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T lymphocyte subsets in gastric tissue are altered in the presence of Helicobacter pylori
A800
AGA ABSTRACTS
EVIDENCE FOR A SPECIFIC ANTIBODY RESPONSE TO SACCHAROMTCES CEREVISIAE OLIGOMANNOSIDIC EPITOPES IN CROHN'S DISEASE. J.F.Colombel (1), B.Scndid (2), P.M.Jaequinot (3), A.Cortot (1), D.Camus (2,3), D.Poulain (2,3). Service d'H6pato-gastroent6rologie CHRU Lille (1); Laboratoire de Parasitologie-Mycologie, Facult6 de M6decine Lille (2); INSERM U42, Lille, France (3). Background/Alms: Elevated antibody levels against the yeast Saccharomyces eerevisiae have been reported in sera of patients with Crohn's disease (CD) and not ulcerative colitis (UC). The aim of this study was to identify the nature of the epitopes stimulating this antibody response. Methods: Whole cells from different S. cerevisiae strains were selected in immunofluoreseance assays for their ability to differentiate the antibody responses of CD and UC patients. Their cell wall phosphopeptidomannans (PPM) were then tested as antigen in ELISA against sera from 42 patients with CD, 20 patients with UC and 34 healthy controls. Graded chemical degradations were then performed on the most reactive strain. The domain of the molecule bearing the discriminating epitopes was determined through gas-liquid chromatography-mass spectrometry. Results: The greatest discrimination between patients with CD and UC was obtained with Saccharomyces uvarum 1 (Sul), a S. cerevisiae strain used in the brewing of beer. ELISA directed against Sul PPM was 64% ' sensitive and 77% specific discriminating CD vs UC and 71% sensitive and 89% specific for CD vs controls. Periodic oxidation and selective degradation demonstrated that the polysaccharide epitope of importance was shared by both the acid stable and alkali labile domains of the PPM. Determination of oligomannose sequences of Sul PPM suggests that a mannotetrose Man(I--,.3) Man(I-,2) Man(I---,2) Man was responsible for the serological response seen in CD. Conclusion: An antibody response against a sequence of mannose residues was specifically associated with CD. Further identification of this immunogen may help to understand its potential as a disease marker and why this response occurs at all.
• SERUM TRANSFERRIN RECEPTOR/FERHITIN RATIO IS A RELIABLE PREDICTOR OF IRON DEFICIENCY ANEMIA IN INFLAMMATORY BOWEL DISEASE PATIENTS. C. c o l t o n r K. Geraci, H. Tahsildar, C. Flowers, J. Cook, G. Cooper, K. Hornbuckle. Depts. of Medicine, Case Western Reserve Univeristy, Cleveland, OH and Kansas University, Kansas City, Kansas. Background~Aims: Iron studies are d i f f i c u l t to interpret in chronic disease states such as inflammatory bowel disease(IBD). Recently, the serum Transferrin Receptor/Ferritin ratio(TRF) has b e e n r e p o r t e d t o be a r e l i a b l e i n d i c a t o r of iron deficiency. The aim of this study is to d e t e r m i n e if the TRF is a r e l i a b l e p r e d i c t o r Of iron d e f i c i e n c y in IBD. Methods: 18 c o n s e c u t i v e IBD patients with anemia(Hct.<35%) were asked to p a r t i c i p a t e ; 16 consented. Two p a t i e n t s were excluded(serum lost in one patient, hemolysis in the second patient), l e a v i n g 14 participants. There were 8 p a t i e n t s w i t h u l c e r a t i v e colitis(UC) and 6 w i t h Crohn's~ disease(CD); 4 w e r e male and 10 were females. The m e a n age was 52. Serum iron, t r a n s f e r r i n saturation, ferritin, t r a n s f e r r i n receptor level(TR) Via ELISA, and bone marrow biopsy were obtained from each. The TRF was calculated by the formula: (TR/ferritin)x1000. Activity of d i s e a s e was d e t e r m i n e d by flexible sigmoidoscopy in UC and by the H a r v e y - B r a d s h a w index in CD. Results: By bone m a r r o w b i o p s y 10 p a t i e n t s had no iron stores , and 4 p a t i e n t s had e l e v a t e d iron stores. The m e a n T F R was 53.1 in those with elevated iron stores and 1314.0 in iron deficient patients(p=0.009 by Wilc0~on analysis). Positive Negative PredictivePredictive Sens. Spee. Value Value Ferritin(<15u~/dl) 50% 100% 100% 33% TRF(>I00) 90% 100% 100% 80% Conclusions: The TRF is a r e l i a b l e p r e d i c t o r of iron deficiency in IBD patients, and is more s e n s i t i v e than serum f e r r i t i n in these patients.
GASTROENTEROLOGY, VoI. IO8, No. 4
• HLA CLASS H GENES AND CROHN'S DISEASE : DIFFERENT ASSOCIATIONS IN ADULTS AND CHILDREN. JF Colombel. PM Danz6, S Jacquot, MN Loste, D Heresbach, S Ategbo, B P6richon, G Semana, JP C6Zard. CHRU Lille, St-Louis-Paris, R.Debr6-Paris et Rennes; CRTS Rannes et INSERM U120, Paris, France. Available studies of association between HLA class II genes and Crohn's disease (CD) have given various results. Those discrepancies might be partly due to heterogeneity in age of studied patients. The aim of the study was to measure frequencies of HLA class II genes among two different populations of patients with CD diagnosed when adult (>16 years) or during childhood (< 16 years). Methods : 258 adults (137 F, 121 H, mean age 22 years) and 75 children with CD (35F, 40H, mean age 11.5 years) had molecular genotyping of HLADQA1, DQB1 and DRB1 alleles using molecular biology (PCR-SSO or PCR'RFLP). The results were compared in an ethnically matched control population of 486 caucasian adults using Chi2 test with p corrected value. Results : Different genes frequencies in the 3 groups (in %) : Adults with CD Children with CD Controls ~n = 258) (n= 75) ~n = 486) DQB1*0602.3 12.8%* 16.2% 19% 14.7% 19.7%* 10.2% DOBI*0501 DRB1*01 14.6%$ 16.2%$ 9% 13.3% 6.5%* 5.6%$ DRBI*03 DRBI*07 17.6%§ 12.7% 11.1% * : p<0.025 vs Controls; $ : p<0.01 vs Controls; § : p<0.001 vs Controls. Conclusion : There was a positiveassociation between DRBI*01 gane and a negative association between DRB1*03 gene and CD in both adults and children. Different associations observed between DQB1 and DRBI*07 genes in children and adults suggest that those genes may influenCe the age of onset of CD.
T L Y M P H O C Y T E S U B S E T S IN G A S T R I C T I S S U E A R E A L T E R E D IN T H E P R E S E N C E OF HELICOBACTER PYLORI. C.Colton t S.Czinn, S.Setrakian, M.Sy, A.Chak, K.Ferguson. Depts. of Medicine, Pediatrics and Pathology. Case W e s t e r n R e s e r v e University, Cleveland; OH. Background/Aims: The p a t h o g e n e s i s of inflammation in tissue infected b y H. pylorl has not been completely elucidated. The aim of this study is to determine T helper(CD4) and T cytolytic(CD8) lymphocyte density and ratios in gastric tissue infected by H. py!ori as compared to control tissue. Methods: Antral biopsies were o b t a i n e d from patients u n d e r g o i n g routine endoscopy. The presence of H. pylori was d e t e r m i n e d by H e m a t o x y l i n and Eosin and G i e m s a s t a i n s . The lamina p r o p r i a of frozen tissue sections were examined using an i m m u n o h i s t o c h e m i c a l assay, labelled streptavidin-biotin(LSAB), to visualize CD4 and CD8 m o l e c u l e s microscopically. Results: B i o p s y specimens were obtained from 20 patients. Tissue sections from 3 controls w i t h normal tissue histol o g i c a l l y had a m e a n of 2.3 CD4 and 2.6 CD8 cells per high power field(HPF) by LSAB. 10 tissue sections with chronic inflammation without H . pylori had a m e a n of 18.2 CD4 and 12.1 CD8 cells per HPF. M e a n CD4:CD8 ratio for these patients was 1.53(range 0.3-2.5). For the 7 patient~ with chronic inflammation and ~. pylori present the mean CD4 and CD0 ce!l count~ per EPF were 13.2 and 17.9 respectively. Mean CD4:CD3 ratio for these patients was 0.82(range 0.1-0.9). The difference between CD4:CD8 ratios for the two groups with chronic i n f l a m m a t i o n was s t a t i s t i c a l l y significant by Wilcoxon analysis(p=0.03). Conclusions: CD4 and CD8 cells p l a y a role in the i n f l a m m a t o r y response of H. pylori infected tissue, and t h e i r r a t i o as compared to inflammed tissue without H. pylori is reversed. This suggests a more prominent role of CD8 lymphocytes in H. pylori infected tissue as compared to i n f l a m m e d control tissue.
AGA ABSTRACTS
EVIDENCE FOR A SPECIFIC ANTIBODY RESPONSE TO SACCHAROMTCES CEREVISIAE OLIGOMANNOSIDIC EPITOPES IN CROHN'S DISEASE. J.F.Colombel (1), B.Scndid (2), P.M.Jaequinot (3), A.Cortot (1), D.Camus (2,3), D.Poulain (2,3). Service d'H6pato-gastroent6rologie CHRU Lille (1); Laboratoire de Parasitologie-Mycologie, Facult6 de M6decine Lille (2); INSERM U42, Lille, France (3). Background/Alms: Elevated antibody levels against the yeast Saccharomyces eerevisiae have been reported in sera of patients with Crohn's disease (CD) and not ulcerative colitis (UC). The aim of this study was to identify the nature of the epitopes stimulating this antibody response. Methods: Whole cells from different S. cerevisiae strains were selected in immunofluoreseance assays for their ability to differentiate the antibody responses of CD and UC patients. Their cell wall phosphopeptidomannans (PPM) were then tested as antigen in ELISA against sera from 42 patients with CD, 20 patients with UC and 34 healthy controls. Graded chemical degradations were then performed on the most reactive strain. The domain of the molecule bearing the discriminating epitopes was determined through gas-liquid chromatography-mass spectrometry. Results: The greatest discrimination between patients with CD and UC was obtained with Saccharomyces uvarum 1 (Sul), a S. cerevisiae strain used in the brewing of beer. ELISA directed against Sul PPM was 64% ' sensitive and 77% specific discriminating CD vs UC and 71% sensitive and 89% specific for CD vs controls. Periodic oxidation and selective degradation demonstrated that the polysaccharide epitope of importance was shared by both the acid stable and alkali labile domains of the PPM. Determination of oligomannose sequences of Sul PPM suggests that a mannotetrose Man(I--,.3) Man(I-,2) Man(I---,2) Man was responsible for the serological response seen in CD. Conclusion: An antibody response against a sequence of mannose residues was specifically associated with CD. Further identification of this immunogen may help to understand its potential as a disease marker and why this response occurs at all.
• SERUM TRANSFERRIN RECEPTOR/FERHITIN RATIO IS A RELIABLE PREDICTOR OF IRON DEFICIENCY ANEMIA IN INFLAMMATORY BOWEL DISEASE PATIENTS. C. c o l t o n r K. Geraci, H. Tahsildar, C. Flowers, J. Cook, G. Cooper, K. Hornbuckle. Depts. of Medicine, Case Western Reserve Univeristy, Cleveland, OH and Kansas University, Kansas City, Kansas. Background~Aims: Iron studies are d i f f i c u l t to interpret in chronic disease states such as inflammatory bowel disease(IBD). Recently, the serum Transferrin Receptor/Ferritin ratio(TRF) has b e e n r e p o r t e d t o be a r e l i a b l e i n d i c a t o r of iron deficiency. The aim of this study is to d e t e r m i n e if the TRF is a r e l i a b l e p r e d i c t o r Of iron d e f i c i e n c y in IBD. Methods: 18 c o n s e c u t i v e IBD patients with anemia(Hct.<35%) were asked to p a r t i c i p a t e ; 16 consented. Two p a t i e n t s were excluded(serum lost in one patient, hemolysis in the second patient), l e a v i n g 14 participants. There were 8 p a t i e n t s w i t h u l c e r a t i v e colitis(UC) and 6 w i t h Crohn's~ disease(CD); 4 w e r e male and 10 were females. The m e a n age was 52. Serum iron, t r a n s f e r r i n saturation, ferritin, t r a n s f e r r i n receptor level(TR) Via ELISA, and bone marrow biopsy were obtained from each. The TRF was calculated by the formula: (TR/ferritin)x1000. Activity of d i s e a s e was d e t e r m i n e d by flexible sigmoidoscopy in UC and by the H a r v e y - B r a d s h a w index in CD. Results: By bone m a r r o w b i o p s y 10 p a t i e n t s had no iron stores , and 4 p a t i e n t s had e l e v a t e d iron stores. The m e a n T F R was 53.1 in those with elevated iron stores and 1314.0 in iron deficient patients(p=0.009 by Wilc0~on analysis). Positive Negative PredictivePredictive Sens. Spee. Value Value Ferritin(<15u~/dl) 50% 100% 100% 33% TRF(>I00) 90% 100% 100% 80% Conclusions: The TRF is a r e l i a b l e p r e d i c t o r of iron deficiency in IBD patients, and is more s e n s i t i v e than serum f e r r i t i n in these patients.
GASTROENTEROLOGY, VoI. IO8, No. 4
• HLA CLASS H GENES AND CROHN'S DISEASE : DIFFERENT ASSOCIATIONS IN ADULTS AND CHILDREN. JF Colombel. PM Danz6, S Jacquot, MN Loste, D Heresbach, S Ategbo, B P6richon, G Semana, JP C6Zard. CHRU Lille, St-Louis-Paris, R.Debr6-Paris et Rennes; CRTS Rannes et INSERM U120, Paris, France. Available studies of association between HLA class II genes and Crohn's disease (CD) have given various results. Those discrepancies might be partly due to heterogeneity in age of studied patients. The aim of the study was to measure frequencies of HLA class II genes among two different populations of patients with CD diagnosed when adult (>16 years) or during childhood (< 16 years). Methods : 258 adults (137 F, 121 H, mean age 22 years) and 75 children with CD (35F, 40H, mean age 11.5 years) had molecular genotyping of HLADQA1, DQB1 and DRB1 alleles using molecular biology (PCR-SSO or PCR'RFLP). The results were compared in an ethnically matched control population of 486 caucasian adults using Chi2 test with p corrected value. Results : Different genes frequencies in the 3 groups (in %) : Adults with CD Children with CD Controls ~n = 258) (n= 75) ~n = 486) DQB1*0602.3 12.8%* 16.2% 19% 14.7% 19.7%* 10.2% DOBI*0501 DRB1*01 14.6%$ 16.2%$ 9% 13.3% 6.5%* 5.6%$ DRBI*03 DRBI*07 17.6%§ 12.7% 11.1% * : p<0.025 vs Controls; $ : p<0.01 vs Controls; § : p<0.001 vs Controls. Conclusion : There was a positiveassociation between DRBI*01 gane and a negative association between DRB1*03 gene and CD in both adults and children. Different associations observed between DQB1 and DRBI*07 genes in children and adults suggest that those genes may influenCe the age of onset of CD.
T L Y M P H O C Y T E S U B S E T S IN G A S T R I C T I S S U E A R E A L T E R E D IN T H E P R E S E N C E OF HELICOBACTER PYLORI. C.Colton t S.Czinn, S.Setrakian, M.Sy, A.Chak, K.Ferguson. Depts. of Medicine, Pediatrics and Pathology. Case W e s t e r n R e s e r v e University, Cleveland; OH. Background/Aims: The p a t h o g e n e s i s of inflammation in tissue infected b y H. pylorl has not been completely elucidated. The aim of this study is to determine T helper(CD4) and T cytolytic(CD8) lymphocyte density and ratios in gastric tissue infected by H. py!ori as compared to control tissue. Methods: Antral biopsies were o b t a i n e d from patients u n d e r g o i n g routine endoscopy. The presence of H. pylori was d e t e r m i n e d by H e m a t o x y l i n and Eosin and G i e m s a s t a i n s . The lamina p r o p r i a of frozen tissue sections were examined using an i m m u n o h i s t o c h e m i c a l assay, labelled streptavidin-biotin(LSAB), to visualize CD4 and CD8 m o l e c u l e s microscopically. Results: B i o p s y specimens were obtained from 20 patients. Tissue sections from 3 controls w i t h normal tissue histol o g i c a l l y had a m e a n of 2.3 CD4 and 2.6 CD8 cells per high power field(HPF) by LSAB. 10 tissue sections with chronic inflammation without H . pylori had a m e a n of 18.2 CD4 and 12.1 CD8 cells per HPF. M e a n CD4:CD8 ratio for these patients was 1.53(range 0.3-2.5). For the 7 patient~ with chronic inflammation and ~. pylori present the mean CD4 and CD0 ce!l count~ per EPF were 13.2 and 17.9 respectively. Mean CD4:CD3 ratio for these patients was 0.82(range 0.1-0.9). The difference between CD4:CD8 ratios for the two groups with chronic i n f l a m m a t i o n was s t a t i s t i c a l l y significant by Wilcoxon analysis(p=0.03). Conclusions: CD4 and CD8 cells p l a y a role in the i n f l a m m a t o r y response of H. pylori infected tissue, and t h e i r r a t i o as compared to inflammed tissue without H. pylori is reversed. This suggests a more prominent role of CD8 lymphocytes in H. pylori infected tissue as compared to i n f l a m m e d control tissue.