important for the biopsies tobe of good quality for its correct interpretation. The aim was to assess the proportion ofendoscopic duodenal biopsies well oriented and to compare the quality of biopsies with orwithout its manual orientation. Methods. Endoscopic duodenal biopsies obtained frompatients with suspected gluten-sensitive enteropathy were included. Both adult and pediatricpatients were recruited; informed consent was obtained from all patients or their guardians.Antitransglutaminase antibodies and haplotypes HLA DQ2 and DQ8 were determined previ-ously. At least four biopsies were obtained for all patients; half of them were manuallyorientated and stretched with the naked-eye over a vinyl surface with forceps before their

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