W1363 Pathologic Prognostic Factors and Outcomes in Patients with Colon Cancers with Low Lymph Node Yields Kavitha Rao, Sandfurd Markowitz, Josepb Willis Background: Over the last 10 ),ears it Ms been identified that adequate lymph node dissection is of paramount importance m assessing patient prognosis and need for adjuvant therapy in colonic carcinoma (CC), It is still common for clinicians, however, to be faced with the dilemma of assessing these patmms needs tbr adjuvant chemotherapy based on pathology reports with kss than 12 identified lymph nodes This study analyzes knowu pathologic prognostic indicators and outcomes in CC patients with low lymph node yields. Design: A retrospective study was perfonued on 47 non-rectal colon cancer resection specimens from patients withotu metastases, operated on at a single tertiary care institution during a 20 year penod in which less than 12 lymph nodes were identified. For each case, the number of lymph nodes recovered, T stage, tumor differentiation, pentumoral Crohnsdike inflammatory infiltrate, tumor inu'a-epitbelial lymphocytic infihmte, angiolymphatic invasmn, perineural invasion, extramural venous invasion, desmoplastic stromal response and tumor growth pattern were assesse~d. Only patients with at least 3 years of clinical Mlow-up were included fi'om which disease free survival information was Obtained. Results: The study consisted of 27 left CC, 18 right CC and 2 unknown sites. The median number of lymph nodes recovered was 7 (3-11), The median survival time in months was 67 (22-243), 14 cancers were of high T stage 42 cancers were intestinal type and 5 were mutinous/signet ring type. 40 cancers were moderately differentiated and 7 cases were pcmrly differentiated. 26 neoplasms had a broad-based growth pattern and 21 cases had an infiltrative growth pattern 20 demonstrated angiolymphatic invasion and 7 cases had perineural invasion Cmhnsdike infiltrate was seen in 4 cases and tumor intra-epitbelial lympbe',cytes in 2 cases 4 patients died of metastatic CC. No pathologic features independently identified in these patients. None of these patients with poor outcomes received adjuvant chemotherapy. Conclusion: Our study indicates that pathologic prognostic factors are not a surrogate to adequate lymph node recovery in deternlining the need for adjuvant chemotherapy and predicting outcome in patients w4th CC W1364 in Vivo Cross-Reactivity of Wheat and Rye T-Cell Epitopes in Celiac Disease Robert P Anderson, David Van Heel, Arthur Tatham, Martin Bamardo Derek P Jewell Adrian V Hill Wheat, rye,and barley are tuxtc in celiac disease (CD). R}~ and barley toxicity may be due to homologous pepudes in wheat, rye and barley prolamins that activate specific intestinal T-cells, ~veral wheat ghadin peptides are known to be T-cell epitupes and some of these are encoded by geues fur prolamins in rye (secalins). It has not been shown tha rye secalins aetivate T-cells in CD, nor have T cells induced by rye exposure in vivo been characterised, AIM: Define wheat gfiadin peptides recogniscd by T-cells induced by in vivo ue challenge in CD, METHODS: HLa,-DQ2 + CD subjects (6) on gluten tree diet ate 4x50g mufhns daily tde 3 days. Muffins were from pure rye flour hand-milled from hand-sorted rye, Blood was collected on day 0, andA~r 6 & 7 (as described in Anderson RP et al Nature Medicine 6:337). Interferon gamma (IFN't) ELIspot assa?~ were pertormed usmg PBMC incubated with a panel of 652 overlapping 20mers spanning all gene-denved Tricitum aestivum c~'~- (61), ',/- (47) and uJ-gliadin (3) sequences in Genbank, and HPLC purified secalin fractions, RESULTS: There was potent induction of secafin-specific IFNy ELlspot responses after ue challenge in 5/6 subiects Each secalin fraction was antigenic (in order of potency: 75kD>40kD>omega secalins; lO0b~g/ml). Deamidation of secalins by tTG increased responds up to 400% lmmunomagnetic bead depletion and anti-dass Ii antibody blocking of PBMC revealed HLA-DQ restricted CD4 T ceils expressing the gnt homing integrin (c~4117) were responslbk for secalin-specific 1FN~/secretion. In 3 subjects with strong secalin-specific ELSipot responses, wheat gliadin peptide specific responses were assessed Gamma- and omega-gliadin peptide responses were dominant and alpha-gliadin peptide (including A- gliadin 57-73) responses were weak, consistent with secahns being homologous to gamma- and omega-gfiadins but not alpha-gliadin Rye challenge induced T cells specific for multiple wheat gamma-gfiadin peptides (most potent: LQQPQQPFPQPQQQLPQPQQ) and omega- gfiadin peptides (most potenu PLQPQQPFPQQPQQPFPQPQ) all nch in Q, P, F and Y CONCLUSION: Rye challenge in vivo induces T-cells specific for both wheat gliadin and rye secalins. Deamidation enhances secalin bioactivity There are multiple wheat gfiadin peptides recognised by T<ells induced by rye exposure in vivo in CD, In contrast to wheat gluten challenge in vivo, after rye challenge gamma- and omega gfiadin peptide-specific IFNy ELIspot responses are substantially" more potent than those to A@iadin 57-73 W1365 A Major Non-HL~ Locus in Celiac Disease Maps to Chromosome 19 Martine Van Belzen Jos W Meqer, Lodewijk A Sandknijl, Afions E. Bardoel, Chris J, Mulder, Peter L. Pearson, Cisca Wijmenga, Roderick H. Houwen Celiac disease (CD) is a multifactoml disorder, characterized by villous atrophy of the small intestine, fi is strongly associated with HLA-DQ2 and DQ8, but other genes are expected to contnbute to CD as well Several genomewide screens have been performed to identify these gs"nes, but none were able to detect genomewide signikam linkage. We therefore performed a genornewide screen in 67 Dutch families with affected sibpairs 1hat met strict diagnostic criteria. Fifteen additional families were available for follow-up of interesting regions The small-intestinal biopsy samples, on which the original CD &agnoses had been based, showed a Marsh I11 lesion in all patients on re-evaluation by one pathologist As expected, highly significant linkage to the: HLA-regmn was detected. More importantly, significant linkage was also present on chromosome 19, with a mufiipoint maximum lod score (MMLS) of 443 (nominal p = 62 x 10~'), The importance of this region was further supported by single point analysis, showing ten consecutive markers in this region with a lod SCore >1, This locus is distinct from other regions reported on chromosome 19 sho',~lng weak evidence tot litfkage. It accounts for a 2 &fold increased risk to siblings, compared to a risk of 46 of the HLA-mgion Furthermore, we identified suggestive linkage to chromo- some 6@1 (MMLS = 2.79, nominal p = 3 x I04). This locus lies ~70 cM downstream from the HLA-region, and has also been implicated in other autoimmune disorders like type 1 diabetes mellitus, suggesting the presence ot art autoimmunity susceptibility gene in this region. Our results clearly demonstrate the presence of a major non-HLA locus involved in CD in the Dutch population This novel, high-risk CD locus is located on chromosome 19p13.1 and is the first non-HLA locus in CD showmg genomewide signihcant linkage. W1366 SNP Mapping of the HLA Class III Region Confirms Existence of an Extended Haplotype in Coe|iac Disease Jacinta Thornton, Jacqueline Daly, Conleth Feigbery, Nicholas Kennedy, Mohamed Abuzakouk, Colm O'Morain, Dermot Kefieher, Ross McManus Background: The majority of northern European celiac patients carry the DQBI*020I allele, however there is tight linkage disequilibrium (LD) in this region and it is possible that other genes may contribute to disease. The objectives of this study were to investigate associations between single nucleotide polymorphisms (SNPs) in genes in the MHC class Ill region and CD, and to ascertain whether these are independent or are in LD with existing HLA associa- tions. We specifically chose coding SNPs or 5"UTR SNPs in genes of potential importance in terms of immunological or signalling function. Methods: 213 patients were included in the study together w{th 1120 randomly selected unaffected individuals as controls. The polymorphisms (SNPs) analysed included 2 coding SNPs (cSNP) in MICB (nucbl and micb2), one cSNP from CSNK2B (csnk2bl), one 5'UTR SNP from DDAH2 (ddah21) and one cSNP from SKW2L (skiv21) Genotyping was performed using Taqman technology" or RFLP. LD was calculated computationally tor the patient group. Allele frequency comparisons were made between patients and controls, Results: Allele frequencies were signiiicantly different between patients and controls for 4 of the 5 SNPs (minor allele frequency shown here): micbl (0,42 vs 072), micb2 (0.15 vs 0.34), ddah21 (0.63 vs 0.32) and skiv2ll (0.36 vs 0.57); p<0 001 in all 4 No diffi'rance was seen in csnk2bl frequencies However, this association could be explained by die significant LD observed between 3 SNPs and HLA DQBI: micbl and HLA~ DQB1 (p=00002), skiv2ll and HLA DQB1 (p=00001) and between ddah21 and HLA DQB 1 (p = 0.001 ). Hapiotypes were also generated computation- ally from this data One haplolype, containing DQBI*0201, was strongly associated with CD Conclusions: The over-representation of alleles in the MHC class Iii region shosved that these polyrnorphisms are highly prevalent in fhe CD population and are strongly linked to DQBI*0201 These po/ymorphisms may also be linked to other susceptibility genes in this region The SNPs studied are known to have immunological, regulatory or signalling {unction and harther analysis of these polymorphisms within tile class lIl region could provide important clues in our understanding of CD pathogenesis W1367 lnterleukin-15 and Intraepithelial Lymphocytes in Celiac Disease Amonio Di Sabatino, Rachele Ciccocioppo, Benedetta Cinque, Alessio Agnifili, Maria Gmzia Cifone, Gmo Roberto Comzza Background & Aims: There is good evidence that intefleukin (IL)-15 has a role in the generation of mucosal damage in celiac disease (CD), and since our previous investigations showed IL-15 overexpression in duodenal mucosa of untreated CD, we investigated ll.-15 production by celiac nitraepitbefial lymphocytes (IELs) to elucidate the role of this pro- inflammatury cytokine in the pathogenesis of CD. Moreover, IL-15 has been reported to manifest an antiapopmtic action by' inhibiting lL-2-mediated-activation induced T-cell apoptosis, and since we previously" demonstrated a defective IEL apoptosts in active: CD, we investigated a putative role of ILd5 in protecting IELs from apoptosis. Patients & Methods: Endoscopic biopsy specimens were obtained from the second part of the duodenum fi:om 10 untl~ated CD patients (mean age 39.2 years, range 19-69), 10 CD patients (mean age 41.5 years, range 20-70) on a gluten-flee diet for at least 12 months, and 10 consenting subjects undergoing upper gastrointestinal eudoseopy for functional dyspepsia (mean age 43.2 years, range 24-70) IEl.s were isolated by mechanical and chemical disruption of the epithelial layer and discontinuous Percoll gradiem centrifugation, and cultured in the absence or presence of IFN-gamma (1000 U/ml). After 24 h of culture, supernatants were aspirated and stored for IL-15 ELISA assay with the Quantikine Human IG15 lmmunoassay (R&D System). To verify a role of IL-15 in protecting IELs from apoptosis, IEks isolated from normal duodenal mucosa were cultured in the absence or presence of recombinant human 1L-15 at different concemrations (1, 5 and 10 ng/ml). Results: IELs isolated t}om untreated celiac mucosa released ILq5 only when stimulated with IFN-gamma The lL-15 levels in the supematants of celiac IFN-gamma-stimulated IELs were significantly higher (mean 28.1 pg/ml, p<O 01) compared with those in the supernatants of treated celiac (mean 6.3 pg/ ml) and control IFN-gamma-stimulated IELs (mean 2.1 pg/ml). IL-15 protects IELs from undergoing apoptosis in a dose-dependent manner Conclusions: These data indicate that activated [ELs are the source of IL-15, which might, in turn, induce IEL prolitemtion and cytotoxic activity against enterocyles, thus leading to mucosal damage in active CD, In addition, IL-15 might favour the defect of IEL apoptosis in active CD, thus predisposing to the expansion of IELs with an abnormal phenotype and a restricted repertoire W1368 High Prevalence of Coeliac Disease in Psoriasis Veronica Ojetti, Jose Aquilar Sancbez, Cristina Guemero, Barbara Fossati, Ascanio Tridente, Rodolfo Capizzi, Messio Migneco, Paolo Amerio, Giovanni Gasbarrini, Antonio Gasbarrini Background: Diagnosis of coeliac disease (CD) may be delayed because its clinical presentation is often oligosFnptomatic. Epidemiological studies using serological tests show the disease to be much more common than previously realized Many autolmmnne disorders such as dermatitis herpetiibrmis, tyroidins, and diabetes mellitus are associated with CD. Recent AGA Abstracts A-656