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Celiac disease associated with HLA-DQ8 and DQ2 have different T-cell repertoires in vivo
(1) Maxton et al Gut 1991; 332: 662; (2) Sullivan et al N Z Med J 1994; ]07: 428; (3) Lewiset al Gut 2001; 48:216
were detected for bone mineral densities measured at other levels. Clinical parameters at diagnosis did not evidenced differences comparing non-carriers with carrier patients. CONCLUSIONS: Our study evidenced, for the first time, that a genetic susceptibility is associated with low bone mass of CD patients. Carriers of the allele 2 of the IL1B-511 of IL 1B gene had significantlylower bone density at the total skeleton level than non-carriers. CD osteopathy is the result of a combination of factors such as local and systemic immunological disturbances, malabsorptive nutritional factors. Our study suggests that genes controlling the regulation if the immune response are also involved.
126 Segmental Gas Transit in Patients with Abdominal Bloating 8eatnce Salvioli, Jordi Serra, Carlos Lorenzo, Santiago Aguade, Femando Azpiroz, Jnan-R. Malagelada Usinga gas challenge test we have previously shown that patients complaining of abdominal bloating exhibit impaired gas tolerance and develop retention. Our present aim was to identify the gut compartment responsible for gas retention in these patients. METHODS. In 12 IBS patients with abdominal bloating, and 10 healthy subjects, gut transit of radiolabelled gas was measured by scintigraphy, as follows. A gas nuxture (NI, CO2, Ox in venous proportions) was infused at 24 ml/min into the jejunum for 2 h, while gas outflow was collected via an mtrarectal cannula. After 1 h, a 74 mBq bolus of t33Xe was added to the gasmixture, and anterior and posterior scans were simultaneously obtained by means of a largeheld-of-view dual head gammacamera at 60 s intervals for the following hour. Segmental gas tramit was measured by a region of interest program. At the end of the study we measured gas retention, as gas volume infused minus evacuated, and perception by a 0-6 scale.RESULTS. [n patients total gut transit of gas was delayed (T~0 42 +/-4 rain vs 31 +/3 rnm in health; p<0.05) and this was associated to gas retention (589+A194 ml by the end of the study vs -72+A18 ml in health; p<0.05) and abdominal symptoms (score 3.5+/-0.6 vs 1.3 +/-0.3 in health; p<0.05). Segmental transit analysis showed that impaired gasclearance was related to delayed gas transit through the small intestine (Tso 21 +/-3 min vs 1).+/-3 min in health; p<0,05), whereas colonic transit was normal (T~0 20+/-2 min and 19+/-2 rain in health; N.S.). No gaseous back flow was detected in any study. CONCLUSION. Small bowel incompetence to propulse gas is responsible for gas intolerance and retention in patients complaining of abdominal bloating
129 Anti-Neuronal Antibodies in Patients with Celiac Disease and Neurological Disorders Umberto Volta, Roberto De Giorgio, Nunzio Petrofini, Vincenao Stanghellini, Giovanni Barbara, Alessandro Granito, Lorenza Veronesi, Fabrizio De Ponti, Elisa Fustini, Roberto Corinaldesi, Francesco Bianchi Neurological disorders have been reported in patients with celiac disease (CD). However, the clinical and immunological features of CD presenting with neurological dysfunction have been scarcely investigated. Aims of this study were to: 1) assess the prevalence of neurological disorders in CD; 2) define the clinical features of this subset of caliacs; 3) evaluate the effect of ghiten-free diet (GFD) on the clinical course of their neurological manifestations; and 4) analyze the i~revalence of anti-neuronal antibodies (NA) in CD. Methods: Neurological symptoms were investigated in 160 celiacs (120 women; median age: 37 yrs). NA to central/enteric nervous systems were investigated in all celiacs with neurological disorders, in 20 celiacs without neurological dysfunction and in 30 controls using indirect immunofluorescence on rat and primate cerebellar cortex and rat intestmal sections. Results: Thirteen (8%) of the 160 patients had neurological disorders including 3 cases of epilepsy, 3 with attention/memory impairment, 2 with cerebellar ataxia, 2 with peripheral neuropathy, 1 with multiple sclerosis, 1 with acute cerebrovascular dysfunction and 1 with myotonic dystrophy. In all but 2 of the 13 cases, the neurological disorder preceded diagnosis of CD. No significant clinical difference was found between celiacs with and without neurological involvement since both groups showed a similar median age, a higher prevalence of female gender and absence of gastrointestinal signs in about a half cases. Neurological symptoms improved or disappeared in 7 patients who started a GFD within 6 momhs after neurologica! onset, and in none of 4 patients who began later. The prevalence of central-nervous-system NA was significantly higher in neurological cefiacs (61%) than in other patients (5%) (P<0.001) or controls (0%) (P
127 Adrenergicand Serotonergic Polymorphisms: Association with Symptom Phenotype and Psychosomatic Scores in Lower Functional Bowel Disorders H. Jae Kim, Michael Camilleri, Paula J. Carlson, Filippo Cremonini, Irene Ferber, Debra Stephens, Sanna McKinzie, Alan R. Zinsmeister, Paul Urrutia Background:Adrenergic and serotonergic mechanisms influence colonic functions in irritable bowelsyndrome (IBS). Psychological disturbances and IBS are partly genetically determined. Hypothesis:Alpha-2 adrenoreceptors (alpha-2 AR), norepinephrine transporter (NET), and ser0t0nintransporter promoter (SERT-P) polymorphisms are associated with IBS and psychosomatic (PSC) phenotypes. Aims: 1) To estimate prevalence of alpha-2 AR (Ncil, Mspl, and Styl), SERT-P, and NET polymorphisms in 276 patients with IBS or chronic abdominal pain(CAP) and 120 ethnically similar controls. 2) To assess the association of the polymorphlsmswith IBS subgroups an~i CAP. Methods: A validated bowel disease and PSC questionnaire characterized the IBS phenotype and CAP. Candidate genes were based on known associationsof adrenergic and serotonergic genotypes with smooth muscle or psychological dtsorders.PoIymorphisms were idemified by PCR-based restriction fragment length polymorphisms,and confirmed by direct sequencing. A logistic regression analysis was used to assess the association of different polymorphisms : Alpha-2 AR (ww: wild type, wp: heterozygons, pp:polymorphic homozyguus) or SERT-P (ll and ss: long and short homozygons, Is: beterozygoas)wltli InS or CAP phenotypes. A multiple (linear) regression analysis was used to assess the association of PSC score with the different polymorphisms. Results: Sty1 and NET polymorphisms were not identified in the first 100 patients and were not measured in remainingsubjects. Distributions of Ncil and Mspl, and SERT-P polymorphisms were not associated overall with lower FBD vs. controls. Presence of either Ncil (wp/pp) or Mspl (wp/pp)polymorphism was associated with an increased odds for the Ins constipation (IBSC) phenotype. For wp/pp relative to ww genotypes, the odds ratio and (95%CI) for IBS-C vs. controlswas 2.48 (0.98-6.28), p = 0.05 using Ncil and 1.66 (0.94-2.92), p ~ 0.08 asmgMsp1. Combination of Ncil (wp/pp) and SERT-P (ss/ls) polymorphisms was associated ~lth higher PSC scores (p <0.05) in lower functional bowel disorders, suggesting that these combined genotypes explain in part the variation in PSC scores amongst different FBD phenotypes. Conclusion: The observed association in lower functional bowel disorders of specific alpha-2 AR genotypes with 1BS-C phenotype, and of combined alpha-2 AR and SERT-Pgenotypes with PSC scores warrants further study.
130 Celiac Disease Associated with HLA-DQ8 and DQ2 Have Different T-Cell Repertoires in Vivo Robert P. Anderson, David Van Heel, Arthur Tatham, Martin Bamardo, Derek P. Jewefl, Adrian V. Hill Celiac disease (CD) is associated with HLA-DQ2 (>90%) or HLA-DQ8 (5%). HLA-DQ2 and DQ8-resticted gluten epitopes for intestinal T-cell clones are known, but their relevance in vivo is uncertain. Definition of peptides targeted by gluten-specific T-cells in vivo may allow developmem of immunotherapy, but peptides targeted in HLA-DQ8-associated CD may differ from those in DQ2 CD. AIM: Define the hierarchy of wheat gliadin T-cell epitopes induced by gluten challenge in HLA-DQ8 + CD. METHODS: CD patients were identified as DQ2 + 84 DQ2 + 8 +, or DQ8 + 2- by genotyping HLA-DQAand HLA-DQBalleles. Three DQ8+2- and 6 D Q 2 + 8 + CD and 2 DQ8+2- healthy subjects following gluten free diet (GFD) for > 4 weeks had 4 slices gluten bread daily for 3 days, and blood was collected on day 0, and/or 6 6z 7 (as described in Anderson RP et al Nature Medicine 6:337). Interferon gamma ([FN~) ELlspot assays were performed using PBMC incubated overnight with a panel of 652 overlapping 20mers spanning all gene-derived Tricitum aestivum call3- (61), ~- (47) and co-gliadin (3) sequences in Genbank. RESULTS: There were no peptide-specific responses in healthy subjects before or after gluten challenge. In 2/3 HLA-DQ8 + (not 2) CD subjects, IFN'y ELIspot responses were induced after gluten challenge specific for deamidated gliadin. There were clearly dominant responses for specific, previously un-reported cdlBgliadin peptides (different in each subject). Responses to these dominant peptides were enhanced by deamidation. Repeat gluten challenge in one subject allowed mapping of the optimal sequence and deamidation site in the dominant epitope, lmmunomagnetic bead depletion of PBMC revealed CD4 T cells expressing the gut homing integrin (a4~7) were responsible for the gliadin peptide-specific IFN'/secretion. Neither HLA-DQ8 + 2- subject had T cells induced specific for the dominant gliadin epitope induced in HLA-DQ2 + CD subjects (homologous to A-gliadin 57-73). In contrast, none of the HLA-DQ2 + 8 + subjects showed induction of ]FN'y secreting T-cells specific [or the epitopes dominant in HLADQ8 + 2 + CD subjects. In 4/6 DQ2 + 8 + subjects there was induction of T-cells specific for homologues of A-gfiadin 57-73. CONCLUSION: Epitopes of gfiadin-specific T-cells are different in HLA-DQ8 and DQ2 associated CD. In CD associated with HLA-DQ2 and DQ8, HLA-DQ2 associated epitopes dominate. Therefore, peptide immunotherapy for CD will be HLA-type specific, or include a cocktail of HLA-DQ2 and DQ8- restricted gliadin epitopes.
128 LowBone Mass in Celiac Disease ls Associated with an Allelic Variation in the ILIB Gene
MariaL. Moreno, Laura Murillo, Horacio Vazquez, Roberto M. Mazure, Alejandra Chemavsky,Alicia Sambuelfi, Edgardo Smecuol, Sonia Niveloni, Silvia Pedreira, M. De Btssio,Eduardo Maurino, Amado S. Pena, Julio C. Bai BACKGROUND/AIM: Celiac disease (CD) is frequently associated with decreased bone mineralmass and an increased rate of fractures in the peripheral skeleton. The interleukin IB(IL-1B)has been implicated in both, the intestinal lesion of CD and the bone remodeling processAn allelic variant of the gene ILl B-511 has been associated with bone demineralizatton in the general population and in patients with Crohn's disease. Our objective was to determinewhether decreased bone mass in CD patients is related with gene polymorphisms encodingfor 1L-lB. MATERIALS & METHODS: We studied 71 CD patients and 60 healthy controlsubjects. At the time of diagnosis, all patients included underwent bone densitometric studies at, either or both, the femoral neck, lumbar spine (L2-L4) and total skeleton. The IL1B-5l1 biallelic polymorphism was genotyped by PCR. RESULTS: (XS• CD patiems have a significantly greater homozygozity for the allele 2 of the [L1B-511 (52%) than that determined in the control population (25%; p<0.01). Compared with non-carriers CD patients,carriers of the allele 2 of the IL1B-511 had significantly lower bone mass (Z-score) atthe total skeleton level (-0.21 • 0.56 vs. - 1.09 - 0.40; p = 0.0488). No statistical differences
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AGA Abstracts
were detected for bone mineral densities measured at other levels. Clinical parameters at diagnosis did not evidenced differences comparing non-carriers with carrier patients. CONCLUSIONS: Our study evidenced, for the first time, that a genetic susceptibility is associated with low bone mass of CD patients. Carriers of the allele 2 of the IL1B-511 of IL 1B gene had significantlylower bone density at the total skeleton level than non-carriers. CD osteopathy is the result of a combination of factors such as local and systemic immunological disturbances, malabsorptive nutritional factors. Our study suggests that genes controlling the regulation if the immune response are also involved.
126 Segmental Gas Transit in Patients with Abdominal Bloating 8eatnce Salvioli, Jordi Serra, Carlos Lorenzo, Santiago Aguade, Femando Azpiroz, Jnan-R. Malagelada Usinga gas challenge test we have previously shown that patients complaining of abdominal bloating exhibit impaired gas tolerance and develop retention. Our present aim was to identify the gut compartment responsible for gas retention in these patients. METHODS. In 12 IBS patients with abdominal bloating, and 10 healthy subjects, gut transit of radiolabelled gas was measured by scintigraphy, as follows. A gas nuxture (NI, CO2, Ox in venous proportions) was infused at 24 ml/min into the jejunum for 2 h, while gas outflow was collected via an mtrarectal cannula. After 1 h, a 74 mBq bolus of t33Xe was added to the gasmixture, and anterior and posterior scans were simultaneously obtained by means of a largeheld-of-view dual head gammacamera at 60 s intervals for the following hour. Segmental gas tramit was measured by a region of interest program. At the end of the study we measured gas retention, as gas volume infused minus evacuated, and perception by a 0-6 scale.RESULTS. [n patients total gut transit of gas was delayed (T~0 42 +/-4 rain vs 31 +/3 rnm in health; p<0.05) and this was associated to gas retention (589+A194 ml by the end of the study vs -72+A18 ml in health; p<0.05) and abdominal symptoms (score 3.5+/-0.6 vs 1.3 +/-0.3 in health; p<0.05). Segmental transit analysis showed that impaired gasclearance was related to delayed gas transit through the small intestine (Tso 21 +/-3 min vs 1).+/-3 min in health; p<0,05), whereas colonic transit was normal (T~0 20+/-2 min and 19+/-2 rain in health; N.S.). No gaseous back flow was detected in any study. CONCLUSION. Small bowel incompetence to propulse gas is responsible for gas intolerance and retention in patients complaining of abdominal bloating
129 Anti-Neuronal Antibodies in Patients with Celiac Disease and Neurological Disorders Umberto Volta, Roberto De Giorgio, Nunzio Petrofini, Vincenao Stanghellini, Giovanni Barbara, Alessandro Granito, Lorenza Veronesi, Fabrizio De Ponti, Elisa Fustini, Roberto Corinaldesi, Francesco Bianchi Neurological disorders have been reported in patients with celiac disease (CD). However, the clinical and immunological features of CD presenting with neurological dysfunction have been scarcely investigated. Aims of this study were to: 1) assess the prevalence of neurological disorders in CD; 2) define the clinical features of this subset of caliacs; 3) evaluate the effect of ghiten-free diet (GFD) on the clinical course of their neurological manifestations; and 4) analyze the i~revalence of anti-neuronal antibodies (NA) in CD. Methods: Neurological symptoms were investigated in 160 celiacs (120 women; median age: 37 yrs). NA to central/enteric nervous systems were investigated in all celiacs with neurological disorders, in 20 celiacs without neurological dysfunction and in 30 controls using indirect immunofluorescence on rat and primate cerebellar cortex and rat intestmal sections. Results: Thirteen (8%) of the 160 patients had neurological disorders including 3 cases of epilepsy, 3 with attention/memory impairment, 2 with cerebellar ataxia, 2 with peripheral neuropathy, 1 with multiple sclerosis, 1 with acute cerebrovascular dysfunction and 1 with myotonic dystrophy. In all but 2 of the 13 cases, the neurological disorder preceded diagnosis of CD. No significant clinical difference was found between celiacs with and without neurological involvement since both groups showed a similar median age, a higher prevalence of female gender and absence of gastrointestinal signs in about a half cases. Neurological symptoms improved or disappeared in 7 patients who started a GFD within 6 momhs after neurologica! onset, and in none of 4 patients who began later. The prevalence of central-nervous-system NA was significantly higher in neurological cefiacs (61%) than in other patients (5%) (P<0.001) or controls (0%) (P
127 Adrenergicand Serotonergic Polymorphisms: Association with Symptom Phenotype and Psychosomatic Scores in Lower Functional Bowel Disorders H. Jae Kim, Michael Camilleri, Paula J. Carlson, Filippo Cremonini, Irene Ferber, Debra Stephens, Sanna McKinzie, Alan R. Zinsmeister, Paul Urrutia Background:Adrenergic and serotonergic mechanisms influence colonic functions in irritable bowelsyndrome (IBS). Psychological disturbances and IBS are partly genetically determined. Hypothesis:Alpha-2 adrenoreceptors (alpha-2 AR), norepinephrine transporter (NET), and ser0t0nintransporter promoter (SERT-P) polymorphisms are associated with IBS and psychosomatic (PSC) phenotypes. Aims: 1) To estimate prevalence of alpha-2 AR (Ncil, Mspl, and Styl), SERT-P, and NET polymorphisms in 276 patients with IBS or chronic abdominal pain(CAP) and 120 ethnically similar controls. 2) To assess the association of the polymorphlsmswith IBS subgroups an~i CAP. Methods: A validated bowel disease and PSC questionnaire characterized the IBS phenotype and CAP. Candidate genes were based on known associationsof adrenergic and serotonergic genotypes with smooth muscle or psychological dtsorders.PoIymorphisms were idemified by PCR-based restriction fragment length polymorphisms,and confirmed by direct sequencing. A logistic regression analysis was used to assess the association of different polymorphisms : Alpha-2 AR (ww: wild type, wp: heterozygons, pp:polymorphic homozyguus) or SERT-P (ll and ss: long and short homozygons, Is: beterozygoas)wltli InS or CAP phenotypes. A multiple (linear) regression analysis was used to assess the association of PSC score with the different polymorphisms. Results: Sty1 and NET polymorphisms were not identified in the first 100 patients and were not measured in remainingsubjects. Distributions of Ncil and Mspl, and SERT-P polymorphisms were not associated overall with lower FBD vs. controls. Presence of either Ncil (wp/pp) or Mspl (wp/pp)polymorphism was associated with an increased odds for the Ins constipation (IBSC) phenotype. For wp/pp relative to ww genotypes, the odds ratio and (95%CI) for IBS-C vs. controlswas 2.48 (0.98-6.28), p = 0.05 using Ncil and 1.66 (0.94-2.92), p ~ 0.08 asmgMsp1. Combination of Ncil (wp/pp) and SERT-P (ss/ls) polymorphisms was associated ~lth higher PSC scores (p <0.05) in lower functional bowel disorders, suggesting that these combined genotypes explain in part the variation in PSC scores amongst different FBD phenotypes. Conclusion: The observed association in lower functional bowel disorders of specific alpha-2 AR genotypes with 1BS-C phenotype, and of combined alpha-2 AR and SERT-Pgenotypes with PSC scores warrants further study.
130 Celiac Disease Associated with HLA-DQ8 and DQ2 Have Different T-Cell Repertoires in Vivo Robert P. Anderson, David Van Heel, Arthur Tatham, Martin Bamardo, Derek P. Jewefl, Adrian V. Hill Celiac disease (CD) is associated with HLA-DQ2 (>90%) or HLA-DQ8 (5%). HLA-DQ2 and DQ8-resticted gluten epitopes for intestinal T-cell clones are known, but their relevance in vivo is uncertain. Definition of peptides targeted by gluten-specific T-cells in vivo may allow developmem of immunotherapy, but peptides targeted in HLA-DQ8-associated CD may differ from those in DQ2 CD. AIM: Define the hierarchy of wheat gliadin T-cell epitopes induced by gluten challenge in HLA-DQ8 + CD. METHODS: CD patients were identified as DQ2 + 84 DQ2 + 8 +, or DQ8 + 2- by genotyping HLA-DQAand HLA-DQBalleles. Three DQ8+2- and 6 D Q 2 + 8 + CD and 2 DQ8+2- healthy subjects following gluten free diet (GFD) for > 4 weeks had 4 slices gluten bread daily for 3 days, and blood was collected on day 0, and/or 6 6z 7 (as described in Anderson RP et al Nature Medicine 6:337). Interferon gamma ([FN~) ELlspot assays were performed using PBMC incubated overnight with a panel of 652 overlapping 20mers spanning all gene-derived Tricitum aestivum call3- (61), ~- (47) and co-gliadin (3) sequences in Genbank. RESULTS: There were no peptide-specific responses in healthy subjects before or after gluten challenge. In 2/3 HLA-DQ8 + (not 2) CD subjects, IFN'y ELIspot responses were induced after gluten challenge specific for deamidated gliadin. There were clearly dominant responses for specific, previously un-reported cdlBgliadin peptides (different in each subject). Responses to these dominant peptides were enhanced by deamidation. Repeat gluten challenge in one subject allowed mapping of the optimal sequence and deamidation site in the dominant epitope, lmmunomagnetic bead depletion of PBMC revealed CD4 T cells expressing the gut homing integrin (a4~7) were responsible for the gliadin peptide-specific IFN'/secretion. Neither HLA-DQ8 + 2- subject had T cells induced specific for the dominant gliadin epitope induced in HLA-DQ2 + CD subjects (homologous to A-gliadin 57-73). In contrast, none of the HLA-DQ2 + 8 + subjects showed induction of ]FN'y secreting T-cells specific [or the epitopes dominant in HLADQ8 + 2 + CD subjects. In 4/6 DQ2 + 8 + subjects there was induction of T-cells specific for homologues of A-gfiadin 57-73. CONCLUSION: Epitopes of gfiadin-specific T-cells are different in HLA-DQ8 and DQ2 associated CD. In CD associated with HLA-DQ2 and DQ8, HLA-DQ2 associated epitopes dominate. Therefore, peptide immunotherapy for CD will be HLA-type specific, or include a cocktail of HLA-DQ2 and DQ8- restricted gliadin epitopes.
128 LowBone Mass in Celiac Disease ls Associated with an Allelic Variation in the ILIB Gene
MariaL. Moreno, Laura Murillo, Horacio Vazquez, Roberto M. Mazure, Alejandra Chemavsky,Alicia Sambuelfi, Edgardo Smecuol, Sonia Niveloni, Silvia Pedreira, M. De Btssio,Eduardo Maurino, Amado S. Pena, Julio C. Bai BACKGROUND/AIM: Celiac disease (CD) is frequently associated with decreased bone mineralmass and an increased rate of fractures in the peripheral skeleton. The interleukin IB(IL-1B)has been implicated in both, the intestinal lesion of CD and the bone remodeling processAn allelic variant of the gene ILl B-511 has been associated with bone demineralizatton in the general population and in patients with Crohn's disease. Our objective was to determinewhether decreased bone mass in CD patients is related with gene polymorphisms encodingfor 1L-lB. MATERIALS & METHODS: We studied 71 CD patients and 60 healthy controlsubjects. At the time of diagnosis, all patients included underwent bone densitometric studies at, either or both, the femoral neck, lumbar spine (L2-L4) and total skeleton. The IL1B-5l1 biallelic polymorphism was genotyped by PCR. RESULTS: (XS• CD patiems have a significantly greater homozygozity for the allele 2 of the [L1B-511 (52%) than that determined in the control population (25%; p<0.01). Compared with non-carriers CD patients,carriers of the allele 2 of the IL1B-511 had significantly lower bone mass (Z-score) atthe total skeleton level (-0.21 • 0.56 vs. - 1.09 - 0.40; p = 0.0488). No statistical differences
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AGA Abstracts