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Langerhans Cells Are Activated In The Esophagus Of Children With Eosinophilic Esophagitis
AB162 Abstracts
J ALLERGY CLIN IMMUNOL FEBRUARY 2010
635
Increasing Prevalence Of Eosinophilic Esophagitis In The Pediatric Population Of Mississippi J. M. Pulcini1, M. Nowicki2, S. Roy1; 1University of Mississippi Medical Center - Department of Pediatric Allergy and Immunology, Jackson, MS, 2 University of Mississippi Medical Center - Department of Pediatric Gastroenterology, Jackson, MS. RATIONALE: Pediatric eosinophilic esophagitis (EoE) has been increasingly diagnosed in the United States. We sought to determine the prevalence of EoE in an area of Southeastern US where the medical literature is lacking. METHODS: We conducted a retrospective review of all esophagogastroduodenoscopies (EGDs) performed by the Division of Pediatric Gastroenterology at the University of Mississippi Medical Center from January 1, 2000 to December 31, 2007. We determined the number of patients diagnosed with reflux esophagitis and eosinophilic esophagitis (EoE) defined as > 20 eosinophils per hpf on esophageal biopsy. RESULTS: Over the 8 year study period, a total of 2,620 EGDs were performed of which 2,013 were diagnostic EGDs. A total of 216 patients were diagnosed with esophagitis by EGD with biopsy during the study period (51 patients with EoE and 165 patients with reflux esophagitis). Characteristics of the EoE patients included: 76% male, 86% Caucasian, age at diagnosis 8.5 years (standard deviation 4 years), and mean 46 eosinophils/hpf. The presenting symptom in EoE patients was gastroesophageal reflux in 71% (36/51) and dysphagia in 24% (12/51). Lastly, the prevalence rate of EoE in patients who underwent diagnostic EGD during the study period was 2.5% and increased significantly during the study period, however the prevalence of GERD was 8.5% and did not increase during the study period. CONCLUSION: In our patient population the prevalence of EoE increased significantly compared to GERD during the study period. Further prospective studies to investigate the racial disparity and increasing prevalence of EOE in our study population are being conducted.
637
636
638
MONDAY
Langerhans Cells Are Activated In The Esophagus Of Children With Eosinophilic Esophagitis M. Chehade, A. Santopolo, M. C. Berin, H. A. Sampson, A. Polydorides, N. Harpaz, O. Yershov, K. Glatman; Mount Sinai School Medicine, New York, NY. RATIONALE: Eosinophilic esophagitis (EoE) is a Th2-mediated inflammatory disease driven by food allergens. The role of esophageal Langerhans cells (LCs) in the pathogenesis of EoE is unknown. Our goal was to determine if LCs are activated in EoE. METHODS: Paraffin-embedded esophageal biopsies from 25 children with EoE and 25 controls were evaluated immunohistochemically. Densities of LCs were determined by expression of CD1a. Co-expression of CD1a and IgE was assessed in a random sample of these subjects (7 EoE, 7 controls). Additionally, expression of IL-4, IL-13, IFN-g, TNF-a, DCLAMP, CCR7, and TSLP in frozen biopsies (9 EoE, 7 controls) was determined by real time PCR. Data were expressed relative to b-actin. RESULTS: IL-4 and IL-13 were significantly upregulated in EoE but undetectable in controls, whereas IFN-g and TNF-a were similar, confirming a Th2 milieu. The LC activation marker DC-LAMP was upregulated in EoE (183634 vs 69616 in EoE vs controls, p<0.05). There was a corresponding decrease in LC density (13 vs 27 LCs/mm2 in EoE vs controls, p50.01), and upregulation of CCR7 (8.362.5 vs 1.460.4 in EoE vs controls, p<0.05). Coexpression of IgE occurred in 19% (range 10-50%) of LCs in EoE epithelium vs 0% in controls. All EoE biopsies contained IgE-positive LCs. TSLP, which induces a Th2-skewing phenotype in LCs, was upregulated in EoE (15926390 vs 305633 in EoE vs controls, p<0.01). CONCLUSIONS: Our results demonstrate phenotypic evidence of activation and migration in esophageal LCs of pediatric patients with EoE. Triggers may include cross-linking of IgE via the high-affinity receptor and stimulation by TSLP.
Glucocorticoid-mediated FKBP51 Gene Expression in Esophageal Epithelial Cells J. M. Caldwell, C. Blanchard, M. E. Rothenberg; Cincinnati Children’s Hospital Medical Center, Cincinnati, OH. RATIONALE: We aimed to determine the molecular mechanism by which glucocorticoid treatment controls the expression of FK506-binding protein 51 (FKBP51) in esophageal epithelial cells. METHODS: We identified glucocorticoid-regulated transcripts in the esophagus of eosinophilic esophagitis patients who were treated with swallowed fluticasone propionate. FKBP51 mRNA and protein expression patterns were monitored in esophageal epithelial cells treated with glucocorticoid receptor (GR) agonists, GR antagonist, and inhibitors of de novo protein synthesis and transcription. RESULTS: FK506-binding protein 51 was identified as a gene upregulated in the esophagus of EE patients who were treated with and responded to swallowed fluticasone propionate (FP). FKBP51 has been shown to be a glucocorticoid-induced gene in lymphocytes and respiratory epithelial cells, but its role in the esophagus has not been studied. We observed that glucocorticoid treatment of primary esophageal epithelial cells and the esophageal cell line TE-7 increased FKBP51 mRNA and protein expression. We found that this depended on glucocorticoid receptor signaling as co-treatment of cells with the glucocorticoid receptor antagonist RU486 abolished the increased FKBP51 expression. Furthermore, the glucocorticoid-mediated increase in FKBP51 mRNA did not require de novo protein synthesis. Increased transcript stability did not account for the increased FKBP51 expression, as the half-life of FKBP51 mRNA was 8-9 h in the presence or absence of glucocorticoid treatment. CONCLUSIONS: These data support the interpretation that glucocorticoid receptor signaling directly mediates increased FKBP51 expression in esophageal epithelial cells. Because glucocorticoid treatment does not impact the half-life of FKBP51 mRNA, glucocorticoid receptor signaling likely increases FKBP51 promoter activity in esophageal epithelial cells.
Fibroblast Growth Factor as an Improved Disease Indicator for Eosinophilic Esophagitis Compared with Eotaxin-3 and IL-5 J. J. Huang, T. Nguyen, K. C. Nadeau; Stanford University, Stanford, CA. RATIONALE: Eosinophilic esophagitis (EoE) is associated with dysphagia and possible strictures in the esophagus. As part of the diagnostic work up, gastroesophageal reflux disease (GERD) is ruled out as a primary disease. EoE patients are diagnosed through esophageal biopsy with counts of 15 or more eosinophils per high-powered field. Although eotaxin-3 has been correlated with EoE, more specific and sensitive plasma predictive factors for EoE are needed. METHODS: Cytometric bead array (BD Biosciences, San Jose, CA) was performed in duplicate to study plasma cytokines of subjects with EoE (n512), GERD (n58), eosinophilic colitis (EOC) (n53), eosinophilic gastritis (EOG) (n53), inflammatory bowel disease (IBD) (ulcerative colitis, n53 and Crohn’s Disease, n53), and healthy controls (HC) (n58). Cytokines analyzed included eotaxin-3, fibroblast growth factor basic (FGFb), G-CSF, IFN-g, IL-17, IL-1a, IL4, IL-5, macrophage inflammatory protein 1a, and nerve growth factor. Statistics were performed (Mann-Whitney U test). RESULTS: FGFb is significantly up-regulated in EoE compared to GERD, EOC/EOG, IBD, and HC (p<0.05). FGFb had a positive predictive value of 1 compared with eotaxin-3, which had a positive predictive value of 0.8 when differentiating EoE from GERD, IBD, and HC patients. FGFb was less elevated in EOC/EOG compared to EoE patients. IL-5 was only found to be significantly increased (p<0.05) in EoE vs. GERD but not vs. EOC/EOG, IBD, or HC. CONCLUSIONS: FGFb may be used as a predictive factor for EoE. More research on FGFb as a disease indicator is needed. FGFb could be associated with the pathophysiological mechanism of EoE.
J ALLERGY CLIN IMMUNOL FEBRUARY 2010
635
Increasing Prevalence Of Eosinophilic Esophagitis In The Pediatric Population Of Mississippi J. M. Pulcini1, M. Nowicki2, S. Roy1; 1University of Mississippi Medical Center - Department of Pediatric Allergy and Immunology, Jackson, MS, 2 University of Mississippi Medical Center - Department of Pediatric Gastroenterology, Jackson, MS. RATIONALE: Pediatric eosinophilic esophagitis (EoE) has been increasingly diagnosed in the United States. We sought to determine the prevalence of EoE in an area of Southeastern US where the medical literature is lacking. METHODS: We conducted a retrospective review of all esophagogastroduodenoscopies (EGDs) performed by the Division of Pediatric Gastroenterology at the University of Mississippi Medical Center from January 1, 2000 to December 31, 2007. We determined the number of patients diagnosed with reflux esophagitis and eosinophilic esophagitis (EoE) defined as > 20 eosinophils per hpf on esophageal biopsy. RESULTS: Over the 8 year study period, a total of 2,620 EGDs were performed of which 2,013 were diagnostic EGDs. A total of 216 patients were diagnosed with esophagitis by EGD with biopsy during the study period (51 patients with EoE and 165 patients with reflux esophagitis). Characteristics of the EoE patients included: 76% male, 86% Caucasian, age at diagnosis 8.5 years (standard deviation 4 years), and mean 46 eosinophils/hpf. The presenting symptom in EoE patients was gastroesophageal reflux in 71% (36/51) and dysphagia in 24% (12/51). Lastly, the prevalence rate of EoE in patients who underwent diagnostic EGD during the study period was 2.5% and increased significantly during the study period, however the prevalence of GERD was 8.5% and did not increase during the study period. CONCLUSION: In our patient population the prevalence of EoE increased significantly compared to GERD during the study period. Further prospective studies to investigate the racial disparity and increasing prevalence of EOE in our study population are being conducted.
637
636
638
MONDAY
Langerhans Cells Are Activated In The Esophagus Of Children With Eosinophilic Esophagitis M. Chehade, A. Santopolo, M. C. Berin, H. A. Sampson, A. Polydorides, N. Harpaz, O. Yershov, K. Glatman; Mount Sinai School Medicine, New York, NY. RATIONALE: Eosinophilic esophagitis (EoE) is a Th2-mediated inflammatory disease driven by food allergens. The role of esophageal Langerhans cells (LCs) in the pathogenesis of EoE is unknown. Our goal was to determine if LCs are activated in EoE. METHODS: Paraffin-embedded esophageal biopsies from 25 children with EoE and 25 controls were evaluated immunohistochemically. Densities of LCs were determined by expression of CD1a. Co-expression of CD1a and IgE was assessed in a random sample of these subjects (7 EoE, 7 controls). Additionally, expression of IL-4, IL-13, IFN-g, TNF-a, DCLAMP, CCR7, and TSLP in frozen biopsies (9 EoE, 7 controls) was determined by real time PCR. Data were expressed relative to b-actin. RESULTS: IL-4 and IL-13 were significantly upregulated in EoE but undetectable in controls, whereas IFN-g and TNF-a were similar, confirming a Th2 milieu. The LC activation marker DC-LAMP was upregulated in EoE (183634 vs 69616 in EoE vs controls, p<0.05). There was a corresponding decrease in LC density (13 vs 27 LCs/mm2 in EoE vs controls, p50.01), and upregulation of CCR7 (8.362.5 vs 1.460.4 in EoE vs controls, p<0.05). Coexpression of IgE occurred in 19% (range 10-50%) of LCs in EoE epithelium vs 0% in controls. All EoE biopsies contained IgE-positive LCs. TSLP, which induces a Th2-skewing phenotype in LCs, was upregulated in EoE (15926390 vs 305633 in EoE vs controls, p<0.01). CONCLUSIONS: Our results demonstrate phenotypic evidence of activation and migration in esophageal LCs of pediatric patients with EoE. Triggers may include cross-linking of IgE via the high-affinity receptor and stimulation by TSLP.
Glucocorticoid-mediated FKBP51 Gene Expression in Esophageal Epithelial Cells J. M. Caldwell, C. Blanchard, M. E. Rothenberg; Cincinnati Children’s Hospital Medical Center, Cincinnati, OH. RATIONALE: We aimed to determine the molecular mechanism by which glucocorticoid treatment controls the expression of FK506-binding protein 51 (FKBP51) in esophageal epithelial cells. METHODS: We identified glucocorticoid-regulated transcripts in the esophagus of eosinophilic esophagitis patients who were treated with swallowed fluticasone propionate. FKBP51 mRNA and protein expression patterns were monitored in esophageal epithelial cells treated with glucocorticoid receptor (GR) agonists, GR antagonist, and inhibitors of de novo protein synthesis and transcription. RESULTS: FK506-binding protein 51 was identified as a gene upregulated in the esophagus of EE patients who were treated with and responded to swallowed fluticasone propionate (FP). FKBP51 has been shown to be a glucocorticoid-induced gene in lymphocytes and respiratory epithelial cells, but its role in the esophagus has not been studied. We observed that glucocorticoid treatment of primary esophageal epithelial cells and the esophageal cell line TE-7 increased FKBP51 mRNA and protein expression. We found that this depended on glucocorticoid receptor signaling as co-treatment of cells with the glucocorticoid receptor antagonist RU486 abolished the increased FKBP51 expression. Furthermore, the glucocorticoid-mediated increase in FKBP51 mRNA did not require de novo protein synthesis. Increased transcript stability did not account for the increased FKBP51 expression, as the half-life of FKBP51 mRNA was 8-9 h in the presence or absence of glucocorticoid treatment. CONCLUSIONS: These data support the interpretation that glucocorticoid receptor signaling directly mediates increased FKBP51 expression in esophageal epithelial cells. Because glucocorticoid treatment does not impact the half-life of FKBP51 mRNA, glucocorticoid receptor signaling likely increases FKBP51 promoter activity in esophageal epithelial cells.
Fibroblast Growth Factor as an Improved Disease Indicator for Eosinophilic Esophagitis Compared with Eotaxin-3 and IL-5 J. J. Huang, T. Nguyen, K. C. Nadeau; Stanford University, Stanford, CA. RATIONALE: Eosinophilic esophagitis (EoE) is associated with dysphagia and possible strictures in the esophagus. As part of the diagnostic work up, gastroesophageal reflux disease (GERD) is ruled out as a primary disease. EoE patients are diagnosed through esophageal biopsy with counts of 15 or more eosinophils per high-powered field. Although eotaxin-3 has been correlated with EoE, more specific and sensitive plasma predictive factors for EoE are needed. METHODS: Cytometric bead array (BD Biosciences, San Jose, CA) was performed in duplicate to study plasma cytokines of subjects with EoE (n512), GERD (n58), eosinophilic colitis (EOC) (n53), eosinophilic gastritis (EOG) (n53), inflammatory bowel disease (IBD) (ulcerative colitis, n53 and Crohn’s Disease, n53), and healthy controls (HC) (n58). Cytokines analyzed included eotaxin-3, fibroblast growth factor basic (FGFb), G-CSF, IFN-g, IL-17, IL-1a, IL4, IL-5, macrophage inflammatory protein 1a, and nerve growth factor. Statistics were performed (Mann-Whitney U test). RESULTS: FGFb is significantly up-regulated in EoE compared to GERD, EOC/EOG, IBD, and HC (p<0.05). FGFb had a positive predictive value of 1 compared with eotaxin-3, which had a positive predictive value of 0.8 when differentiating EoE from GERD, IBD, and HC patients. FGFb was less elevated in EOC/EOG compared to EoE patients. IL-5 was only found to be significantly increased (p<0.05) in EoE vs. GERD but not vs. EOC/EOG, IBD, or HC. CONCLUSIONS: FGFb may be used as a predictive factor for EoE. More research on FGFb as a disease indicator is needed. FGFb could be associated with the pathophysiological mechanism of EoE.