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Our 3-year Experience with Patch Testing for Eosinophilic Esophagitis
Abstracts AB93
J ALLERGY CLIN IMMUNOL VOLUME 129, NUMBER 2
Identification Of Increased Oral Eosinophils In Patients With Eosinophilic Esophagitis (EE) Using Oral Rinse Analysis: Proof Of Concept J. Ohayon; McMaster University, Hamilton, ON, CANADA. RATIONALE: Patients with suspected EE require endoscopic biopsy for confirmation. Evaluations of oral rinses from EE patients were assessed for increased Eosinophils (Eos) as a possible screening tool. METHODS: EE Patients were asked to rinse with 10 mls of saline for 30 seconds. Expectorates were fixed with formaldehyde. Following filtration to remove epithelial cells, samples were centrifuged at 3000 rpm. Cell pellets were re-suspended in PBS. 50 microliters of cells were stained with Hematoxylin and Eosin. Eos and neutrophils (PMN) were visualized and quantified under light microscopy by characteristic nuclei, eosin containing granules and size. Quantification of Eos was assessed in microscopic fields. Control samples were obtained. RESULTS: Six patients with biopsy proven EE showed an average of 19% Eos (range 10-12%) of all cells evaluated. This was significantly more than control, (mean 3.1% range 2-4%). There was a higher absolute number of PMN’s in control patients as compared to patients with EE (111 vs. 49). CONCLUSIONS: There is evidence of increased amount of Eos isolated from oral rinses of EE patients. A combination of increased Eos and decreased PMN’s resulted in an increase relative proportion of Eos in patients with EE. The analysis of oral rinses may serve as a screening tool for patients suspected of having EE.
353
Fecal Eosinophil-derived Neurotoxin Is Significantly Elevated In Non-ige Dependent Gastrointestinal Allergies, Especially In Subtypes Showing Bloody Stool H. Morita1,2, I. Nomura1, T. Shoda1, H. Saito1, K. Matsumoto1; 1National Research Institute for Child Health and Development, Tokyo, JAPAN, 2 Department of Pediatrics, Keio University School of Medicine, Tokyo, JAPAN. RATIONALE: Gastrointestinal allergies (GI-allergies), including food protein-induced enterocolitis syndrome (FPIES), proctocolitis and enteropathy are thought to be non-IgE-associated and cell-mediated food allergies. However, the precise pathogenesis of these allergies remains uncertain. In order to determine whether eosinophils are involved in these GI-allergies or not, we measured eosinophil-derived neurotoxin (EDN) levels in stool samples from patients with various GI-allergies. METHODS: Stool samples were obtained from patients with GI-allergies (n542) or healthy infants (n5230). Stool sample extracts were prepared using sample preparation kits (Roche) and the EDN levels in the stool sample extracts were measured by ELISA. RESULTS: The EDN levels in the stool sample extracts from patients with GI-allergies were significantly higher than those from healthy infant. In addition, the EDN levels in the stool sample extracts varied among GIallergies; the EDN levels in the stool sample extract from patients who exhibited bloody stools were higher than those without bloody stools. CONCLUSIONS: Our results suggest that eosinophils are likely to be involved in the pathogenesis of GI-allergies, in particular, those with bloody stools. Our results also imply the involvement of Th2-type immune responses in the pathogenesis of GI-allergies through activation of eosinophils but not IgE synthesis.
354
Dynamics of Eosinophils in Non-IgE-mediated Gastrointestinal Food Allergies in Neonates and Infants, differences between 4 Clusters I. Nomura, H. Morita, T. Shoda, K. Arai, N. Ito, A. Nakazawa, Y. Ohya, H. Saito, K. Matsumoto; National Center for Child Health and Development, Tokyo, JAPAN. RATIONALE: Non-IgE-mediated gastrointestinal food allergies are consisted from several syndromes; FPIES, proctocolitis, enteropathy and allergic eosinophilic gastroenteritis (AEG). AEG and enteropathy can be diagnosed only by histological examination. To establish effective initial diagnosis of whole patients at first presentation, we tried to classify those by simple clinical variables and then, 4 clusters were derived (JACI 2011:127:685-688). In this paper, we tried to make comparison of eosinophils, important biological marker in GI allergy, between those 4 clusters. METHODS: Patients (total 300) were registered to database of our research group from 2007 to 2011. Clusters were defined according to presence of vomiting and/or bloody stool in the first 0-2 months after onset; Cluster 1 (vomiting +/ bloody stool +), Cluster 2 (+/-), Cluster 3 (-/-), Cluster 4 (-/+). Gastrointestinal histology; thirteen patients were examined their entire GI tract by fiberscope. Fecal eosinophils; mucous part of the feces was taken and stained with Hansel’s staining kit. RESULTS: Circulating eosinophil counts; highest value was seen in Cluster 3 (p<0.01), 14% of whole patients showed more than 30% of circulating eosinophils. Gastrointestinal histology; in Cluster 3 and 4, marked infiltration of eosinophils were admitted. In Cluster 3, inflammation was spread over entire GI tract but epithelial damage was minimal. Fecal eosinophils; high in Cluster 1 and 4, while Cluster 3 showed within normal limit. Data was confirmed by determination of fecal eosinophilderived neurotoxin. CONCLUSIONS: Difference in dynamics of eosinophils among 4 clusters was clarified. It might be very beneficial to conduct diagnosis and treatment considering this dynamics.
355
Our 3-year Experience with Patch Testing for Eosinophilic Esophagitis C. S. Bauer1, A. R. Lamba1, H. P. Zimmerman1, R. G. Hoffmann1, M. Dasgupta1, R. J. Noel1, M. B. Levy2, M. Vasudev1; 1Medical College of Wisconsin, Milwaukee, WI, 2Assaf Harofe Medical Center, Tzrifin, ISRAEL. RATIONALE: Treatment of eosinophilic esophagitis (EoE) consists of pharmacotherapy and/or dietary therapy. Food atopy patch testing (APT) for EoE is not standardized and its clinical utility is controversial. We hypothesize that our APT method is valuable adjunctive therapy to aid in treatment of pediatric EoE. METHODS: Sixty-two patients (male549, mean age58.8 years) with eosinophils on esophageal biopsy (mean proximally538/HPF, distally541/HPF) who were evaluated by food APT between 01/2008-02/ 2011 were retrospectively identified by ICD-9 codes. APT was performed using powdered food, sterile water, and glycerin in a 12-food EoE panel. Biopsies, food-allergy tests [serum IgE (sIgE), percutaneous skin test (PST), APT], and swallowed steroid (SS) usage were recorded. _1 food in 74%. For APT, RESULTS: Combined APT and PST identified > 53% (n533/62) were positive [peanut (n513/55, 24%), corn (n514/61, 23%), and chicken (n511/58, 19%)], for PST, 57% (n535/61; most commonly, peanut, wheat, and corn), and for sIgE, 67% (n512/18). Eleven patients had negative PST, but positive APT. Those with surveillance biopsies (n539) had 2 on average; 39% of surveillance biopsies (n531/79) were on guided-elimination (GE) from food-allergy testing. Those on SS (n512/39, 30.8%), SS and GE (n512/39, 30.8%), and GE (10.2%, n54/39) had 62% (proximally) and 69% (distally) decrease in mean eosinophils/HPF on first surveillance biopsy after APT compared to diagnostic biopsy. 56% (n510/18) of those on APT-based GE had <15/ eosinophils/HPF on first surveillance biopsy, though 70% of those (n57/ 10) were concomitantly on PST-based GE and SS. CONCLUSIONS: Our APT method with subsequent food elimination aids in the complex treatment of pediatric EoE.
SUNDAY
352
J ALLERGY CLIN IMMUNOL VOLUME 129, NUMBER 2
Identification Of Increased Oral Eosinophils In Patients With Eosinophilic Esophagitis (EE) Using Oral Rinse Analysis: Proof Of Concept J. Ohayon; McMaster University, Hamilton, ON, CANADA. RATIONALE: Patients with suspected EE require endoscopic biopsy for confirmation. Evaluations of oral rinses from EE patients were assessed for increased Eosinophils (Eos) as a possible screening tool. METHODS: EE Patients were asked to rinse with 10 mls of saline for 30 seconds. Expectorates were fixed with formaldehyde. Following filtration to remove epithelial cells, samples were centrifuged at 3000 rpm. Cell pellets were re-suspended in PBS. 50 microliters of cells were stained with Hematoxylin and Eosin. Eos and neutrophils (PMN) were visualized and quantified under light microscopy by characteristic nuclei, eosin containing granules and size. Quantification of Eos was assessed in microscopic fields. Control samples were obtained. RESULTS: Six patients with biopsy proven EE showed an average of 19% Eos (range 10-12%) of all cells evaluated. This was significantly more than control, (mean 3.1% range 2-4%). There was a higher absolute number of PMN’s in control patients as compared to patients with EE (111 vs. 49). CONCLUSIONS: There is evidence of increased amount of Eos isolated from oral rinses of EE patients. A combination of increased Eos and decreased PMN’s resulted in an increase relative proportion of Eos in patients with EE. The analysis of oral rinses may serve as a screening tool for patients suspected of having EE.
353
Fecal Eosinophil-derived Neurotoxin Is Significantly Elevated In Non-ige Dependent Gastrointestinal Allergies, Especially In Subtypes Showing Bloody Stool H. Morita1,2, I. Nomura1, T. Shoda1, H. Saito1, K. Matsumoto1; 1National Research Institute for Child Health and Development, Tokyo, JAPAN, 2 Department of Pediatrics, Keio University School of Medicine, Tokyo, JAPAN. RATIONALE: Gastrointestinal allergies (GI-allergies), including food protein-induced enterocolitis syndrome (FPIES), proctocolitis and enteropathy are thought to be non-IgE-associated and cell-mediated food allergies. However, the precise pathogenesis of these allergies remains uncertain. In order to determine whether eosinophils are involved in these GI-allergies or not, we measured eosinophil-derived neurotoxin (EDN) levels in stool samples from patients with various GI-allergies. METHODS: Stool samples were obtained from patients with GI-allergies (n542) or healthy infants (n5230). Stool sample extracts were prepared using sample preparation kits (Roche) and the EDN levels in the stool sample extracts were measured by ELISA. RESULTS: The EDN levels in the stool sample extracts from patients with GI-allergies were significantly higher than those from healthy infant. In addition, the EDN levels in the stool sample extracts varied among GIallergies; the EDN levels in the stool sample extract from patients who exhibited bloody stools were higher than those without bloody stools. CONCLUSIONS: Our results suggest that eosinophils are likely to be involved in the pathogenesis of GI-allergies, in particular, those with bloody stools. Our results also imply the involvement of Th2-type immune responses in the pathogenesis of GI-allergies through activation of eosinophils but not IgE synthesis.
354
Dynamics of Eosinophils in Non-IgE-mediated Gastrointestinal Food Allergies in Neonates and Infants, differences between 4 Clusters I. Nomura, H. Morita, T. Shoda, K. Arai, N. Ito, A. Nakazawa, Y. Ohya, H. Saito, K. Matsumoto; National Center for Child Health and Development, Tokyo, JAPAN. RATIONALE: Non-IgE-mediated gastrointestinal food allergies are consisted from several syndromes; FPIES, proctocolitis, enteropathy and allergic eosinophilic gastroenteritis (AEG). AEG and enteropathy can be diagnosed only by histological examination. To establish effective initial diagnosis of whole patients at first presentation, we tried to classify those by simple clinical variables and then, 4 clusters were derived (JACI 2011:127:685-688). In this paper, we tried to make comparison of eosinophils, important biological marker in GI allergy, between those 4 clusters. METHODS: Patients (total 300) were registered to database of our research group from 2007 to 2011. Clusters were defined according to presence of vomiting and/or bloody stool in the first 0-2 months after onset; Cluster 1 (vomiting +/ bloody stool +), Cluster 2 (+/-), Cluster 3 (-/-), Cluster 4 (-/+). Gastrointestinal histology; thirteen patients were examined their entire GI tract by fiberscope. Fecal eosinophils; mucous part of the feces was taken and stained with Hansel’s staining kit. RESULTS: Circulating eosinophil counts; highest value was seen in Cluster 3 (p<0.01), 14% of whole patients showed more than 30% of circulating eosinophils. Gastrointestinal histology; in Cluster 3 and 4, marked infiltration of eosinophils were admitted. In Cluster 3, inflammation was spread over entire GI tract but epithelial damage was minimal. Fecal eosinophils; high in Cluster 1 and 4, while Cluster 3 showed within normal limit. Data was confirmed by determination of fecal eosinophilderived neurotoxin. CONCLUSIONS: Difference in dynamics of eosinophils among 4 clusters was clarified. It might be very beneficial to conduct diagnosis and treatment considering this dynamics.
355
Our 3-year Experience with Patch Testing for Eosinophilic Esophagitis C. S. Bauer1, A. R. Lamba1, H. P. Zimmerman1, R. G. Hoffmann1, M. Dasgupta1, R. J. Noel1, M. B. Levy2, M. Vasudev1; 1Medical College of Wisconsin, Milwaukee, WI, 2Assaf Harofe Medical Center, Tzrifin, ISRAEL. RATIONALE: Treatment of eosinophilic esophagitis (EoE) consists of pharmacotherapy and/or dietary therapy. Food atopy patch testing (APT) for EoE is not standardized and its clinical utility is controversial. We hypothesize that our APT method is valuable adjunctive therapy to aid in treatment of pediatric EoE. METHODS: Sixty-two patients (male549, mean age58.8 years) with eosinophils on esophageal biopsy (mean proximally538/HPF, distally541/HPF) who were evaluated by food APT between 01/2008-02/ 2011 were retrospectively identified by ICD-9 codes. APT was performed using powdered food, sterile water, and glycerin in a 12-food EoE panel. Biopsies, food-allergy tests [serum IgE (sIgE), percutaneous skin test (PST), APT], and swallowed steroid (SS) usage were recorded. _1 food in 74%. For APT, RESULTS: Combined APT and PST identified > 53% (n533/62) were positive [peanut (n513/55, 24%), corn (n514/61, 23%), and chicken (n511/58, 19%)], for PST, 57% (n535/61; most commonly, peanut, wheat, and corn), and for sIgE, 67% (n512/18). Eleven patients had negative PST, but positive APT. Those with surveillance biopsies (n539) had 2 on average; 39% of surveillance biopsies (n531/79) were on guided-elimination (GE) from food-allergy testing. Those on SS (n512/39, 30.8%), SS and GE (n512/39, 30.8%), and GE (10.2%, n54/39) had 62% (proximally) and 69% (distally) decrease in mean eosinophils/HPF on first surveillance biopsy after APT compared to diagnostic biopsy. 56% (n510/18) of those on APT-based GE had <15/ eosinophils/HPF on first surveillance biopsy, though 70% of those (n57/ 10) were concomitantly on PST-based GE and SS. CONCLUSIONS: Our APT method with subsequent food elimination aids in the complex treatment of pediatric EoE.
SUNDAY
352