Astrid Konrad

Resistin is an inflammatory marker of inflammatory bowel disease in humans.

Objectives Resistin, a recently discovered adipokine, has been shown to be associated with inflammatory conditions such as insulin resistance, obesity, atherosclerosis and rheumatoid arthritis. We therefore hypothesized that (i) resistin levels may be elevated in patients with inflammatory bowel disease (IBD) and (ii) resistin levels may be associated with disease activity in IBD. Methods We addressed these questions by testing for associations between resistin plasma levels, inflammatory parameters and clinical disease activity in a case–control study with 235 patients with Crohns disease (CD), 112 patients with ulcerative colitis (UC) and 144 healthy controls. Results Patients with IBD showed significantly higher resistin levels compared with controls (P<0.0001). In both, patients with CD and UC, resistin concentrations were significantly associated with elevated white blood cell count (P<0.0001), C-reactive protein (CRP) (P<0.0001) and disease activity (P≤0.0001). In multivariate logistic regression analysis, resistin levels were identified as an independent predictor of active disease (odds ratio 1.014, 95% confidence interval 1.002–1.027, P=0.02) in patients with CD after adjusting for sex, age, body mass index, white blood cell count and CRP. In UC patients, resistin was associated with active disease in multivariate regression analysis after control for sex, age, body mass index and white blood cell count (odds ratio 1.015, 95% confidence interval 1.002–1.029, P=0.02). Addition of CRP, however, abolished this association. Conclusion Resistin levels are an independent predictor of disease activity in patients with CD. Resistin may represent a novel link between inflammation and IBD.

Curcumin induces the tolerogenic dendritic cell that promotes differentiation of intestine-protective regulatory T cells.

The gut is home to a large number of Treg, with both CD4+ CD25+ Treg and bacterial antigen‐specific Tr1 cells present in normal mouse intestinal lamina propria. It has been shown recently that intestinal mucosal DC are able to induce Foxp3+ Treg through production of TGF‐β plus retinoic acid (RA). However, the factors instructing DC toward this mucosal phenotype are currently unknown. Curcumin has been shown to possess a number of biologic activities including the inhibition of NF‐κB signaling. We asked whether curcumin could modulate DC to be tolerogenic whose function could mimic mucosal DC. We report here that curcumin modulated BM‐derived DC to express ALDH1a and IL‐10. These curcumin‐treated DC induced differentiation of naïve CD4+ T cells into Treg resembling Treg in the intestine, including both CD4+CD25+ Foxp3+ Treg and IL‐10‐producing Tr1 cells. Such Treg induction required IL‐10, TGF‐β and retinoic acid produced by curcumin‐modulated DC. Cell contact as well as IL‐10 and TGF‐β production were involved in the function of such induced Treg. More importantly, these Treg inhibited antigen‐specific T‐cell activation in vitro and inhibited colitis due to antigen‐specific pathogenic T cells in vivo.

Generation of Antigen-Specific, Foxp3-Expressing CD4+ Regulatory T Cells by Inhibition of APC Proteosome Function

We tested the hypothesis that immature APC, whose NF-κB-signaling pathway and thus maturation was blocked by the proteosome inhibitor benzyloxycarbonyl-isoleucyl-glutamyl(O-tert-butyl)-alanyl-leucinal (PSI), could be a source of Ag-specific regulatory T (Treg) cells. DO11.10 CD4+ T cells that were incubated with Ag- and PSI-pulsed APC proliferated poorly, produced less IL-2, IFN-γ, and IL-10 in secondary cultures, and inhibited the response of both naive and memory CD4+ T cells stimulated by Ag-pulsed APC. The generation of PSI-APC Treg cells required IL-10 production by APC. PSI-APC Treg cell inhibition required cell-cell contact but not IL-10 or TGF-β. Addition of IL-2 did not reverse, but Ab to CTLA-4 did reverse partially the inhibitory effect. Depletion of CD25+ T cells before initial culture with PSI-APC did not affect Treg generation. PSI-APC Treg cells expressed high levels of Foxp3, inhibited proliferation of naive DO11.10 T cells in vivo, and abrogated colitis driven by a memory Th1 response to bacterial-associated Ag. We conclude that NF-κB-blocked, immature APC are able to induce the differentiation of Treg cells that can function in vitro and in vivo in an Ag-specific manner.

Probiotics and Immune Regulation of Inflammatory Bowel Diseases

Intestinal microflora play an important role in the pathogenesis of inflammatory bowel diseases (IBD). Certain probiotic bacteria provide beneficial effects for human health and intervention of IBD. Possible mechanisms underlying these effects are diverse and include many aspects of the interaction of the host with its commensal microflora, with immunological and non-immunological effects. This review paper will focus on the recent progress in the use of probiotics in the treatment of IBD, and discuss the potential immunological mechanisms underlying their effects, such as the modulation of mucosal T cell, B cell, epithelial cell, dendrtic cell, macrophage, nature killer cell, antibody, and cytokine responses.

Pharmacogenetics of Crohn's disease

The considerable interindividual differences in efficacy and side effects of commonly used medications in Crohns disease are partly owing to genetic polymorphisms. Many genetic variants have been studied in genes possibly involved in the metabolism or mechanism of action of therapeutic agents such as glucocorticosteroids, azathioprine/6-mercaptopurine, methotrexate, calcineurin inhibitors or anti-TNF agents. However, the only test translated into clinical practice is thiopurine S-methyltransferase (TPMT) genotyping for hematological toxicity of thiopurine treatment. To date, there are no other meaningful applications for pharmacogenomics in clinical practice of Crohns disease. In the future, designed therapeutic trials should possibly permit the development of predictive models including genotypic markers, such as that proposed for the clinical outcome after infliximab therapy, which includes an apoptotic pharmacogenetic index. The recent identification of new susceptibility genes provides additional candidate markers that have possible effects on the outcomes of therapies, and prioritizes new therapeutic targets, such as the IL-23 pathway. Further innovative approaches might be relevant for the pharmacogenetic investigation of gene variants implied in innate immune pattern recognition and autophagy.

Gene disruption and immunity in experimental colitis.

In recent years, a number of experimental models of chronic intestinal inflammation have been generated that provide insights into the complex mechanisms that can result in chronic intestinal inflammation. Most of the models in some way affect T cell function and these can be further subdivided into two categories, based on the concept of disease resulting from an imbalance between regulatory and effector T cells. Thus, the one category represents instances where impaired regulatory T cell activity appears responsible for disease; the second represents models in which excessive effector T cell function seems to drive disease. A third category includes models in which perturbations of the epithelium appear to result in chronic intestinal inflammation. Last, we will discuss a spontaneous model of disease, the C3H/HeJBir mouse, which involves a strong genetic predisposition. These and other models have been recently reviewed in some detail.

M2042 The Role of Pregnane X Receptor (PXR/NR1i2) Gene Variants in Inflammatory Bowel Disease

s of the 4th Congress of ECCO the European Crohn’s and Colitis Organisation S111 colitis (UC) 2 3 cases per 100.000 per year. So far the incidence of pediatric IBD in Hungary is unknown. Methods: The participating institutes are requested to fill out a questionnaire (78 parameters) about every newly diagnosed IBD patients younger than 18 years. The questionnaire is about epidemiological and antropometrical data, main symptoms, diagnostic procedures (endoscopy, CT, MRI), and the detailed results of histological and imaging procedures. Results: Between 01.01.2007 and 30.09.2008 223 newly diagnosed cases of IBD were prospectively identified: 139 cases of CD, 69 cases of UC and 15 cases of indeterminate colitis. As a result the incidence of childhood IBD was 6.13 cases per 100.000, with the incidence of CD being 4.07 cases per 100.000 per year, the incidence of UC 1.67 cases per 100.000 per year and the incidence of indeterminate colitis 0.40 cases per 100.000 per year. The mean age at diagnosis was 13 years (range: 1.5 18 year). There was a male preponderance in CD, in contrast, sex ratio in UC patients were equal. Positive family history of IBD was registered in 7.8% of patients and 9.2% of patients with CD were reported to have a fistula. Ileoscopy rate was only 48% technical problem was the most common reason for the lack of ileal intubation. Oesophagogastro-duodenoscopy was performed in 50% of all cases. In 35% were MRI or CT scan made for the detailed verification of the disease. Conclusions: The incidences reported in the first 21 months of HUPIR are similar to the European and North American data. The dominance of CD proved to be also consistent with other studies. Almost 10% of the patients with CD had fistula. Ileal intubation and oesophago-gastro-duodenoscopy were performed in the half of the cases, and this rate should be improved in the future. P253 The role of pregnane X receptor (PXR/NR1I2) gene variants in inflammatory bowel disease J. Seiderer1 *, J. Glas1, J. Diegelmann1, D. Fischer1, B. Seitz1, S. Pfennig1, T. Griga2, W. Klein3, J.T. Epplen3, U. Schiemann4, T. Mussack5, B. Goke1, T. Ochsenkuhn1, M. Folwaczny6, B. Muller-Myhsok7, S. Brand1. 1University of Munich Klinikum Groshadern, Munich, Germany, 2Department of Internal Medicine, Knappschaftskrankenhaus Dortmund, Dortmund, Germany, 3Department of Human Genetics, Ruhr-University, Bochum, Bochum, Germany, 4Department of General Internal Medicine, Inselspital Bern, Bern, Switzerland, 5University of Munich Department of Surgery, Munich, Germany, 6Clinic for Preventive Dentistry and Parodontology, LMU Munich, Munich, Germany, 7Biostatistics, Max-Planck-Institute for Psychiatry, Munich-Schwabing, Munich, Germany Introduction: The pregnane X receptor (PXR), also known as NR1I2 (nuclear receptor subfamily 1, group I, member 2), is a nuclear receptor encoded by the PXR/NR1I2 gene. The PXR/NR1I2 gene has recently been reported to be associated with susceptibility to inflammatory bowel disease (IBD) (Gastroenterology 2006;130:341 8). However, since a subsequent case-control study failed to replicate this association in an independent population, further replication studies in different ethnic cohorts are required. We therefore investigated this potential association in a large European IBD cohort. Aims and Methods: Genomic DNA from 2823 Caucasian individuals including 859 patients with Crohn’s disease (CD), 464 patients with ulcerative colitis (UC), and 1500 healthy unrelated controls was analyzed for eight single nucleotide polymorphisms (SNPs) in the pregnane X receptor (PXR/NR1I2) gene (rs12721602, rs3814055, rs1523128, rs1523127, rs12721607, rs6785049, rs2276707, rs3814057). Genotyping was performed by PCR and melting curve analysis using a pair of fluorescence resonance energy transfer (FRET) probes in a Light Cycler. Results: With the exception of a weak association of rs2276707 with UC (p = 1.02 x 10 2; OR 1.27 [1.06 1.52]), none of the analyzed variants in the pregnane X receptor (PXR/NR1I2) was associated with susceptibility to CD or to UC in our study population. Conclusion: Our data could not confirm the association of PXR gene variants with CD or UC. Further studies investigating the phenotypic effect and potential epistatic interactions of PXR with other IBD susceptibility genes are required to determine the exact role of PXR in IBD. P254 Incidence of nonspecific ulcerative colitis in Republic of Tatarstan (first Russian inflammatory bowel diseases registry) R.A. Abdulkhakov, S.R. Abdulkhakov*, A.A. Abbakumova. Kazan State Medical University, Kazan, Russian Federation The purpose of the study was to find out the real incidence of nonspecific ulcerative colitis (NUC) in Republic of Tatarstan. Materials and Methods: The registry of NUC patients was made according to case histories and out-patient charts including those who presented with the first onset of the disease as well as those with relapses or stable remission. Results: According to the registry there are 428 NUC patients in Republic of Tatarstan (m-216, 50.47%, f-212, 49.53%, NS). Incidence of NUC is 11.4 per 100000 of population. Acute NUC was revealed in 13 (3%) patients, chronic persistent NUC in 38 (8.9%) patients. Most of the patients (277 pts, 64.7%) had chronic relapsing type of disease which was observed quite often both in males (138 pts) and females (139 pts). In case of 76 patients it was first admission to the hospital due to complaints associated with NUC. In most of patients (150 pts, 35%) distal colitis was observed, in 89 (21%) total colitis, in 32 (7.5%) subtotal, left-sided colitis was found in 125 (29.2%) patients, in case of 32 patients there were no data about the localization of inflammation. According to severity of inflammation distribution of patients was the following: mild inflammation was found in 70 (16.4%) cases, moderate in 237 (55.4%), severe in 61 (14.3%) patient, in case of 60 patient data was lacking. Massive bleeding as a complication of NUC occurred in 10 (2.3%) patients, 4 of them underwent surgery. NUC was the cause of disability in 79 (18.5%) patients. Conclusions: It’s the first registry of NUC patients in Republic of Tatarstan which covered 428 NUC patients and revealed incidence of NUC as 11.4 per 100000 of population. NUC incidence is the same in men and women. Most of patients have distal type of NUC with moderate activity. Frequency of massive bleeding as NUC complication is 2.3%. P255 Anxiety and depression symptoms in Crohn’s disease patients in remission M. Iglesias1, M. Barreiro2 *, A. Figueiras3, M. Vazquez4, L. Nieto1, M. Seoane2, A. Lorenzo2, J.E. Dominguez-Munoz2. 1FIENAD University Hospital, Santiago de Compostela, Spain, 2Gastroenterology University Hospital, Santiago de Compostela, Spain, 3Epidemiology University Hospital, Santiago de Compostela, Spain, 4Clinic Psycology University of Santiago, Santiago de Compostela, Spain Background: Crohn’s disease (CD) is a chronic inflammatory bowel disease with periods of relapse and remission. Role of anxiety and depression in CD patients in remission has been poorly studied. We hypothesized that despite staying in remission, anxiety and depression symptoms have an import role in CD patients. Aim of the study was to evaluate the presence of anxiety and depression symptoms in CD patients in remission and potential by gest on A uust 4, 2016 http://eccoxfordjournals.org/ D ow nladed from

M2041 The First Two Crohn's Disease Susceptibility Loci with a High Degree of Epistasis: PTGER4-Expression-Modulating Polymorphisms in the 5p13.1 Region Enhance ATG16L1-Associated Susceptibility to Crohn's Disease

J. Seiderer1, J. Glas1, J. Diegelmann1, G. Pasciuto1, C. Tillack1, D. Roeske2, S. Pfennig1, M. Jurgens1, A. Konrad1, H. Torok1, T. Griga3, W. Klein4, J.T. Epplen5, U. Schiemann6, T. Mussack7, P. Lohse8, B. Goke1, T. Ochsenkuhn1, M. Folwaczny9, B. Muller-Myhsok10, S. Brand1 *. 1University of Munich Klinikum Groshadern, Munich, Germany, 2Biostatistics, Max-Planck-Institute for Psychiatry, Munich-Schwabing, Munich, Germany, 3Department of Internal Medicine, Knappschaftskrankenhaus Dortmund, Dortmund, Germany, 4Department of Human Genetics, Ruhr-University, Bochum, Germany, 5Department of Human Genetics, University Bochum, Bochum, Germany, 6Department of General Internal Medicine, Inselspital Bern, Bern, Switzerland, 7Department of Surgery, LMU Munich, Munich, Germany, 8Department of Clinical Chemistry, University Hospital Munich-Grosshadern, Munich, Germany, 9Clinic for Preventive Dentistry and Parodontology, LMU Munich, Munich, Germany, 10Max-Planck-Institute for Psychiatry, Munich-Schwabing, Munich, Germany

13-hydroxy-octadecatrienic acid (Hox alpha) ameliorates chronic murine colitis

Introduction: ~he therapeutlc options for inflammatory bowel disease (IBD) remain limited, especially tor remission prolongation knti-TNF-a antibodies such as lntltxinmb are effective in the treatment ol flares but ohen lose their ethcacy with repetitive applications, 13Hydroxyoctadecatnen*~ acid a componem of the sungnig nettle leaf (Hox alpha, Strathmann (krrnany) reduces TNF-c~ secretion and is a known therapy m the setting of rheumatoid arthritis Aim: 1o investigate the etlect of Hox alpha on murine colitis disease activity in difli:rent colitic mouse models Methods: C3}tIHeJBir,IL-10-/and BALB/{ mice with DSSreduced acute and cbronic colitis were treated with 0 5mg/ml Hox alpha and compared to untreated controls Mice were clniically monitorvd and inflammation was assessed by histological scoring, analysis oi kcal/I.113 and measurement ot mucosal cytokine production by EL/SA Probkration of lynlphocytes ot the spleen and Peyers patches was quantified by 3H-thynlidine nico~)oration. Resuhs: Hox alpha-treated mice ,aqth chronic DSS colitis as well as Hox alpha-treated ID 10-A mice displayed signihcantly reduced clniical and histologB <al signs ot colitis than untreated mice (p<0 05) Fecal IL-113, mucosal IL-113 as well as TNF-~ concentrations were signit~cantly lowe* in Hox alpha-treated chronic DSS mice as well as in ttox alpha-treated lL-10-/mice m comparison with control rnice (p<0.05). Furthe*r~mre, 1ympbocyte prolfl~ration postdipopolysaccharide stimulation was significantly reduced in Hox alpha-treated mice ( p < 0 0 0 1 ) Conclusions: The continuous use of Hox alpha is effective in the amelioration of chronic murme colitis, an effect we attribute to a decreased Thl immune response Hox alpha thus possibly presents us with a new therapeutic option fur prolonging remission in 1BD.