A1034 AGA ABSTRACTS

G4233 T-CELL CYTOKINES IN INFLAMMATORY BOWEL DISEASE: INTRACELLULAR DETECTION OF CYTOKINES USING FLOW CYTOMETRY. B. Mascher, P. Schlenke, E.F. Stange* and M. Seyfarth; Institute for Immunology and Transfusion Medicine and *Medical Clinic I, University of Ltibeck, Germany.

t~tiology of inflammatory bowel disease is still unknown. There are several hints speaking for an immunological background. We wanted to investigated this Thl / Th2 hypothesis for the respective types of colitis in IBD patients.- We therefore investigated T-cell cytokines in peripheral blood mononuclear cells (PBMC) and lamina propria mononuclaer ceils (LPMC) from 6 Crohn' disease (CD), 6 ulcerative colitis(UC) patients and 10 healthy controls. PBMC were isolated using standard procedures, then stimulated for five hours with PMA and ionomycin. The ceils were fixed in 4% paraformaldehyd, then lysed in saponin-buffer. Anti-cytokine antibodies and antibodies to surface markers were added at the same time. Cells were finally measured in the FACS. Cytokines of interest were interferon (IFN)r, interleukin (IL) 2 and tumor necrosis factor (TNF) a as Thl cytokines and IL4, IL5 and IL 10 as Th2 cytokines. For staining of surface markers we used CD3 (T-cells), CD8 (cytotoxic T-cells, CD8neg helper T-cells), CD45RA and CD45R0 for naive and memory cells, respectively. As to surface markers, ratio of CD8pos/CD8neg T-cells was not different between PBMC and LPMC. PBMC mainly revealed the naive phenotype (CD45RApos) whereas these cells were greatly decreased in LPMC which mainly displayed the memory phenotype (CD45R0pos). CD45RA pos cells were significantly increased in UC mucosa when compared to controls and CD patients. All measured cytokines were produced to greater extends in memory cells than in naive cells. IL2 was the main cytokine produced in PBMC and reduced in LPMC. It was produced primarily in CD8 neg cells. IL2 production was significantly reduced in CD45RApos and CD8 pos cells in UC mucosa. IFNr was the main cytokine produced in mucosal cells and percentages of positive cells were always higher than in PBMC, it was produced to similar amounts in CDSpos and CD8neg cells. IN LPMC, CD patients produced highest amounts of IFNr, controls intermediate amounts and UC showed weakest production. Th2 cytokines were produced by very few cells, we did not find any differences between LPMC and PBMC nor between disease groups. IL4 was produced to some higher extents than IL5 and IL 10. In our investigations we could find interesting differences between mucosal and blood derived lymphocytes. As to comparison of disease groups, CD patients showed highest amounts of IFNr production and reduction in IL4 which supports the Thl hypothesis.

G4234 SALAZOSULFAPYRIDINE INHIBITS ENDOTHELIAL CELL- DEPENDENT INTERACTIONS WITH NEUTROPHILS INDUCED BY INTERLEUKIN-113. Y. Masui, N. Yoshida, K. Kassai, Y. Naito, T. Yoshikawa. First Department of Internal Medicine, Kyoto Prefectural University of Medicine, Kamigyo-ku, Kyoto 602, Japan.

The microvascular and parenchymal cell dysfunction incurred on inflammatory bowel disease (IBD) has been attributed to activated neutrophils that adhere in postcapillary venules and subsequently emigrate into the interstitium. Neutrophilendothelial cell interactions are mainly mediated by various adhesion molecules, especially endothelial adhesion molecules such as intercellular adhesion molecule-1 (ICAM-1) and E-selectin. Salazosulfapyridine (SASP) has been used for the therapy of IBD. The objectives of this study were to determine effects of SASP and its elements, sulfapyridine (SP), 5-aminosalicylic acid (5-ASA) on surface expression of ICAM-1 and E-selectin. [Materials and Methods] Neutrophils and endotherial cells were isolated from venous blood of healthy adults and human umbilical vein (HUVEC), respectively. Expression of ICAM-1 and E-selectin on IL-1 13 -stimulated HUVEC and neutrophil adherence to IL-1 [3 -stimulated HUVEC were assessed by enzyme immunoassays. In some experimental series, the effects of the SASP, SP and 5-ASA were investigated at a concentration of 10-4-10 -6 M. [Results] IL-1 13 increased surface expression of ICAM-1 and E-selectin on HUVEC and neutrophil adherence to endothelial cells. Pretreatment of endotherial cells with SASP and 5-ASA, but not SP, reduced expression of ICAM-1 and E-selectin on HUVEC and neutrophil adherence to endothelial cells induced by IL-1 13. [Conclusions] These results suggest that SASP and 5-ASA may be effective for the therapy of IBD through inhibiting endothelial cell-dependent interactions with neutrophils.

G4235 ENDOSCOPIC THERAPY FOR INTESTINAL BEH(~ET AND SIMPLE ULCER OF THE ILEUM. M. Matsukawa T. Yamasaki, M. Kurihara. Dpts. of Gastroenterology, Showa University, Toyosu Hospital, Tokyo, JAPAN.

We have studied the relationship between the clinical course and the treatment in patients with intestinal Behcet (IB: 8 cases) and simple ulcer (SU: 12 cases) from 1970 to 1996. We have recognized ulcer(s) of the terminal ileum and colon with intestinal bleeding or abdominal pain in those 20 cases. We have classified them by the therapy into 2 groups (A: treated by

GASTROENTEROLOGY Vol. 114, No. 4

surgical operation -16 cases, B:treated by endoscopic absolute ethanol-spraying and an elementery diet after 4 recurrent cases of operation or 4 initial cases). At initial examination, lesion(s) of IB or SU had showed a deep ulceration with inflammatory polypoid finding of the colon and ileum. This deep ulceration was not cured by medical therapy. In 8 cases of A group which there was recurrence of the abdominal symptoms after the first operation, apparent ulcerations with stenosis near to anastomosis were proved by an examination. There was only a short interval between the recurrence of the symptoms and the second operation for the recurrent ulcers. Therefore, postoperative surveillance for detecting ulcers must be performed within a short period after the first operation. For patients (B group), the deep or shallow ulcer have cured or decreased the size of the ulcer by absolute ethanol-spraying (10ml) for the ulcer under colonoscopy and an elementary diet. Endoscopic therapy with absolute ethanol-spraying and an elementary diet are effective for intestinal Behcet disease and simple ulcer of the ileum.

G4236

IMPROVEMENT IN INFLAMMATORY BOWEL DISEASE-RELATED QUALITY OF LIFE, DAYS OFF WORK AND SOMATIZATION ASSOCIATED WITH GROUP PSYCHOTHERAPY. R.G. Maunder. W.L Lancee, M.J. Esplen, G.R. Greenberg, Departments of Psychiatry and Medicine, University of Toronto, Mount Sinai Hospital, Toronto, Ontario, Canada.

Supportive-expressive group psychotherapy emphasizes realistic appraisal of the functional limitations imposed by 1BD, reflective expression of affect, and mutual support. The purpose of this study is to examine the effects of a 20 week intervention for inflammatory bowel disease on quality of life (QOL) and to determine the feasibility of a randomized controlled trial. Subjects are adults with Crohn's disease or ulcerative colitis who seek psychological support. QOL is measured with the IBD Questionnaire (IBDQ) at the start and end of the intervention. Responders are defined as those whose IBDQ score improves by at least 10 points. By this definition the response rate in this trial is 64% (responders n = 9, non-responders n = 5) and the difference between response and non-response in terms of improvement in QOL is clinically significant (AIBDQ: responders = 26.7, S.D. = 18.8, nonresponders = -13.5, S.D. -- 11.4, p < 0.001). Responders also improve over therapy in decreased days off work/previous 8 weeks (Adays-off: responders = -24.3, S.D. = 29.6, non-responders = 28.3, S.D. = 30.6, p = 0.03), and improvement in somatization measured by the Brief Symptom Inventory (At-score: responders = -4.6, S.D. = 4.8, non-responders = 6.3, S.D; = 3.3, p = 0.001). Responders do not differ from non-responders at the start of therapy in IBD symptoms, days off work/previous 8 weeks, somatization or global psychiatric distress. These results support a potential treatment effect on quality of life and a potential cost-benefit advantage of therapy due to reduced absenteeism. Further study of this intervention is warranted. This research was made possible by a grant from the Crohn's and Colitis Foundation of Canada.

G4237

IN VIVO EXPANSION OF IDENTICAL T CELL CLONES IN COLON AND SYNOVIUM OF A HLA-B27+ PATIENT WITH COLITIS AND SUBSEQUENT ARTHRITIS. E. May, E. Mg_rker-Hermann, K.-H. Meyer zum Biischenfelde, R. Duchmann. I. Dept. of Medicine, Joh. Gutenberg- University, Mainz, Germany.

It has been suggested that bacteria-specific intestinal T cells might migrate to the periphery, crossreact with autologous articular antigens and induce autoimmune arthritis, especially in genetically predisposed HLA-B27+ individuals. Methods: To test this hypothesis, we applied a two-step PCR system using TCRBV and TCRBJ specific primers ("CDR3 spectratyping") and CDR3 sequencing and asked, whether T cell clones with identical TCRB- sequences are present in the synovium and colon of such patients. We here report the TCRAB+ T cell repertoire analysis from 5 sites of the colon, peripheral blood, synovial fluid and synovial membrane of a HLA-B27+ patient who developed arthritis following acute colitis. Results: Most TCRBV-CDR3 spectra from the intestinal samples, synovial fluid, synovial membrane and peripheral blood of the patient showed multiple bands with normally distributed intensities, indicating polyclonal T cell populations. As an exception, there was a single and massively expanded band within the VB 18-spectra from the synovial fluid specimen and the synovial membrane. This band was also present in the spectra of all colonic specimen and the peripheral blood. When the expanded bands were excised from the gel and analyzed in more detail by reamplification with primers specific for VB 18 and the alternatively used 13 JB segments, two JB-specific reactions (JB 1.5 and JB2.1) were positive in all samples. CDR3-sequencing revealed that both products originated from monoclonal T cells. The TCRB-CDR3 regions of the VB 18JB 1.5 and the VB 18JB 21 positive clone showed striking homology on the amino acid level. Conclusions: Our finding that identical T cell clones are expanded in colon and inflamed joints of a HLA-B27+ individual with arthritis and preceding colitis supports the concept that intestinal T cells can migrate to the periphery where they may be critical for the development of arthritis. The striking overrepresentation of the T cell clones in the joint might reflect specific homing effects or enhanced local proliferation. The CDR3