Ileoanal pull-through improves health related quality of life (HRQL) in patients with a history of ulcerative colitis (UC)

April 1998

Immunology, Microbiology, and Inflammatory Disorders A1045

ileum were confirmed using a multiport sampling enteric tube. In the randomized controlled trial of UCCll8 (101°/day for 21 days), two oral delivery vehicles (milk, n=20 vs yoghurt, n=20; controls n=20 for each delivery vehicle) were then compared for survival/colonisation with UCC118 and its influence on fecal flora• There was no significant difference between milk and yoghurt, in delivery of the UCCll8 lactobacilli, based on viable fecal counts (103-106 organisms/gin feces). Each system was associated with significant increases (10-100-fold) in total fecal lactobacilli, bifidobacteria and enterococci; fecal coliforms and bacteroides were unaltered. Five subjects (5/40; milk 4, yoghurt 1) were still colonized with the UCCll8 strain 21 days after cessation of feeding. No adverse effects were reported. Conclusions: (1) We have developed a probiotic Lactobacillus strain (UCC118), that is anti-microbial in vitro, bifidogenic in vivo, suitable for versatile delivery systems, acid/bile tolerant and survives gut transit in high numbers; (2) in contrast to earlier reports, long term colonization was achieved with this probiotic strain; and (3) the properties of this strain are advantageously suited to future therapeutic testing in IBD. Support: E.U., Bioresearch Ireland & the Health Research Board, Ireland. • G4275 NEUTRALISATION OF ENDOGENOUS TRANSFORMING GROWTH FACTORI31 (TGFI31) PREVENTS INTESTINAL FIBROSIS. M. Mourelle F. Guarner, I-R. Malagelada, Digestive System Research Unit, Hospital General Vail d'Hebron, Barcelona; J. Casalots, A. Salas, Pathology, Hospital Mutua, Terrassa, Spain. Fibrotic strictures may complicate Crohn's disease, colonic ischemia, radiation enteritis or acute diverticulitis. Transmural inflammation appears to be the primary pathological condition leading to fibrosis. We induced intestinal fibrosis in the rat by intramural injection of a suspension of colonic contents, and investigated the role of TGF[31 in this effect. Colonic contents from 4 normal rats were pooled in saline and filtered. Under anesthesia, 4 groups of rats were laparotomized. Sham rats (n = 5) were injected into the colonic wall with 200 laL of sterile saline, and 3 other groups (n = 8, each) with 200 laL of the filtered colonic suspension. The margins of the injection site were then infiltrated subserosally: in the Anti-TGF group with 50 lag of neutraiising antibody to TGFfll (R&D Systems, Minneapolis, Mn), and in the lgG group, with 50 lag of nonspecific lgG. The Control group received no further treatment. Rats were killed 5 days later, and the treated colonic segments were retrieved for assay of tissue collagen and TGFI31 concentration, and for histological examination. Results: * p<0.05 versus Control Sham Control Anti-TGF IgG

TGFI31 (ng/cm2) 0.4 + 0.15" 7.7 ± 1.42 3.9 ± 0.76* 8.2 ± 1.51

collagen (mg/cm2) 0.1 ± 0.03* 1.8 -+0.39 0.6 _+0.20* 1.5 ± 0.40

Intramural injection of the suspension of colonic contents increased tissue TGF~ 1 and induced collagen deposition in the colonic wall. The neutralising antibody to TGFI31 significantly prevented the increase of both tissue TGFI31 and collagen deposition. Histological examination of the colons showed diffuse serosal thickening and fibrosis in Control group, but only minor focal changes in the Anti-TGF group. Treatment with the nonspecific antibody had no effect. Conclusion: Collagen deposition in the colonic wall may be prevented by neutralising antibodies to TGFI31, suggesting that this cytokine plays a pivotal role in postinflammatory intestinal fibrosis. • G4276 ILEOANAL PULL-THROUGH IMPROVES HEALTH RELATED QUALITY OF LIFE (HRQL) IN PATIENTS WITH A HISTORY OF ULCERATIVE COLITIS (UC). A Muir, B Phillips-Bute, M Eloubeidi, C Sears, RR Bollinger, M Koruda, L Hurd, D Bachwich, D Drossman, D Provenzale. Division of Gastroenterology, Department of Surgery, Duke University Medical Center, Durham VA Medical Center, Hospital of the University of Pennsylvania, University of North Carolina-Chapel Hill, NC. HRQL is an important outcome for patients with UC who undergo colectomy. In an earlier analysis, we validated the SF-36, the most widely used measure of HRQL, in post colectomy patients with a history of UC. Specific Aim: To evaluate the change in HRQL pre- to post-colectomy in patients with UC who undergo colectomy with ileoanal pull-through. Methods: Consecutive patients with chronic UC undergoing colectomy for severe colitis or dysplasia were administered the SF-36 pre-operatively and 1 yr post take down (N=17). Results: The mean age was 38, 53% were males and 94% were white. The mean duration of disease pre-operatively was 9 years. SF-36 results (medians and interquartile ranges (IQR)) are shown in the table, along with median scores for a healthy age matched population. The median difference between pre-operative and one-year post-operative measurements was calculated.

Pre-Op Median (IQR)

1 YR Median (IQR)

Healthy

PF

55 (30-90)

95 (85-100)

20 (5-40)

SF

63 (25-75)

100 (75-100)

37.5(0-63) 0.003

I00 (75-100)

RP

25 (0-100)

100 (75-100)

75 (0-75)

100 (100-100)

RE

33 (33-100)

100 (67-100)

0 (0-67)

0.25

100 (67-100)

MH

60 (48-80)

84 (72-88)

16 (8-32)

0.002

80 (64-88) 65 (50-75)

0.0001

0.002

VT

30 (10-60)

65 (45-75)

25 (5-50)

0.002

BP

62 (31-74)

74 (62-84)

12 (0-31)

0.01

HP

32 (25-52)

67 (57-77)

95 (85-100)

84 (62-100) 77 (67-90)

Patient scores improved significantly on all subscales except for functioning related to emotional health (RE), with measures of physical functioning (PF,RP,VT) demonstrating the most dramatic improvement. Clinically, HRQL was poor pre-operatively but at 1 yr was similar to that of a healthy population. Conclusions: 1) Based on our results, HRQL improves dramatically after colectomy in patients with chronic UC, with the most dramatic improvement in physical function. 2) The SF-36 is responsive to changes in HRQL related to surgery in UC patients and can be used to measure HRQL in future trials. 3) HRQL was poor pre-operatively but at 1 yr was similar to that of a healthy population. 4) Our results provide new data on clinical outcomes post-colectomy that can be used by physicians and patients to estimate the outcome of ileoanal pull-through for those considering surgery. 5) Policy makers can use these results to incorporate HRQL into strategies that will maximize this outcome for patients with UC. Supported in part by a grant from the American College of Gastroenterology. G4277 CROHN'S DISEASE ANTIGEN PRESENTING CELLS INDUCE INCREASED IFNT VS. ULCERATIVE COLITIS. GE Mullin, J Costable, R Cerchia, T Bui, L Stiel, C Werner, J Procaccino, C Chang and M Chang. Department of Medicine, North Shore University Hospital, NYU School of Medicine, Manhasset; NY. BACKGROUND: Both histopathologic and mRNA studies suggest that Crohn's disease (CD) has a Thl-like (high IL-2, IFNT) lymphokine profile. The mechanisms responsible for driving this response is unclear. The aim of our study was to determine if CD mucosal antigen presenting cells APCs induce normal peripheral blood T cells (NTs) to secrete increased IFNT. METHODS: Intestinal surgical specimens were obtained from 13 normal Control, 9 ulcerative colitis (UC) and 11 Crohn's disease (CD) patients. Lamina propria mononuclear cells (LPMCs) and intestinal epithelial ceils (IECs) were prepared by a standard method of enzyme digestion with collagenase and dispase (respectively) and percoll gradient centrifugation. Intestinal APCs (iAPCs) were obtained from the T cell depleted fraction of LPMCs by the use of immunomagnetic beads conjugated with anti-CD2 mAb. IECs and iAPCs were cultured with or without normal NTs in duplicate @ at a concentration of lxl06 cells/ml with either; media alone or 10 ng/ml toxic shock syndrome toxin (TSST) for 48 hrs @ 37°C. Cellular proliferation was determined by pulsing cultures with tritium (3H) @ 10.0 laCi/ml for 16 hrs followed by harvesting then counting. The concentration of IL-2 and IFNT in samples was determined by ELISA. Results are expressed as the % proliferation of NTs and induction of NT IFNT by iAPCs and IECs relative to Control peripheral blood APCs (NAPCs) (mean +/- SEM). RESULTS: Proliferation of NTs induced by iAPCs were not different between Controls (79.8 +/- 16.7 %), inflamed CD (CDa) (85.8 +/- 16.2 %) and inflamed UC (UCa) (79.9 +/- 12.3) patients. IEC-induced proliferation of NTs by Controls (62.1 +/- 12.8 %), CDa (114.5 +/- 44.7 %) and UCa (89.4 +/- 19.6%) specimens were also not different from each other. In contrast, CDa iAPCinduced IFNT production by NTs (84.2 +/- 21.5 %) was higher than Controls (30.0 +/- 10.2%) (p=0.01) and UCa (36.9 +/- 11.4 %) (p=0.03). However, IEC-induced IFNT production by NTs were not different for Control (66.7 +/20.4 %), CDa (50.2 4-/- 11.3%) and UCa (50.9 +/- 23.9%). After finding that CDa iAPCs induced increased IFNT, we tested culture supematants from 4 UC and 4 CD specimens for IL-2. IL-2 appeared to be higher in CDa (139.5 +/- 17.8%) when compared to UCa (71 +/- 28.2%) (p=0.04), however further testing is required to firmly establish this observation. CONCLUSIONS: Superantigen induced stimulation of NTs by CDa iAPCs increases IFNT production above UCa and Control levels and is not due to heightened proliferative responses. Inflammatory bowel disease (IBD) and n0n-inflammatory Control IECs induce NT proliferation and IFNT production to similar degrees. Further testing of IL-2 and the Th2 (IL-4, IL-5, IL-10) cytokines will determine whether lamina propria APC in the inflamed mucosal lesions of CD can drive Thl responses in this system. Supported in part by the CCFA and a grant from the Litwin's.