A812 AGA ABSTRACTS GASTROENTEROLOGY, Vol. 108, No. 4

REACTIVE OXYGEN SPECIES (ROSI ACTIVITY AND DAMAGE IN LYMPHOCYTIC GASTRITIS (LG). I.M.Drake, D.Warland, N.Carswell, C.J.Schorab, N.Mapstone, A.T.R.Axon, M.F.Dixon, K.L.M.White. Centre for Digestive Diseases, The General Infirmary at Leeds, UK

Puruose ROS have been implicated as a cause of tissue damage in duodenal ulcer disease and H.pylori associated gastritis. We measured ROS activity and damage in LG to determine if they might also play a role in this poorly- understood condition.

Methods Subjects were recruited from patienes undergoing endoscopy for dyspepsia At endoscopy two antral and two corpus biopsies were taken for histological examination, and a further two antral biopsies for chemiluminescence (CL) (a measure of ROS activity) and malondialdehyde concentrations (MDA) (a measure of ROS mediated lipid damage).

Results Medians and interquartile ranges were:-

Normal (n=37) LG (n=7)

CL (cpm/mg) 1249 16880 467-3421 1530-39178

MDA (nmol/g) 77.5 63.3-93.1

128.5 98.4-140.6

Conclusions CL and MDA are both higher in the antral mucosa of patients with LG compared to those with normal histology (p<0.05 and p<0.01 respectively - Mann Whitney test). Neutrophils and macrophages are usually considered the chief source of ROS in inflammatory conditions, but our data suggest that lymphocytes may also be important. This work also suggests that ROS may cause tissue damage in LG, at least to lipids.

• ASSOCIATION BETWEEN ULCERATIVE COLITIS AND A POLYMORPHISM IN INTRON 2 OF THE INTERLEUKIN-1 RECEPTOR ANTAGONIST GENE. R.H. Duerr, T. Tran. Department of Medicine, University of Pittsburgh School of Medicine, Pittsburgh, PA.

Background/Aims: Insufficient production of interleukin-1 (IL-I) receptor antagonist (IL-lra) to counterbalance the observed increase in IL-I production in inflammatory bowel disease (IBD), resulting in a decreased lL-lraflL-I ratio, has been proposed as a possible mechanism in the pathogenesis of IBD. A recent report suggests that ulcerative colitis (UC) is associated with a significantly increased frequency of allele 2 of an 86-base pair (bp) variable number of tandem repeats (VNTR) polymorphism in intron 2 of the IL-lra gene. Serum perinuciear antl-neutrophil cytoplasmic antibodies (p-ANCA) have been proposed as a marker of genetic heterogeneity in UC. The purpose of this study was to determine whether UC or subgroups of UC defined by p-ANCA are associated with alleles of the 86-bp VNTR polymorphism in intron 2 of the IL-lra gene. Methods: Unrelated patients with a history of UC extending beyond the rectum (n = 94) and controls (n = 150) were studied. All study subjects were whites of European descent. The UC and control groups were well matched for Jewish ethnicity. Genomic DNA extracted from peripheral blood leukocytes was amplified in a polymerase chain reaction (PCR) using primers flanking the 86-bp VNTR polymorphic region in intron 2 of the IL-lra gene. The PCR products were resolved in an agarose gel stained with ethidium bromide. Allele assignments were made according to the sizes of the PCR products (allele 1 = 410 bp, allele 2 = 240 bp, allele 3 = 500 bp). Results for the study groups were compared using the Z 2 method and odds ratio (OR) with 95% confidence intervals (CI). Results:

Genot~'pe Study group Alleles 1,1 Alleles 1,2 Alleles 1,3 Alleles 2,2 Alleles 2,3 UC (n = 94) 44 39 3 8 0 p-ANCA+ UC (n = 50) 22 18 3 7 0 p-ANCA- UC (n = 44) 22 21 0 1 0 Controls (n = 150) 99 36 3 11 1 The frequency of at least one allele 2 was significantly increased in UC (P = .0050; OR = 2.125, CI [1.250, 3.611 ]), p-ANCA positive UC (P = .0220; OR = 2.125, CI [1.107, 4.078]), and p-ANCA negative UC (P = .0288; OR = 2.125, CI [1.073, 4.208]) compared to controls. The frequency of at least one allele 2 was identical in the p-ANCA positive and p-ANCA negative UC subgroups. The increased frequency of allele 2 in UC could be primarily attributed to an increased frequency of the alleles 1,2 genotype (P = .0040; OR = 2.245, CI [1.288, 3.914]) and a decreased frequency of the alleles 1,1 genotype (P = .0031; OR = .45, CI [.2675, .7683]). Conclusions: UC is associated with allele 2 of an 86-bp VNTR polym0rphism in intron 2 of the IL-ra gene. p-ANCA does not distinguish a subgroup of UC that is more likely to be associated with allele 2 of the 86-bp VNTR polymorphism.

• HOMOZYGOSITY FOR AN HLA CLASS II GROUP HAPLOTYPE IS ASSOCIATED WITH P-ANCA POSITIVE AND FAMILIAL ULCERATIVE COLITIS. R.H. Duerr*, LJ. Chensny*, C.T. Wongt, D.A.

* • t Neigut t . Departments of Medicine and Pediatrics, University of Pittsburgh Sebool of Medicine, Pittsburgh, PA. '

Backerunnd/Aims: HLA-DR2 is associated with Japanese ulcerative colitis (UC); but has not been consistently associated with UC in other populations. Five published, non-Japanese studies found an increased, albeit not statistically different, HLA-DR5 frequency in UC: Perinuclear anti-neutrophil cytoplasmic antibodies (p-ANCA) are found in the majorityof patients with UC and may be a marker for a more genetically homogeneous subgroup of UC. The purpose of this study was to determine whether the HLA-DR5 alleles are associated with UC or p- ANCA positive and familial UC subgroups; Methods: Unrelated patients with a history of UC extending beyond the rectum (n = 97) ted controls (n = 149) were studied. All StUdy subjects were whites of European descent.' The UC and control groups were well matched for Jewish ethnicity and gender, p-ANCA status was determined in an indirect immunofluorescence assay. UC study subjects were questioned regarding a family history of UC. HLA class II alleles were determined by polymerase chain reaction amplification of I-ILA class II sequences followed by sequence-specific oligonncleotide probe genotyping. Results for the UC study groups were compared to results for controls using the Fisher's exact test and exact estimation, of the common odds ratio (OR) with exact 95% confidence intervals (CI). Results: At least one HLA-DR5 (HLA-DRBI* 11 or -DRBI*12) allele was found in 40/149 controls, 29/97 UC patients, .18/52 p-ANCA positive UC patients, and 8/21 UC patients with a family history of UC, but the increased HLA=DR5 frequencies in the UC study groups were not statistically different than the frequency in controls. However, homozygosity for the HLA-DR5 alleles was found in 3/149 controls, 8/97 UC patients (P = .0275; OR = 4.348, C1 [1.010, 26.11]), 7/52 p-ANCA positive UC patients (P = .0034; OR = 7.475, CI [1.625, 46.63]), and 5t21 UC patients with a family history of UC (P = .0008; OR = 14.73, CI [2.598, 103.8]). The frequency of HLA-DR5 homozygosity was increased in the familial UC subgroup compared to patients with no family history ofUC (P = .0110; OR= 7.382, CI [1.289, 52.48]), and the frequency in patients with no family history of UC (3/76) was not statistically different than the frequency in co, ntrols. HLA-DR5 homozygosity was always associated w i t h homozygosity for HLA-DRB3 0202, -DQAI 050t; and ,DQBI 0301, which are in linkage disequilibrium with the HLA-DR5 alleles. Homozygosity for HLA- DRB3*0202;-DQAI*0501, or -DQBI*0301 was not as strongly associated with UC as homozygosity for the HLA,DR5 alleles, since HLA-DRB3*0202, -DQAI*0501, and -DQB1 0301 are also associated with some non-DR5 alleles. Conclusions: Homozygosity for the HLA-DR5 (HLA-DRBI*I 1 or =DRBI*I2), -DRB3*0202, -DQAI*0501, -DQBI*0301 group haplotype is associated with UC, particularly p-ANCA positive and familial UC.

E V I D E N C E S A G A I N S T M.PARATUBERCULOSIS IN C R O H N ' S D I S E A S E T I S S U E S B A S E D O N T H E

P O L Y M E R A S E C H A I N R E A C T I O N . L M. Dumonceaul ,2 ;

A. Van Gossuml ; M. Ad le r l ; J.P. Van Vooren; 1 M. Cremer l and F.

Portaels 2. (1) Department o f Gastroenterology, Erasme Universi ty Hospital, Brussels and (2) Institute o f Tropical Medicine, Antwerp, Belginm. BACKGROUND. Mycobacteria, part icularly M.paratuberculosis, have long been implicated in Crohn's disease (CD). However , detection of M.paratuberculosis in intest inal t issues by polymerase chain reaction (PCR) yie lded divergent results in independent laboratories with posi t ivi ty rates ranging from 0 to 72.5% in CD: Methodology could account for part o f these discrepancies. The aim o f this study was to test the hypothesis that mycobacter ia l infection may be involved in CD using procedures previously proven to be effect ive. METHODS. Intestinal biopsies of 72 patients (CD: N=36, ulcerative colitis: N=13, normal controls: N=23) were analyzed b y PCR for the presence of mycobacter ia at the genus level and M.paratuberculosis. A possible PCR inhibi t ion was detected by spiking template DNA : wi th the equivalent o f 2 M.para tubercu los i s genomes. RESULTS. Mycobacter ia were detected in 36 out o f 72 (50%) samples with no significant difference be tween different groups. No • M.paratuberculosis were detected in any sample. The detection l imits were 5000 and 50 mycobac t eda per sample for mycobacter ia at the genus level and M.paratuberculosis respectively. CONCLUSION. Myeobacter ia are f requent ly present in the intestinal t issues of inf lammatory bowel disease and control patients. Our data do not support any role for M.paratuberculosis in CD.