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P.62 INFLIXIMAB MODULATES MUCOSAL CYTOKINE PROFILE IN PATIENTS WITH INFLAMMATORY BOWEL DISEASES
Abstracts / Digestive and Liver Disease 42S (2010) S61–S192 was significantly reduced compared to non-inflamed areas, both in CD and UC patients, (CD: Median = 0.34, Q1-Q3 = 0.17-0.74, p=0.001; UC: Median = 0.47, Q1-Q3 = 0.24-0.77, p=0.0004), irrespective of the risk genotypes for the rs2631367 variant. The differences remained still significant when comparing the normalized quantities (CD: p=0.0001; UC: p=0.000056). Conclusions: The OCTN2 and IL23R mRNA levels are significantly reduced in inflamed colonic mucosa, in both CD and UC patients, irrespective of patients’ genotypes. # G. Inflammatory bowel diseases - 1. Basic science
P.62 INFLIXIMAB MODULATES MUCOSAL CYTOKINE PROFILE IN PATIENTS WITH INFLAMMATORY BOWEL DISEASES F. Caprioli ∗ ,1 , F. Bosè 2 , L. Raeli 2 , C. Viganò 3 , G. Basilisco 1 , D. Conte 3 , S. Abrignani 2 , E. Reali 2 1 Gastroenterology
Unit 2. Fondazione IRCCS Ospedale Maggiore Policlinico, Mangiagalli e Regina Elena, Milano; 2 INGM-National Institute of Molecular Genetics, Milano; 3 Department of Medical Sciences, Università degli Studi di Milano, Milano Background and aim: Monoclonal antibodies against tumor necrosis factoralpha (TNFα) are highly effective in the treatment of patients with inflammatory bowel diseases (IBD), even if their mechanism of action has yet to be fully elucidated. In patients with psoriasis, TNFα neutralization reduces dermal expression of proinflammatory cytokines produced by Th1 and Th17 lymphocytes, T cell lineages critically involved also in the pathogenesis of IBD. In this study, we aimed to evaluate the effect of TNFα neutralization by infliximab in modulating mucosal cytokine profile in patients with IBD. Material and methods: Twelve patients with active left-sided colonic IBD (6 patients with Crohn’s disease (CD) and 6 with ulcerative colitis, 7M, median age 34 yrs, range 25-50), and naïve to anti-TNF therapy were enrolled in the study. All patients underwent induction therapy with infliximab (5 mg/kg administered at week 0, 2 and 6). At enrolment and at week 6 a sigmoidoscopy was performed, and samples were obtained from the sigmoid colon. RNA was extracted from mucosal samples and quantitative expression of 96 inflammation-related genes was evaluated by qPCR, by means of TaqMan Low Density Array Human Immune Panel (Applied Biosystems). Mucosal gene expression was evaluated before and after therapy, and compared with results obtained from 16 controls with normal-appearing mucosa (6M, median age 38 yrs, range 20-44). Results: Among the panel of 96 inflammation-related genes, 36 were found to be significantly upregulated in the mucosa of patients with active IBD in comparison to controls. As compared to baseline, 6 weeks therapy with infliximab induced a significant downregulation of the expression of interleukin (IL)17A, IL-1β, IL-1α, IL-6, IL-8 and NOS2A (all P<0.05). In the CD patients subgroup, TNFα neutralization led to a reduced expression of interferongamma (IFNγ) and lymphotoxin-α, and to an upregulation of the expression of IL-4. With the exception of IL-8 and NOS2A, no significant differences were observed between the post-therapy levels of the above-mentioned genes and those measured in controls. Conclusions: Our data demonstrate that TNFα neutralization rapidly normalizes mucosal expression of several inflammation-related genes, including the signature Th1 and Th17-cytokines IFNγ and IL-17A. These results provide further insights on the mechanisms of anti-TNF agents in controlling mucosal inflammation. # G. Inflammatory bowel diseases - 1. Basic science
S125
P.63 INTERLEUKIN (IL)-17A BUT NOT IL-17E IS PROFIBROTIC IN CROHN’S DISEASE L. Rovedatti ∗ ,1 , A. Di Sabatino 1 , F. Vidali 1 , D. Fina 2 , P. Biancheri 1 , C. Ubezio 1 , A. Pasini 1 , G. Sampietro 3 , C. Alvisi 4 , M. Perego 4 , D. Foschi 3 , T. MacDonald 5 , G. Monteleone 2 , G. Corazza 1 1 First Department of Medicine, Fondazione IRCCS Policlinico S. Matteo, Centro per lo Studio e la Cura delle Malattie Infiammatorie Croniche Intestinali, University of Pavia, Pavia; 2 Department of Internal Medicine, University Tor Vergata, Roma; 3 Surgery Division, Department of Clinical Sciences, Luigi Sacco University Hospital, Milano; 4 Endoscopy Unit, Fondazione IRCCS Policlinico S. Matteo, Centro per lo Studio e la Cura delle Malattie Infiammatorie Croniche Intestinali, University of Pavia, Pavia; 5 Centre for Immunology and Infectious Disease, Bicms, Barts and The London School of Medicine and Dentistry, London, UK
Background and aim: While interleukin (IL)-17A is pro-inflammatory, IL17E (also known as IL-25) is anti-inflammatory in Crohn’s disease (CD). There are no studies investigating the role of IL-17E in modulating extracellular matrix in the gut, while it is known that IL-17A increases matrix metalloproteinase (MMP) production by intestinal fibroblasts. Here we have studied the role of both IL-17A and IL-17E in CD fibrogenesis. Material and methods: Fibroblasts were isolated from colonic surgical specimens collected from strictured and non-strictured areas of 10 patients with fibrostenotic CD, and normal areas of 10 control subjects. The expression of IL-17R, which is common to both IL-17A and IL-17E, was determined on fibroblast lysates by immunoblotting. Fibroblasts were cultured for 24 hours with recombinant human IL-17A and IL-17E. Then supernatants were used for detection of soluble collagen by Sircoll Collagen Assay and tissue inhibitor of MMPs (TIMP-1) by immunoblotting. Fibroblast migration was assessed using the in vitro wound-healing scratch assay. Results: IL-17R was expressed on CD strictured, CD non-strictured and control fibroblasts, with no significant differences between the three cell populations. IL-17A but not IL-17E significantly increased collagen production and TIMP-1 expression, mainly in the supernatants of CD strictured fibroblasts. Migration of CD strictured, CD non-strictured and control fibroblasts was significantly inhibited by IL-17A but not IL-17E, with no significant differences between the three cell populations. Conclusions: Our results suggest that IL-17A but not IL-17E is profibrotic in CD. Further studies are needed to clarify the molecular mechanisms underlying this profibrotic action. # G. Inflammatory bowel diseases - 1. Basic science
P.64 INTERLEUKIN-25 PRODUCTION IS DIFFERENTLY REGULATED BY TNF-α AND TGF-β1 IN THE HUMAN GUT D. Fina ∗ ,1 , E. Franzè 1 , L. Rovedatti 2 , G. Corazza 2 , P. Sileri 3 , T. MacDonald 4 , F. Pallone 1 , A. Di Sabatino 2 , G. Monteleone 1 1 Dipartimento
di Medicina Interna, Università “Tor Vergata”, Roma; Medica I, Fondazione IRCCS Policlinico, San Matteo, Università di Pavia, Pavia; 3 Dipartimento di Chirurgia, Università “Tor Vergata”, Roma; 4 Institute of Cell and Molecular Science, Barts and The London School of Medicine and Dentistry, London 2 Clinica
Background and aim: IL-25 has been recently shown to suppress the production of inflammatory cytokines by gut antigen presenting cells, and to inhibit Th1 cell-driven inflammation in experimental colitis. Interestingly, a diminished production of IL-25 occurs in the inflamed gut of IBD patients. These findings raise the possibility that IL-25 down-regulation can contribute to amplify and/or maintain the ongoing tissue-damaging immune response in IBD. Factors/mechanisms underlying the defective production of IL-25 in IBD remain however unknown. We investigated whether the decreased production of IL-25 is a hallmark of IBD, and which factors/mechanisms control IL-25 production. Material and methods: IL-25 was examined in mucosal samples taken from IBD, celiac disease, and normal controls by ELISA. IL-25 was also mea-
P.62 INFLIXIMAB MODULATES MUCOSAL CYTOKINE PROFILE IN PATIENTS WITH INFLAMMATORY BOWEL DISEASES F. Caprioli ∗ ,1 , F. Bosè 2 , L. Raeli 2 , C. Viganò 3 , G. Basilisco 1 , D. Conte 3 , S. Abrignani 2 , E. Reali 2 1 Gastroenterology
Unit 2. Fondazione IRCCS Ospedale Maggiore Policlinico, Mangiagalli e Regina Elena, Milano; 2 INGM-National Institute of Molecular Genetics, Milano; 3 Department of Medical Sciences, Università degli Studi di Milano, Milano Background and aim: Monoclonal antibodies against tumor necrosis factoralpha (TNFα) are highly effective in the treatment of patients with inflammatory bowel diseases (IBD), even if their mechanism of action has yet to be fully elucidated. In patients with psoriasis, TNFα neutralization reduces dermal expression of proinflammatory cytokines produced by Th1 and Th17 lymphocytes, T cell lineages critically involved also in the pathogenesis of IBD. In this study, we aimed to evaluate the effect of TNFα neutralization by infliximab in modulating mucosal cytokine profile in patients with IBD. Material and methods: Twelve patients with active left-sided colonic IBD (6 patients with Crohn’s disease (CD) and 6 with ulcerative colitis, 7M, median age 34 yrs, range 25-50), and naïve to anti-TNF therapy were enrolled in the study. All patients underwent induction therapy with infliximab (5 mg/kg administered at week 0, 2 and 6). At enrolment and at week 6 a sigmoidoscopy was performed, and samples were obtained from the sigmoid colon. RNA was extracted from mucosal samples and quantitative expression of 96 inflammation-related genes was evaluated by qPCR, by means of TaqMan Low Density Array Human Immune Panel (Applied Biosystems). Mucosal gene expression was evaluated before and after therapy, and compared with results obtained from 16 controls with normal-appearing mucosa (6M, median age 38 yrs, range 20-44). Results: Among the panel of 96 inflammation-related genes, 36 were found to be significantly upregulated in the mucosa of patients with active IBD in comparison to controls. As compared to baseline, 6 weeks therapy with infliximab induced a significant downregulation of the expression of interleukin (IL)17A, IL-1β, IL-1α, IL-6, IL-8 and NOS2A (all P<0.05). In the CD patients subgroup, TNFα neutralization led to a reduced expression of interferongamma (IFNγ) and lymphotoxin-α, and to an upregulation of the expression of IL-4. With the exception of IL-8 and NOS2A, no significant differences were observed between the post-therapy levels of the above-mentioned genes and those measured in controls. Conclusions: Our data demonstrate that TNFα neutralization rapidly normalizes mucosal expression of several inflammation-related genes, including the signature Th1 and Th17-cytokines IFNγ and IL-17A. These results provide further insights on the mechanisms of anti-TNF agents in controlling mucosal inflammation. # G. Inflammatory bowel diseases - 1. Basic science
S125
P.63 INTERLEUKIN (IL)-17A BUT NOT IL-17E IS PROFIBROTIC IN CROHN’S DISEASE L. Rovedatti ∗ ,1 , A. Di Sabatino 1 , F. Vidali 1 , D. Fina 2 , P. Biancheri 1 , C. Ubezio 1 , A. Pasini 1 , G. Sampietro 3 , C. Alvisi 4 , M. Perego 4 , D. Foschi 3 , T. MacDonald 5 , G. Monteleone 2 , G. Corazza 1 1 First Department of Medicine, Fondazione IRCCS Policlinico S. Matteo, Centro per lo Studio e la Cura delle Malattie Infiammatorie Croniche Intestinali, University of Pavia, Pavia; 2 Department of Internal Medicine, University Tor Vergata, Roma; 3 Surgery Division, Department of Clinical Sciences, Luigi Sacco University Hospital, Milano; 4 Endoscopy Unit, Fondazione IRCCS Policlinico S. Matteo, Centro per lo Studio e la Cura delle Malattie Infiammatorie Croniche Intestinali, University of Pavia, Pavia; 5 Centre for Immunology and Infectious Disease, Bicms, Barts and The London School of Medicine and Dentistry, London, UK
Background and aim: While interleukin (IL)-17A is pro-inflammatory, IL17E (also known as IL-25) is anti-inflammatory in Crohn’s disease (CD). There are no studies investigating the role of IL-17E in modulating extracellular matrix in the gut, while it is known that IL-17A increases matrix metalloproteinase (MMP) production by intestinal fibroblasts. Here we have studied the role of both IL-17A and IL-17E in CD fibrogenesis. Material and methods: Fibroblasts were isolated from colonic surgical specimens collected from strictured and non-strictured areas of 10 patients with fibrostenotic CD, and normal areas of 10 control subjects. The expression of IL-17R, which is common to both IL-17A and IL-17E, was determined on fibroblast lysates by immunoblotting. Fibroblasts were cultured for 24 hours with recombinant human IL-17A and IL-17E. Then supernatants were used for detection of soluble collagen by Sircoll Collagen Assay and tissue inhibitor of MMPs (TIMP-1) by immunoblotting. Fibroblast migration was assessed using the in vitro wound-healing scratch assay. Results: IL-17R was expressed on CD strictured, CD non-strictured and control fibroblasts, with no significant differences between the three cell populations. IL-17A but not IL-17E significantly increased collagen production and TIMP-1 expression, mainly in the supernatants of CD strictured fibroblasts. Migration of CD strictured, CD non-strictured and control fibroblasts was significantly inhibited by IL-17A but not IL-17E, with no significant differences between the three cell populations. Conclusions: Our results suggest that IL-17A but not IL-17E is profibrotic in CD. Further studies are needed to clarify the molecular mechanisms underlying this profibrotic action. # G. Inflammatory bowel diseases - 1. Basic science
P.64 INTERLEUKIN-25 PRODUCTION IS DIFFERENTLY REGULATED BY TNF-α AND TGF-β1 IN THE HUMAN GUT D. Fina ∗ ,1 , E. Franzè 1 , L. Rovedatti 2 , G. Corazza 2 , P. Sileri 3 , T. MacDonald 4 , F. Pallone 1 , A. Di Sabatino 2 , G. Monteleone 1 1 Dipartimento
di Medicina Interna, Università “Tor Vergata”, Roma; Medica I, Fondazione IRCCS Policlinico, San Matteo, Università di Pavia, Pavia; 3 Dipartimento di Chirurgia, Università “Tor Vergata”, Roma; 4 Institute of Cell and Molecular Science, Barts and The London School of Medicine and Dentistry, London 2 Clinica
Background and aim: IL-25 has been recently shown to suppress the production of inflammatory cytokines by gut antigen presenting cells, and to inhibit Th1 cell-driven inflammation in experimental colitis. Interestingly, a diminished production of IL-25 occurs in the inflamed gut of IBD patients. These findings raise the possibility that IL-25 down-regulation can contribute to amplify and/or maintain the ongoing tissue-damaging immune response in IBD. Factors/mechanisms underlying the defective production of IL-25 in IBD remain however unknown. We investigated whether the decreased production of IL-25 is a hallmark of IBD, and which factors/mechanisms control IL-25 production. Material and methods: IL-25 was examined in mucosal samples taken from IBD, celiac disease, and normal controls by ELISA. IL-25 was also mea-