89 Preoperative Wireless Capsule Endoscopy Does Not Predict Long-Term Outcome After Ileal Pouch-Anal Anastomosis Zuri A. Murrell, Eric A. Vasiliauskas, Marla C. Dubinsky, Andrew Ippoliti, Gil Y. Melmed, Dermot P. McGovern, Stephan R. Targan, Phillip Fleshner PURPOSE: Pouchitis is a common complication following ileal pouch anal anastomosis (IPAA) for ulcerative colitis (UC) and indeterminate colitis (IC). The extent of preoperative small bowel mucosal inflammation may be an important predictor of outcome after IPAA. Wireless capsule endoscopy (WCE) appears to be more sensitive for the assessment of mucosal lesions than any radiological test. The purpose of this study was to investigate the value of preoperative WCE in predicting long-term outcome of IPAA in patients who carry a diagnosis of UC or IC. METHODS: 47 patients (32 UC patients and 15 IC patients) undergoing complete WCE before IPAA with at least a 3 month follow up over a 7-year period ending July 2007 were identified. IC features included atypical distribution of disease, presence or history of perianal disease, small bowel inflammation more than 3 cm proximal to the ileocecal valve, or by identification of transmural inflammation or granulomas in the resected colon. Findings on WCE were classified as positive (erosions, ulcers, erythema), negative, or incomplete. Long-term outcome was assessed prospectively and included no pouchitis (NP), acute pouchitis (antibiotic responsive), chronic pouchitis (antibiotic depend- ent/resistant), or the development of de novo Crohn's disease (CD). Patients with acute pouchitis (AP), chronic pouchitis (CP) or Crohn's disease (CD) were considered to have pouch inflammation (PI). RESULTS: The study group had a median age of 36 years (range, 9 to 73 years), and included 23 males and 24 females. Median follow up time after ileostomy closure was 9 months (range, 3 to 60 months). WCE was positive (WCE+) in 14 patients (30%) and negative (WCE-) in 33 patients (70%). Within WCE+ patients, 1 (7%) developed AP, 1 (7%) developed CP and 3 (21%) developed CD. Within WCE- patients, 6 (18%) developed AP, 2 (6%) developed CP and 2 (6%) developed CD. There was no significant difference in AP, CP, CD or PI between WCE+ patients and WCE- patients. Seven of the 32 UC patients (22%) were WCE+ compared to seven of the 15 IC patients (47%) (p= 0.10). There was no significant difference in PI between the WCE+ and WCE- groups in both the UC (29% vs.36%, p=ns) and IC (43% vs. 13%, p=0.28) patient subgroups. CONCLUSION: This study suggests that there is no significant advantage of using preoperat- ive WCE to predict pouch inflammation after IPAA. 90 Family History of Crohn's Disease Increases the Risk for the Development of Crohn's Disease of Ileal Pouch-Anal Anastomosis Bo Shen, Feza H. Remzi, Bret Lashner, Elaine Queener, Victor W. Fazio Background: Crohn's disease (CD) of the pouch can occur in patients with restorative proctocolectomy and ileal pouch-anal anastomosis originally performed for a pre-operative diagnosis of ulcerative colitis (UC). CD of the pouch was often observed in patients with a family history of CD. The aim of the study was to determine whether the family history of CD increased the risk for the development of CD of the pouch in patients who underwent restorative proctocolectomy. Methods: A total of 558 eligible patients seen in our subspecialty Pouchitis Clinic were prospectively enrolled from 2002- 2007, including 116 patients with CD of the pouch and 442 patients with a normal pouch or other pouch disorders (including pouchitis, cuffits, and irritable pouch syndrome). Demographic and clinical variables were included in the study. Multivariable logistic regression analyses were performed. Results: The adjusted multivariable logistic analyses revealed that the risk for the development of CD of the pouch was increased in patients with a family history of CD with odds ratio (OR) of 3.61 (95%CI 1.72-7.57), or with a first-degree relative with CD (OR=4.48, 95%CI 1.57- 12.8), or with a greater number of family members with CD (OR=2.16 per family member, 95%CI 1.27-3.66), adjusting for age, gender, smoking status, duration of IBD, and duration of having a pouch. In addition, patients with a younger age and longer duration of having a pouch had a higher risk for the development of CD of the pouch. A diagnosis of CD of the pouch was associated with a poor outcome with a 7-fold increased risk for pouch failure (OR=7.08 and 95% CI, 3.21 -15.6). Conclusions: The presence of a family history of CD is associated with an increased risk for the development of CD of the pouch, which in turn has a high risk for pouch failure. The presence of a family history of CD in patients with underlying UC may have an impact on decision on whether restorative proctocolectomy should be performed. Risk Factors for Crohn's Disease of the Pouch-Multivariable Analysis A-15 AGA Abstracts 91 Generation and Characterization of a Novel Nf-κB Reporter System to Study Bacteria-Host Interactions in the Zebrafish Intestine Marcus Muehlbauer, Michelle Kanther, Christian Jobin, John F. Rawls Introduction: The NF-κB family of transcription factors integrates microbial and physiolo- gical stimuli in the gut, and improper activation of this pathway is associated with inflammat- ory disorders including IBD. Our transgenic mouse expressing EGFP under transcriptional control of NF-κB has provided useful information on bacteria-host interactions in the gut, however this system is not suitable for chemical and genetic screens. We recently established a gnotobiotic zebrafish model to investigate bacteria-host relationships in the gut. Aim: Generate a zebrafish NF-κB reporter system to study bacteria-host interactions. Methods: We performed microarray-based functional genomic comparisons in zebrafish larvae raised germ-free versus those colonized with a zebrafish gut microbiota. We generated a reporter plasmid that permits expression of EGFP under control of consensus NF-κB binding sites (pNFKB:EGFP). Zebrafish PAC2 embryonic fibroblasts were transfected with pNFKB:EGFP, and stimulated with LPS or Pseudomonas aeruginosa lysate. Expression of the active NF-κB subunit RelA and the inhibitory protein IκBα was evaluated by Western blot analysis, and transcription of NF-κB target genes was monitored by quantitative RT-PCR. We used the Tol2 transposon system to create a transgenic zebrafish carrying the pNFKB:EGFP reporter (Tg(NFKB:EGFP)), and then monitored EGFP expression In Vivo as a function of microbial status. Results: We identified zebrafish genes that are transcriptionally regulated by the gut microbiota, many of which have mammalian homologs that are involved in innate immunity and regulated by NF-κB. Although the zebrafish genome encodes components of the NF- κB pathway, their functions have not been established. Treatment of PAC2 cells with LPS stimulated RelA nuclear translocation and expression of NF-κB targets such as IκBα. PAC2 cells transfected with pNFKB:EGFP showed enhanced EGFP expression following stimulation with LPS or bacterial lysate. Exposure of zebrafish larvae to the NF-κB inhibitor BAY11072 caused a reduction in IκBα mRNA levels. Having shown that the NF-κB system is functional in zebrafish, we next generated a Tg(NFKB:EGFP) zebrafish line. Colonization of germ-free Tg(NFKB:EGFP) zebrafish with a normal microbiota or P. aeruginosa resulted in elevated EGFP expression in the liver and a distinct population of intestinal cells. Conclusions: Our results indicate that the zebrafish NF-κB pathway is active and capable of responding to microbial stimuli. The optical transparency of the zebrafish provides new opportunities for investigating mechanisms underlying temporal and spatial control of the NF-κB response to gut bacteria. 92 T Cell Signaling Via the Toll-Receptor Associated Activator of Interferon (TRIF) Pathway Protects from the Development of Acute and Chronic Murine Colitis Keith J. Breglio, Masayuki Fukata, Daisy E. Conduah, Yasmin Hernandez, Anli Chen, Tyralee Goo, David Hsu, Ruliang Xu, Maria T. Abreu Introduction: Abnormal T cell responses to commensal bacteria are involved in the pathogen- esis of inflammatory bowel disease (IBD). Recognition of bacteria is dependent on toll-like receptors (TLRs) in the intestinal mucosa. Most TLRs use myeloid differentiation-88 (MyD88) as an adapter molecule for signaling whereas TLR3 and TLR4 use TRIF as an alternative activation pathway. We tested the hypothesis that signaling via TRIF protects against the development of colitis in both acute and chronic models. Methods: Acute colitis was induced by 2.5% dextran sodium sulfate (DSS) for 7 days. Chronic colitis was established by transfer of CD4+CD62L+ naïve T cells from TRIF mutant (LPS2) mice and WT spleens to RAG1-/- mice and followed for 9 weeks. TLR expression was examined by flow cytometry. Severity of colitis was assessed via colonoscopy, change in body weight, and histological score. T cell proliferation and cytokine expression were assessed via 3H-Thymidine and ELISA assays, respectively. Results: In the acute model, TRIF-/- mice showed significantly more weight loss at the end of 7 days of DSS treatment as compared to WT mice (p<0.05). Compared to the transfer of WT naïve T cells, transfer of TRIF-/- T cells resulted in significantly more weight loss in the RAG1-/- recipient mice within the first 3 weeks after transfer. TRIF- transferred mice showed increased inflammatory cell infiltration and crypt architectural distortion on histology as well as more severe inflammation endoscopically. Naïve TRIF mutant T cells exhibit increased proliferation compared to naïve WT T cells. Lamina propria T cells from RAG1-/- mice given TRIF mutant T cells showed increased expression of IL- 17 as compared to WT T cells. Conclusion: TLR signaling via TRIF on T cells plays a role in the protection from colitis in both acute and chronic models of murine IBD. The absence of the TRIF signaling complex results in preferential Th17 T cell differentiation, which may be the mechanism for increased inflammation. These results suggest a role for TLR signaling by T cells in the regulation of mucosal T cell responses and their contribution to IBD. AGA Abstracts