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Retrospective Chart Review to Correlate Airborne Allergen Skin Test Results with Biopsy Documented Esophageal Eosinophil Count in Eosinophilic Esophagitis (EOE)
Abstracts AB161
J ALLERGY CLIN IMMUNOL VOLUME 125, NUMBER 2
A Model of Eosinophilic Esophagitis (EE) and Villus Atrophy (VA) after Challenge in Mice Sensitized to Peanuts: Improvement by Epicutaneous Immunotherapy (EPIT) L. Mondoulet1, V. Dioszeghy1, M. Ligouis1, V. Dhelft1, T. Larcher2, Y. Cherel2, C. Dupont3, P. Benhamou1; 1DBV Technologies, PARIS, FRANCE, 2APEX, INRA, National Veterinary School, Nantes, FRANCE, 3 Paris-Descartes, Saint Vincent de Paul Hospital, PARIS, FRANCE. RATIONALE: Food allergy may trigger EE and VA in humans. EPIT has been described as a therapeutic method in food allergy. We developed a model of EE and VA sensitized mice exclusively fed by peanut protein extracts (PPE) using to evaluate EPITas a preventive treatment of EE and VA. METHODS: After oral sensitization with PPE and cholera toxin, 30 BALB/c mice were treated weekly during 8 weeks by PPE skin applications (EPIT), 20 mice were not treated (NT) and 10 mice constituted the control group (C). Mice were then exclusively fed with PPE. Specific IgE, IgG1 and IgG2a were monitored every 2 weeks. Esophageal and jejunal samples were taken for histology. RESULTS: sIgE increased after oral sensitization, respectively 0.20760.03 and 0.21460.04 mg/ml, in EPIT and NT, with undetectable values in C. Following EPIT, sIgE decreased and sIgG2a increased, respectively 0.13960.01 vs 0.16660.01 mg/ml (EPIT vs NT, p<0.05) and 14.9660.60 vs 4.7361.75 mg/ml (p<0.05). Esophageal eosinophilic infiltration (measured in 6 high power fields) was higher in NT, 136632, than in EPIT, 50612 (p<0.05) and C, 763 cells/mm2 (p<0.01). Esophagus mucosa thickness was increased in NT compared to EPIT (p<0.01) and NT (p<0.001). The duodenal villus/crypt ratio was lower in NT than in EPIT and C, respectively 1.660.1 vs 2.360.2 (p<0.01) and 2.460.1 (p<0.001). Eosinophilic infiltration in jejunum was increased in NT compared to EPIT (p<0.01) and C (p<0.001). CONCLUSIONS: EPIT seems efficient to prevent EE and VA induced by PPE exclusive feeding in previously sensitized mice.
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Epithelial Mesenchymal Transition in Eosinophilic Esophagitis: Identification and Contributions to Esophageal Remodeling and Fibrosis N. Akhtar1, S. A. Woodruff2, V. Mukkada2, S. Fillon2, J. Masterson2, A. F. Kagalwalla1,3, C. Jun1, J. J. Lee4, G. T. Furuta2, S. J. Ackerman1; 1 University of Illinois at Chicago, Chicago, IL, 2University of Colorado, Denver Children’s Hospital, Denver, CO, 3Children’s Memorial Hospital, Northwestern University, Chicago, IL, 4Mayo Clinic, Scottsdale, AZ. RATIONALE: Esophageal remodeling in Eosinophilic Esophagitis (EoE), a food-allergic disease, includes epithelial hyperplasia and subepithelial fibrosis. Epithelial Mesenchymal Transition (EMT) is a process whereby non-motile epithelial cells de-differentiate into motile mesenchymal-like cells during chronic inflammation, and can further differentiate into myofibroblasts. Since the mechanism of esophageal fibrosis in EoE is poorly understood, our objective was to determine whether EMT occurs in EoE and contributes to its pathogenesis. METHODS: Sections from 60 formalin-fixed, paraffin-embedded biopsies from patients with EoE (17), GERD (7), indeterminate esophagitis (15), and normal esophagus (21) were evaluated for EMT by two-color immunofluorescence staining for cells expressing cytokeratin (epithelial) and vimentin (mesenchymal) markers. Sections were analyzed in a blinded fashion by confocal microscopy and graded for the presence/degree of EMT using a 6-point scale. Mean EMT indices/hpf were analyzed for relationships to diagnosis, eosinophils/hpf, and indices for eosinophil peroxidase (EPX) and TGF-beta immunostaining, and esophageal fibrosis. RESULTS: The highest EMT index was associated with a patient diagnosis of EoE (3.2560.28), followed by indeterminate esophagitis(2.6960.29)>GERD(1.6260.39)>normal(1.0660.15). EMT was positively correlated (all p<0.01) with eosinophils/hpf (r50.691), EPX (r50.738) and TGF-beta (r50.520) staining indices, and esophageal fibrosis scores (r50.644). CONCLUSIONS: This is the first identification of EMT in EoE. EMT was observed principally in EoE, to a lesser degree in indeterminate esophagitis with fibrosis, whereas it was observed only once in GERD, and not in
normal esophagus. The significant correlations of EMT to esophageal eosinophils and their activation, and measures of fibrosis, suggest that EMT may contribute to the subepithelial fibrosis characteristic of EoE pathogenesis.
633
Clinical Course of Eosinophilic Gastrointestinal Diseases (EGID) with and without Eosinophilia: A Retrospective Case Review J. Lee, C. R. Weiler, J. A. Alexander; Mayo Clinic Rochester, Rochester, MN. RATIONALE: Hypereosinophilic syndrome (HES) is diagnosed by excluding secondary causes of persistent significant eosinophilia (>1500 eos/mL) with end organ damage/dysfunction. The clinical course of eosinophilic gastrointestinal diseases (EGID) with significant eosinophilia, considered an HES ‘‘overlap variant,’’ is poorly defined. METHODS: We performed a retrospective chart review of 41 individuals with biopsy-proven EGID, identified from Mayo Clinic Rochester electronic records (1995 to 2008), with peripheral eosinophil count above (group A) and below (group B) 1500 eos/mL. Patients with only esophageal disease were excluded. RESULTS: All presented to gastroenterology and most were followed by allergy (A: 71%; B: 63%). Group A (n514) had a 3-fold higher peripheral eosinophilia (2834 vs 856 eos/mL; normal<500/mL) and higher frequency of > 1 diseased GI segment (esophagus, stomach, small bowel) than group B (n527) (p50.0071). There was no evidence of eosinophilic end organ damage on available bone marrow biopsies and echocardiograms (A: 10 and 7; B: 7 and 6). Prednisone response was more common in group A (12/12 vs. 17/22). Median doses of initial (40 mg/day for both) and maintenance prednisone (10 mg and 5 mg) were similar, with a tendency for longer treatment duration (1.75 months vs. 1 month; 14 months vs. 9 months) for group A. Time to recurrence ranged widely from days to 10 months in both groups. Non-steroidal agents were more frequently employed in group A. CONCLUSIONS: EGID with peripheral eosinophilia is associated with greater GI segmental involvement and may be more responsive to prednisone. However, recurrence was common, underscoring the need for targeted therapies.
634
Retrospective Chart Review to Correlate Airborne Allergen Skin Test Results with Biopsy Documented Esophageal Eosinophil Count in Eosinophilic Esophagitis (EOE) C. W. Bassett1,2, E. Rothstein1, Y. Kopyltsova1, A. Stern1; 1NYU School of Medicine, New York, NY, 2Allergy and ASthma Care of NY, New York, NY. RATIONALE: Studies suggest that airborne allergens have been implicated as possible inciting agents in addition to foods in eosinophilic esophagitis (EE). A possible mechanism may include initial allergen sensitization in the respiratory tract as well as in the esophagus. METHODS: Retrospective case series of 9 patients with biopsy proven eosinophilic esophagitis were reviewed for their response to food and airborne allergens. Degree of airborne sensitivity was then correlated with biopsy documented esophageal eosinophil count. RESULTS: Diagnostic airborne allergy skin tests were positive and significant in 88% of EoE. Additionally, of the patients reviewed, 8 patients had >5 15 eosinophils/HPF documented through biopsy. Of these 8 patients, 5 had a moderate or higher response to airborne allergens (63%), and 3 had a mild response to airborne allergens (37%). Of the 3 patients who had >5 40 eosinophils/HPF, all (100%) had a moderate or severe response to airborne allergens. CONCLUSIONS: Patients with history of atopy, may be at an increased risk for developing eosinophilic esophagitis. Higher degrees of airborne sensitization also correlate with greater eosinophilic counts on esophageal biopsy - as those patients with the highest eosinophilic counts (>5 40 eosinophils/HPF) had the greatest sensitivity to airborne allergies in this retrospective review of patients with EoE.
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J ALLERGY CLIN IMMUNOL VOLUME 125, NUMBER 2
A Model of Eosinophilic Esophagitis (EE) and Villus Atrophy (VA) after Challenge in Mice Sensitized to Peanuts: Improvement by Epicutaneous Immunotherapy (EPIT) L. Mondoulet1, V. Dioszeghy1, M. Ligouis1, V. Dhelft1, T. Larcher2, Y. Cherel2, C. Dupont3, P. Benhamou1; 1DBV Technologies, PARIS, FRANCE, 2APEX, INRA, National Veterinary School, Nantes, FRANCE, 3 Paris-Descartes, Saint Vincent de Paul Hospital, PARIS, FRANCE. RATIONALE: Food allergy may trigger EE and VA in humans. EPIT has been described as a therapeutic method in food allergy. We developed a model of EE and VA sensitized mice exclusively fed by peanut protein extracts (PPE) using to evaluate EPITas a preventive treatment of EE and VA. METHODS: After oral sensitization with PPE and cholera toxin, 30 BALB/c mice were treated weekly during 8 weeks by PPE skin applications (EPIT), 20 mice were not treated (NT) and 10 mice constituted the control group (C). Mice were then exclusively fed with PPE. Specific IgE, IgG1 and IgG2a were monitored every 2 weeks. Esophageal and jejunal samples were taken for histology. RESULTS: sIgE increased after oral sensitization, respectively 0.20760.03 and 0.21460.04 mg/ml, in EPIT and NT, with undetectable values in C. Following EPIT, sIgE decreased and sIgG2a increased, respectively 0.13960.01 vs 0.16660.01 mg/ml (EPIT vs NT, p<0.05) and 14.9660.60 vs 4.7361.75 mg/ml (p<0.05). Esophageal eosinophilic infiltration (measured in 6 high power fields) was higher in NT, 136632, than in EPIT, 50612 (p<0.05) and C, 763 cells/mm2 (p<0.01). Esophagus mucosa thickness was increased in NT compared to EPIT (p<0.01) and NT (p<0.001). The duodenal villus/crypt ratio was lower in NT than in EPIT and C, respectively 1.660.1 vs 2.360.2 (p<0.01) and 2.460.1 (p<0.001). Eosinophilic infiltration in jejunum was increased in NT compared to EPIT (p<0.01) and C (p<0.001). CONCLUSIONS: EPIT seems efficient to prevent EE and VA induced by PPE exclusive feeding in previously sensitized mice.
632
Epithelial Mesenchymal Transition in Eosinophilic Esophagitis: Identification and Contributions to Esophageal Remodeling and Fibrosis N. Akhtar1, S. A. Woodruff2, V. Mukkada2, S. Fillon2, J. Masterson2, A. F. Kagalwalla1,3, C. Jun1, J. J. Lee4, G. T. Furuta2, S. J. Ackerman1; 1 University of Illinois at Chicago, Chicago, IL, 2University of Colorado, Denver Children’s Hospital, Denver, CO, 3Children’s Memorial Hospital, Northwestern University, Chicago, IL, 4Mayo Clinic, Scottsdale, AZ. RATIONALE: Esophageal remodeling in Eosinophilic Esophagitis (EoE), a food-allergic disease, includes epithelial hyperplasia and subepithelial fibrosis. Epithelial Mesenchymal Transition (EMT) is a process whereby non-motile epithelial cells de-differentiate into motile mesenchymal-like cells during chronic inflammation, and can further differentiate into myofibroblasts. Since the mechanism of esophageal fibrosis in EoE is poorly understood, our objective was to determine whether EMT occurs in EoE and contributes to its pathogenesis. METHODS: Sections from 60 formalin-fixed, paraffin-embedded biopsies from patients with EoE (17), GERD (7), indeterminate esophagitis (15), and normal esophagus (21) were evaluated for EMT by two-color immunofluorescence staining for cells expressing cytokeratin (epithelial) and vimentin (mesenchymal) markers. Sections were analyzed in a blinded fashion by confocal microscopy and graded for the presence/degree of EMT using a 6-point scale. Mean EMT indices/hpf were analyzed for relationships to diagnosis, eosinophils/hpf, and indices for eosinophil peroxidase (EPX) and TGF-beta immunostaining, and esophageal fibrosis. RESULTS: The highest EMT index was associated with a patient diagnosis of EoE (3.2560.28), followed by indeterminate esophagitis(2.6960.29)>GERD(1.6260.39)>normal(1.0660.15). EMT was positively correlated (all p<0.01) with eosinophils/hpf (r50.691), EPX (r50.738) and TGF-beta (r50.520) staining indices, and esophageal fibrosis scores (r50.644). CONCLUSIONS: This is the first identification of EMT in EoE. EMT was observed principally in EoE, to a lesser degree in indeterminate esophagitis with fibrosis, whereas it was observed only once in GERD, and not in
normal esophagus. The significant correlations of EMT to esophageal eosinophils and their activation, and measures of fibrosis, suggest that EMT may contribute to the subepithelial fibrosis characteristic of EoE pathogenesis.
633
Clinical Course of Eosinophilic Gastrointestinal Diseases (EGID) with and without Eosinophilia: A Retrospective Case Review J. Lee, C. R. Weiler, J. A. Alexander; Mayo Clinic Rochester, Rochester, MN. RATIONALE: Hypereosinophilic syndrome (HES) is diagnosed by excluding secondary causes of persistent significant eosinophilia (>1500 eos/mL) with end organ damage/dysfunction. The clinical course of eosinophilic gastrointestinal diseases (EGID) with significant eosinophilia, considered an HES ‘‘overlap variant,’’ is poorly defined. METHODS: We performed a retrospective chart review of 41 individuals with biopsy-proven EGID, identified from Mayo Clinic Rochester electronic records (1995 to 2008), with peripheral eosinophil count above (group A) and below (group B) 1500 eos/mL. Patients with only esophageal disease were excluded. RESULTS: All presented to gastroenterology and most were followed by allergy (A: 71%; B: 63%). Group A (n514) had a 3-fold higher peripheral eosinophilia (2834 vs 856 eos/mL; normal<500/mL) and higher frequency of > 1 diseased GI segment (esophagus, stomach, small bowel) than group B (n527) (p50.0071). There was no evidence of eosinophilic end organ damage on available bone marrow biopsies and echocardiograms (A: 10 and 7; B: 7 and 6). Prednisone response was more common in group A (12/12 vs. 17/22). Median doses of initial (40 mg/day for both) and maintenance prednisone (10 mg and 5 mg) were similar, with a tendency for longer treatment duration (1.75 months vs. 1 month; 14 months vs. 9 months) for group A. Time to recurrence ranged widely from days to 10 months in both groups. Non-steroidal agents were more frequently employed in group A. CONCLUSIONS: EGID with peripheral eosinophilia is associated with greater GI segmental involvement and may be more responsive to prednisone. However, recurrence was common, underscoring the need for targeted therapies.
634
Retrospective Chart Review to Correlate Airborne Allergen Skin Test Results with Biopsy Documented Esophageal Eosinophil Count in Eosinophilic Esophagitis (EOE) C. W. Bassett1,2, E. Rothstein1, Y. Kopyltsova1, A. Stern1; 1NYU School of Medicine, New York, NY, 2Allergy and ASthma Care of NY, New York, NY. RATIONALE: Studies suggest that airborne allergens have been implicated as possible inciting agents in addition to foods in eosinophilic esophagitis (EE). A possible mechanism may include initial allergen sensitization in the respiratory tract as well as in the esophagus. METHODS: Retrospective case series of 9 patients with biopsy proven eosinophilic esophagitis were reviewed for their response to food and airborne allergens. Degree of airborne sensitivity was then correlated with biopsy documented esophageal eosinophil count. RESULTS: Diagnostic airborne allergy skin tests were positive and significant in 88% of EoE. Additionally, of the patients reviewed, 8 patients had >5 15 eosinophils/HPF documented through biopsy. Of these 8 patients, 5 had a moderate or higher response to airborne allergens (63%), and 3 had a mild response to airborne allergens (37%). Of the 3 patients who had >5 40 eosinophils/HPF, all (100%) had a moderate or severe response to airborne allergens. CONCLUSIONS: Patients with history of atopy, may be at an increased risk for developing eosinophilic esophagitis. Higher degrees of airborne sensitization also correlate with greater eosinophilic counts on esophageal biopsy - as those patients with the highest eosinophilic counts (>5 40 eosinophils/HPF) had the greatest sensitivity to airborne allergies in this retrospective review of patients with EoE.
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