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Pathogenesis of IBDW1-W7 BriishSociet ofGastoelogy Si Pathogenesis ofIBD W1-W7 Wi W3 CIRCULATING LYMPHOCYTES BUT NOT MONOCYTES HAVE DEPRESSED HLA-DR ANTIGEN EXPRESSION ON CIRCULATING Gut: first published as 10.1136/gut.33.2_Suppl.S1 on 1 January 1992. Downloaded from RAISED IL-2 RECEPTOR EXPRESSION IN CROHN'S MONOCYTES IN ACTIVE INFLAMMATORY BOWEL DISEASE (IBD). DISEASE K.R. Gardiner. A.D. Crockard. M.I.Halliday. B.J. Rowlands. Depts M J Weldon. T Poulton. J D Maxwell (Dept.of of Surgery and Immunology, Queen's University of Belfast, N.I. Biochemical Medicine and Immunology. St George's Expression of the human leucocyte antigen DR (HLA-DR) on Hospital Medical School, London.) peripheral blood monocytes correlates with the development of infection and predicts outcome after trauma and surgery. In vitro activated lymphocytes and macrophages We investigated HLA-DR antigen expression on circulating express interleukin 2 receptor (IL-2r) on their monocytes in IBD patients with rcspect to disease activity and cell membranes. IL2r has been detected on bowel outcome. Patients with a clinical relapse of Crohn's disease (CD) or mucosal lymphocytes and macrophages in inflammatory ulcerative colitis (UC) were considered cligiblc for study if bowel disease and the soluble receptor (sIL-2r) they had not undergone surgery or received iniimuno- is raised in serum and correlates with disease suppressants within the previous 3 months. The % positivity activity. This receptor may therefore be important and fluorescent intensity (FI) of expression or HLA-DR antigens on monocytes were determinced M wliole blood in immunopathogenesis. Since there is evidence samples using dual colour immuno-iluorcsccnce lahclling and that circulating monocytes are activated in flow cytometry. Serum alpha 1 acid glycoprotein Crohn's disease, we sought the presence of IL-2r concentration (aGP) was measured as an index of disease on circulating monocytes in Crohn's disease. activity. Asymptomatic voluntecrs scrved as controls. were compared according to diagnosis or CD) wid Using two colour Fluorescein activated cell Patients (UC serum aGP concentration (0.33-0.88 g/l = inactive; >0.88 g/l = sorter analysis of lysed whole blood, IL-2r active). Data expressed as mean ± SEM; significancc vs coitirol expression was measured on both lymphocytes and *=p<O.05; active vs inactive t=p<0.05; Student's t test. monocytes in 15 patients with Crohn's disease and UC CD Active Inactive Control in 6 normal controls. Purity of monocytes examined n= 13 n=5 n=9 n=9 n= 15 % 68.5±5.7 67.2±6.1 57.8±4.9*t 78.5±5.5 73 4±4.0 was over 90% using the monocyte marker CD14. F I 44.9±2.9 45.2±7.1 39.8±2.5t 50.2±4.3* 42.3±1.4 Disease activity was measured using the Harvey Seven patients required surgical intervention within 6 weeks Bradshaw index (HBI). of initial assessment (DR % 52.8±4.5) and 11 reccived only Monocytes showed no expression of IL-2r medical therapy (DR % 77.9±4.5; p=0.002). suggesting that this occurs only on arrival at the This study demonstrates that (1) the % of monocytes displaying HLA-DR positivity and the level of HLA-DR inflamed bowel mucosa. However, expression of expression were significantly decreased in patients with active lymphocyte IL-2r was significantly raised in both disease and (2) in patients requiring surgical resection of active and inactive Crohn's disease (p<0.01) diseased bowel, the % of monocytes displaying HLA-DR compared to normal controls and correlated with positivity was significantly reduced compared with patients measured by HBI (r=0.7, p<O.01). receiving mcdical therapy alone. The measurement of HLA- disease activity DR exprcssion on circulating monocytes may prove useful in This study provides further evidence for systemic predicting outcome of IBD relapses. immune activation in Crohn's disease. http://gut.bmj.com/ W2 W4 P:.N A PCHACGtP E'T IVP. I ON INStCSU 0' t FI ' E t H PrJ',0 S 1I -5NC!J L T'P C H l rN- 1 L REACTIVE OXYGEN METABOLITES DAMAGE DNA IN SMALL( IEsTF TINE. INFLAMMATORY BOWEL DISEASE. NJ Simmonds. P tt.tt .- H Mur h J. Wri)' w t -A' on September 24, 2021 by guest. Protected copyright. S Bashir. G Harris. JCW Lee. PG Winyard. DR Blake. DS Rampton. macDonal.-i_ Inflammation Group, Gastrointestinal Science Research Unit, 26 Ce p).¢ ement !f Paedia tric C-:it u.ifl3c .f. t Ashfield Street, London, El 2AJ. Eiarthcionmew'-. Hcspitar. 'Londort. U, DNA of target cells can be damaged by reactive oxygen metabolites. 8- t m ni u n1oP a thCo{) ng lcc. a.l Os 0i ,' t oxo-7-hydrodeoxyguanosine (8-oxodG) is an important product of iontectine o c.n- in twoe rtain t\,pes th e P;apt; ye oxidative DNA damage which is formed by the reaction of the hydroxyl le.sions s-eer i/] cordi tions uch ¢s coe.lia- radical at the CS position of deoxyguanosine. It can result in base 'iise'-. cha ratcr ised by villous atrophy iri changes and point mutations in replicating DNA and so could play a role Cyri hvpe-rr jsia. .rrd destt-LvC 1eoit.ve in carcinogenesis. 8-oxodG can be measured by high performance -( vons' *iseaqe, where mucosal ulceration liquid chromatography with electrochemical detection. We have used a promiuent. feature. In cultured human fet.al this technique to look for evidence of oxidative damage in the peripheral sm.ll intestira1 explanIts we have pr .vioucIy blood and in full-thickness ileocolonic surgical resection specimens of do:sc:r it'e murosal adapt ive changes by .ic ti vat i r patients with inflammatory bowel disease (IBD). ..amina propria CD4+ T cells with pokeweed a. t.ogen (PWM) in the presence of hydrocort ea.,ne Results These are given as the mean and expressed as moles of 8- in-, the mel-iun. In this study, on) i ttinlg oxodG / 106 moles of deoxyguanosine. Significance was assessed co rticot.t-sroids fratn the nmedium we sl-.ow that using analysis of variance. simi lar T cell activation with PWM le3ds to i-apid notos;o at destr uct ion wi th loss c f .i I 1ij . Blood Tissue short hypoplastic crypts and epithelial damage. Lymphocytes (n) Polymorphs (n) (n) Steroid administration is associated wilT.h Controls 62 (31) 116 (28) 143 (6) decreased levels of IL-2 in the cult!;Se Ulcerative colitis 84* (16) 211* (16) 164 (14) supernatant.. In contrast FK 50o and c:yc:lousporine Crohn's disease 60 (7) 154 (11) 255* (10) A completely inhibit T cell activation and the in vitro enteropathy. As demonstrated by * p<0.05 vs. controls (Fisher's PLSD) sequential immunostaining, in the absence of steroids T cell activation leads to the Levels of 8-oxodG were significantly increased in peripheral blood in activation of lamina propria macrophages. ulcerative colitis and in gut in Crohn's disease. Althouigh superoxide anion is produced in increased quantities by these activated Conclusions 1. Increased amounts of 8-oxodG in blood and in tissue macrophages ( and inhibited by superoxide provide evidence of oxidative stress in IBD. 2. Since 8-oxodG is a dismutase ) neither superoxide dismutase nor promutagenic DNA lesion, increased production of this in IBD may play catalase prevent tissue destruction. N-methyl-1- a role in the development of cancer in these diseases. arginine is also ineffective, making it unlikely that nitric oxide plays a role in tissue destruction. Release of proteolytic enzymes is now the most likely mechanism for mucosal destruction in this model. S2 BritishSociety ofGastroenterology W5 W7 REGULATION OF COLONIC EPITHELIAL DERIVED IL-8 CELL* Gut: first published as 10.1136/gut.33.2_Suppl.S1 on 1 January 1992. Downloaded from PRODUCTION OF COLITIS-SPECIFIC MURINE MONOCLONAL SECRETION. C.-C. Schuerer-Maly, F.-E. Maly*. M.F. Kagnoff ANTIBODIES USING A NOVEL TOLERISING TECHNIQUE (introduced by M. E. Parsons). Dept. of Medicine, Laboratory of Mucosal Immunology, University of SBloom 1 Qalifornia, San Diego, School of Medicine, La Jolla, CA and Institute of Molecular Medicine, John Radcliffe Hospital1, Oxford and The Research Institute, Scripp's Clinic and Research British Biotechnology Limited2, Oxford, UK. J . Cytokines produced by intestinal epithelial cells may Murine monoclonal antibodies that recognise antigens specific to be an important signalling. system to the adjacent and Inflammatory bowel disease (lBD) and not found in normal colon have been underlying mucosal immune system. In the present study generated by tolerising mice to normal colonic mucosal cells and then we asked if intestinal epithelial cells produce the potent immunising with cells from inflamed mucosa. neutrophil chemoattractant IL-8. Three colonic Mice were tolerised by 3 cycles of injection with normal colonic cells adenocarcinoma derived cell lines: T84, HT29 and SW620 followed by IP cyclophosphamide to minimise clonal expansion of B cells were stimulated with either phorbol ester (PMA) or the reacting to normal colonic antigens. Disease specific monoclonal antibodies interleukin-1 beta (IL-l)sitr facftor alpha (TNF-a)c were generated by subsequent immunisation with mucosa from Ulcerative beta (TGF-l), interferon-1 (IFNy) and epidermal growth Colitis (2 mice), Crohns disease (2 mice) or a pure population of epithelial factorG.Confluent monolayers were incubated fo 16r hours cells from a case of severe refractory distal colitis where the tolerising in the presence of the agonists (all 0.1-300ng/ml, except cells were from the same patients non-involved right colon. Hybridomas IFNY: 1-100OU/ml). IL-8 secretion into the supernatants were produced using standard techniques and antibodies screened against was determined by ELISA and a bioassay, mRNA frozen sections of colonoscopic biopsies from 15 patients with IBD (11 UC, production was analyzed by Northern blot hybridization.
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