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THE BURDEN OF A CONSERVATIVE APPROACH TO INFLAMMATORY BOWEL DISEASE AT A REFERRAL CENTER
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Abstracts / Digestive and Liver Disease 41S (2009), S1–S167
immunogenic. Their role in experimental colitis has not been explored yet. Aim of this study was to assess A-MSC potential therapeutic role in Dextran Sodium Sulfate (DSS) model of experimental colitis in mice. Material and methods: A-MSCs were isolated from the adipose tissue of C57BL/6 mice. Mesenchimal stem cell properties were evaluated by flow cytometry, assessing CD90, SCA1 and CD49 expression, and by adipocyte and osteocyte potential differentiation, using specific differentiation media. C57BL/6 mice received 2.5% DSS in tap water for 9 days. Cyclosporine (Cy, 10 μg/kg/day) was used as in order to avoid possible rejection. A-MSCs (106 cells) were injected intraperitoneally at day 3. Mice were divided into 4 groups: (A) control mice receiving DSS only; (B) mice receiving DSS+Cy; (C) mice receiving DSS + A-MSC and (D) mice receiving DSS+Cy+ A-MSC. Disease Activity Index (DAI), measured daily, was used to express the severity of colitis together with the change in body weight; survival was expressed as percentage. Results: A-MSC differentiated in osteocytes/adipocytes, and expressed CD90, SCA1 and CD49, showing typical features of mesenchimal stem cells. Once asserted their properties, A-MSCs were used in experimental colitis. Mice treated with A-MSC alone or in combination with Cy showed a significant increase in survival compared to control mice (80 and 100% vs 50%). DAI as well as loss in body weight were reduced in mice receiving A-MSC in combination with Cy compared to controls (p < 0.05 at day 7, 8, 9). Any other significant differences were not observed. Conclusions: A-MSCs were able to attenuate the severity of DSS colitis in mice, as shown by survival, DAI and body weight loss. Further experimental data are required to confirm these results and to assess the mechanisms of action of A-MSC. # L. Inflammatory bowel diseases 1. Basic science
P.134 CHARACTERIZATION OF IL-21 PRODUCING CELLS IN INFLAMMATORY BOWEL DISEASE M. Sarra ∗ ,1 , I. Monteleone 1 , C. Stolfi 1 , L. Biancone 1 , G. Sica 2 , P. Sileri 2 , R. Tersigni 3 , F. Pallone 1 , G. Monteleone 1 1 Università
Tor Vergata - Cattedra di Gastroenterologia, Roma; Tor Vergata - Cattedra di Chirurgia Generale, Roma; 3 Azienda Ospedaliera S. Camillo-Forlanini-Spallanzani, Roma 2 Università
Background and aim: In Inflammatory bowel disease (IBD) the tissue lesions occur in areas which are massively infiltrated with cytokinessecreting mononuclear cells. We have previously demonstrated that in the inflamed gut of patients with Crohn’s Disease (CD) and patients with Ulcerative Colitis (UC) there is enhanced production of IL-21, and that this cytokines contributes to expand multiple inflammatory pathways. Although, IL-21 has been traditionally considered a product of activated CD4+ T cells and activated NKT cells. T Follicular helper (TFh) cells, expressing CXCR5, can also make IL-21. The aim of this study was to characterize the cell sources of IL-21 in IBD and to examine whether IL-21 producing cells co-express other cytokines. Material and methods: LPMC were isolated from mucosal sample of patients with CD, patients with UC and Normal Controls (NC) and used to assess IL-21 producing cells or to purify CD4+ T Lamina Propria Lymphocytes (T-LPL). IL-21-producing cells were also examined in Peripheral Blood Mononuclear Cells (PBMCs). CD4+ T-LPL were stimulated with anti CD3 + CD28 antibodies in presence or absence of IL-12 or IL-4. Cytokines expression was assessed by flow-cytometry. Results: The fraction of IL-21-expressing CD4+ T-LPL was increased in CD as compared to UC and NC. Moreover, IL-21-producing CD4+ T-LPL were more frequent in UC than in NC. By contrast there was no difference in terms of IL-21-expressing cells in PBMC of IBD patients and normal controls. In both CD and UC, more than 50% of IL-21producing cells co-expressed IFN-gamma and/or CXCR5, while less
than 15% co-expressed IL-4 or IL-17A. In line with this, stimulation of CD CD4+ T-LPL with IL-12, the major Th1 inducing factor, enhanced the percentage of IL-21 and IFN-gamma producing cells, as well as the fraction of cells co-expressing both cytokines. By contrast, stimulation of UC CD4+ T-LPL with IL-4 did not modify the fraction of IL-21 producing cells. Conclusions: In IBD IL-21 is mostly produced by CD4+ T-LPL coexpressing IFN-gamma. Data reinforce the concept that IL-21 is made by Th1 cells in human IBD. # L. Inflammatory bowel diseases 1. Basic science
P.135 THE BURDEN OF A CONSERVATIVE APPROACH TO INFLAMMATORY BOWEL DISEASE AT A REFERRAL CENTER G. Actis, P. Pazienza, M. Ayoubi, G. Tappero, F. Rosina ∗ Presidio Sanitario Gradenigo, Torino Background and aim: Cyclosporin, azathioprine, and mesalamine are still acknowledged to form the backbone for treatment of moderatesevere inflammatory bowel disease. The impact of these cost-effective drugs is being strongly emphasized in these times of financial restriction, yet it may be significantly limited by toxicity forcing withdrawal. We addressed this issue taking advantage of the real-world experience gathered at our referral center. Material and methods: We studied 94 patients: 79 with ulcerative colitis (57% had pancolitis) and 15 with Crohn’s disease (gastricileal-colonic in 60%). Relevant data comprised 63 treatments with cyclosporin (2mg/kg intravenously or 5 mg/kg orally); 57 with azathioprine (2 mg/kg); and 44 with mesalamine (3.2-4.8 gr). After the 14-day induction phase, oral cyclosporin was continued for up to 6 months; the median azathioprine duration was 14 months (1-201); mesalamine was meant to be continued until tolerated. Results: CsA: toxicity frequency:16/63 (25%); fatality 1.6%; renal damage 14%; liver toxicity 8%. Three of the 5 cases of liver injury were withdrawn and colectomized. AZA: toxicity rate: 25/57 (43%) with 27 events, including leukopenia (17.5%), liver damage (8.7%) and infection (7%) at the top three ranks. Overall time-to-toxicity: 1-60 months (median 6); drug discontinuation required by 14/25 (56%). 7/18 UC patients showing toxicity had colectomy. 62% of the events were other than leukopenia. 5-ASA: toxicity rates: 6/44 (13.6%) with 7 events. Lung hypersensitivity ranked first: three patients (9%). All the three were withdrawn and one needed colectomy. Conclusions: Conventional induction and maintenance regimes for inflammatory bowel disease are plagued with significant side-effects requiring withdrawal often followed by severe relapses. >50% of the thiopurine toxicity depends on as yet unidentified metabolic polymorphisms that outrange the power of the available pharmacogenomic assays; unexpectedly, allergic lung dysfunction was the top-list undesired event of mesalamine in this series. Efforts are needed to optimize the use of these two drugs that make the cheapest mainstay for the control of inflammatory bowel disease. # L. Inflammatory bowel diseases 1. Basic science
Abstracts / Digestive and Liver Disease 41S (2009), S1–S167
immunogenic. Their role in experimental colitis has not been explored yet. Aim of this study was to assess A-MSC potential therapeutic role in Dextran Sodium Sulfate (DSS) model of experimental colitis in mice. Material and methods: A-MSCs were isolated from the adipose tissue of C57BL/6 mice. Mesenchimal stem cell properties were evaluated by flow cytometry, assessing CD90, SCA1 and CD49 expression, and by adipocyte and osteocyte potential differentiation, using specific differentiation media. C57BL/6 mice received 2.5% DSS in tap water for 9 days. Cyclosporine (Cy, 10 μg/kg/day) was used as in order to avoid possible rejection. A-MSCs (106 cells) were injected intraperitoneally at day 3. Mice were divided into 4 groups: (A) control mice receiving DSS only; (B) mice receiving DSS+Cy; (C) mice receiving DSS + A-MSC and (D) mice receiving DSS+Cy+ A-MSC. Disease Activity Index (DAI), measured daily, was used to express the severity of colitis together with the change in body weight; survival was expressed as percentage. Results: A-MSC differentiated in osteocytes/adipocytes, and expressed CD90, SCA1 and CD49, showing typical features of mesenchimal stem cells. Once asserted their properties, A-MSCs were used in experimental colitis. Mice treated with A-MSC alone or in combination with Cy showed a significant increase in survival compared to control mice (80 and 100% vs 50%). DAI as well as loss in body weight were reduced in mice receiving A-MSC in combination with Cy compared to controls (p < 0.05 at day 7, 8, 9). Any other significant differences were not observed. Conclusions: A-MSCs were able to attenuate the severity of DSS colitis in mice, as shown by survival, DAI and body weight loss. Further experimental data are required to confirm these results and to assess the mechanisms of action of A-MSC. # L. Inflammatory bowel diseases 1. Basic science
P.134 CHARACTERIZATION OF IL-21 PRODUCING CELLS IN INFLAMMATORY BOWEL DISEASE M. Sarra ∗ ,1 , I. Monteleone 1 , C. Stolfi 1 , L. Biancone 1 , G. Sica 2 , P. Sileri 2 , R. Tersigni 3 , F. Pallone 1 , G. Monteleone 1 1 Università
Tor Vergata - Cattedra di Gastroenterologia, Roma; Tor Vergata - Cattedra di Chirurgia Generale, Roma; 3 Azienda Ospedaliera S. Camillo-Forlanini-Spallanzani, Roma 2 Università
Background and aim: In Inflammatory bowel disease (IBD) the tissue lesions occur in areas which are massively infiltrated with cytokinessecreting mononuclear cells. We have previously demonstrated that in the inflamed gut of patients with Crohn’s Disease (CD) and patients with Ulcerative Colitis (UC) there is enhanced production of IL-21, and that this cytokines contributes to expand multiple inflammatory pathways. Although, IL-21 has been traditionally considered a product of activated CD4+ T cells and activated NKT cells. T Follicular helper (TFh) cells, expressing CXCR5, can also make IL-21. The aim of this study was to characterize the cell sources of IL-21 in IBD and to examine whether IL-21 producing cells co-express other cytokines. Material and methods: LPMC were isolated from mucosal sample of patients with CD, patients with UC and Normal Controls (NC) and used to assess IL-21 producing cells or to purify CD4+ T Lamina Propria Lymphocytes (T-LPL). IL-21-producing cells were also examined in Peripheral Blood Mononuclear Cells (PBMCs). CD4+ T-LPL were stimulated with anti CD3 + CD28 antibodies in presence or absence of IL-12 or IL-4. Cytokines expression was assessed by flow-cytometry. Results: The fraction of IL-21-expressing CD4+ T-LPL was increased in CD as compared to UC and NC. Moreover, IL-21-producing CD4+ T-LPL were more frequent in UC than in NC. By contrast there was no difference in terms of IL-21-expressing cells in PBMC of IBD patients and normal controls. In both CD and UC, more than 50% of IL-21producing cells co-expressed IFN-gamma and/or CXCR5, while less
than 15% co-expressed IL-4 or IL-17A. In line with this, stimulation of CD CD4+ T-LPL with IL-12, the major Th1 inducing factor, enhanced the percentage of IL-21 and IFN-gamma producing cells, as well as the fraction of cells co-expressing both cytokines. By contrast, stimulation of UC CD4+ T-LPL with IL-4 did not modify the fraction of IL-21 producing cells. Conclusions: In IBD IL-21 is mostly produced by CD4+ T-LPL coexpressing IFN-gamma. Data reinforce the concept that IL-21 is made by Th1 cells in human IBD. # L. Inflammatory bowel diseases 1. Basic science
P.135 THE BURDEN OF A CONSERVATIVE APPROACH TO INFLAMMATORY BOWEL DISEASE AT A REFERRAL CENTER G. Actis, P. Pazienza, M. Ayoubi, G. Tappero, F. Rosina ∗ Presidio Sanitario Gradenigo, Torino Background and aim: Cyclosporin, azathioprine, and mesalamine are still acknowledged to form the backbone for treatment of moderatesevere inflammatory bowel disease. The impact of these cost-effective drugs is being strongly emphasized in these times of financial restriction, yet it may be significantly limited by toxicity forcing withdrawal. We addressed this issue taking advantage of the real-world experience gathered at our referral center. Material and methods: We studied 94 patients: 79 with ulcerative colitis (57% had pancolitis) and 15 with Crohn’s disease (gastricileal-colonic in 60%). Relevant data comprised 63 treatments with cyclosporin (2mg/kg intravenously or 5 mg/kg orally); 57 with azathioprine (2 mg/kg); and 44 with mesalamine (3.2-4.8 gr). After the 14-day induction phase, oral cyclosporin was continued for up to 6 months; the median azathioprine duration was 14 months (1-201); mesalamine was meant to be continued until tolerated. Results: CsA: toxicity frequency:16/63 (25%); fatality 1.6%; renal damage 14%; liver toxicity 8%. Three of the 5 cases of liver injury were withdrawn and colectomized. AZA: toxicity rate: 25/57 (43%) with 27 events, including leukopenia (17.5%), liver damage (8.7%) and infection (7%) at the top three ranks. Overall time-to-toxicity: 1-60 months (median 6); drug discontinuation required by 14/25 (56%). 7/18 UC patients showing toxicity had colectomy. 62% of the events were other than leukopenia. 5-ASA: toxicity rates: 6/44 (13.6%) with 7 events. Lung hypersensitivity ranked first: three patients (9%). All the three were withdrawn and one needed colectomy. Conclusions: Conventional induction and maintenance regimes for inflammatory bowel disease are plagued with significant side-effects requiring withdrawal often followed by severe relapses. >50% of the thiopurine toxicity depends on as yet unidentified metabolic polymorphisms that outrange the power of the available pharmacogenomic assays; unexpectedly, allergic lung dysfunction was the top-list undesired event of mesalamine in this series. Efforts are needed to optimize the use of these two drugs that make the cheapest mainstay for the control of inflammatory bowel disease. # L. Inflammatory bowel diseases 1. Basic science