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Increased epithelial permeability and the generation of inflammatory cytokines in response to proteolytic activity in fruit
J ALLERGY CLIN IMMUNOL VOLUME 111, NUMBER 2
choose if: (1) the infant was well, and (2) the infant had atopic dermatitis. RESULTS: 78% of pediatricians (39/50), 100% of pediatric residents (26/26), and 80% of international experts (8/10) replied. If the infant was well, pediatricians chose CM (30.8%), soy (30.8%), and whey (33.3%) with similar frequency, 80.8% of residents chose CM, and experts chose CM (37.5%) or casein (50.0%), with one choosing soy. If the infant had atopic dermatitis, fewer pediatricians chose CM (12.8%), while more chose soy (38.5%) or casein (23.1%). Fewer residents chose CM (46.2%), and experts largely chose casein (62.5%). CONCLUSIONS: Our survey demonstrates a lack of consensus among pediatricians and experts for choice of formula. We hypothesize that cost may be an important consideration, More data are required to define the best approach for infants with a strong family history of cow's milk allergy.
Abstracts
S105
rechallenge (mean dose equivalence - 23 times less peanut). RAST inhibition showed that neither recipe altered epitope recognition but the higher fat recipe required twice as much peanut to cause 50% inhibition. ELISA detected far lower levels of peanut on the 31.5% recipe (220,000 ppm) than in the 22.9% fat recipe (990,000 ppm). CONCLUSIONS: These observations show that the fat content of a challenge vehicle has a profound effect on the reaction experienced after allergen ingestion. This is another factor to be considered in assessing the risk of certain foods to food allergic consumers and adds another dimension to clinical, research and regulatory practice.
Funding: Food Standards Agency
142
TransglutamiinnaseDi ChildrenN~ sease Foolproof in Diagnosis of Celiac
Funding: Self-funded
140 Inflammatmy Increased Epithelial Permeability and the Generation of Cytokinesin Responseto ProteolyticActivity in Fruit M. G. Buckley, A. F. Walls; Immunopharmacology Group, University of Southampton, Southampton, UNITED KINGDOM. RATIONALE: To investigate the hypothesis that constituents of certain tbods may interact directly with the epithelium to exacerbate or trigger allergic food reactions, we examined the potential of extracts of various fruits to alter epithelial permeability and induce cytokine production. METHODS: Chopstick electrodes were used to measure the trans-epithelial resistance (TER) of CACO-2 colonic epithelial cell monolayers as an inverse measure of permeability. Production of cytokine mRNA by 16HBE epithelial cells was determined by real time PCR. RESULTS: Pineapple extract significantly reduced TER 8-24 hours after application. Similar decreases were achieved using stem bromelain or the papaya enzyme papain. Heat treatment or the application of cysteine protease inhibitor E-64 could abrogate the effects of pineapple extract. Extracts of kiwi fruit, peach and mango had little effect on TER measurements. Pineapple extract caused a concentration-dependent up-regulation of GM-CSF mRNA, which increased 5.4-fold when 3 p.g/ml extract was applied. Treatment with E-64 abolished this increase, suggesting the involvement of a cysteine protease. Effects on IL-8 gene transcription were less pronounced, with a small, but statistically significant increase to 140% of control values following treatment with pineapple extract (P < 0.05). Mango extract increased GM-CSF mRNA to 2.2-fold control levels. CONCLUSIONS: Proteases in certain fruits may impair the integrity of the epithelial barrier, and interact directly with epithelial cells to stimulate the production of inflammatory cytokines.
Funding: Food Standards Agency ( UK Government)
141 Affect Presentation of Allergen in Different Food Preparations May the Severity of the Allergic Reaction J. O. HourihanO, K. E. C. G r i m s h a w I, R. M. King 2, J. M. Nordlee 3, S. L, Hefle 3, J. O. Warnert; 1University of Southampton, Southampton+ UNITED KINGDOM, 2Southampton University Hospital Trust, Wellcome Trust Clinical Research Facility, Southampton, UNITED KINGDOM, 3University of Nebraska-Lincoln, Lincoln, NE. RATIONALE: To determine whether reaction severity as observed in peanut allergic subjects undergoing double blind food challenge (DBFC) was due to differences in composition of the challenge vehicle. METHODS: An ongoing study into peanut allergy involves DBFC. Each challenge consists of eleven active doses of increasing magnitude randomly interspersed with tour placebo doses. Two different peanut recipes have been used, consisting of the same ingredients; the only difference being the percentage fat content (22.9% compared to 31.5%). Commercial ELISA and RAST inhibition studies were carried out to determine whether levels of detectable peanut differed between the two recipes. RESULTS: Four subjects were rechallenged with the 22.9% fat recipe after reacting more severely than anticipated to the 31.5% recipe. Three of the four subjects reacted to much smaller doses of peanut protein on
Z. Ko Beams, J. D. Lloyd-Still, R. Sandier, R. McConnie, S. Jakate; Rush Medical College, Chicago, IL. We reviewed 23 patients with Celiac Disease (CD) confirmed by improved health on Gluten Free Diet (GFD) to evaluate methods of diagnosing CD. We obtained the charts of children with confirmed CD. Patients were included in the study based on history, growth characteristics, laboratory tests, intestinal biopsy, and response to GFD. We reviewed the charts for results of tissue Transglutaminase (tTG), Anti-Gliadin Antibodies, Quantitative Immunoglobulins, intestinal biopsy, growth rate, and response to GFD. We found that of the 23 patients with CD confirmed by abnormal intestinal biopsy and improved health on GFD, only 12 (52%) had an abnormal tTG. This calls into question the utility of tTG as a screening and diagnostic tool for CD. The tTG test may be neither as specific or as sensitive as is currently thought. We conclude that improved health after GFD, combined with abnormal intestinal biopsy results and other clinical indicators, are more useful for identifying CD in children than an abnormal tTG. In the future, a double-blinded placebo controlled study could be used to further clarify the response to GFD and correlation with abnormal tTG.
Funding: Self-funded
143 the Comparison of Allergic Sensitization/Response to Soybean in Neonatal Swine Peanut Allergy Model R. M. Helm ].2, G. T. Furuta 3, J. S. Stanley ],2, C. M. West ],2, G. Cockrell j,2, C. Connaughton 1,2, A. W. Burksl,2; tPediatrics, University of Arkansas for Medical Sciences, Little Rock, AR, 2Arkansas Children's Hospital, Little Rock, AR, 3Harvard Medical School, Boston, MA. The neonatal swine food allergy model under investigation in our laboratory is being used to determine the immunoregulatory mechanism of clinically relevant gastrointestinal immediate hypersensitivity, develop predictive roles Ior food and novel proteins in immune sensitization/allergic responses, and to offer therapeutic options for treatment of food allergy. RATIONALE: To determine and compare the relative sensitivity and allergic response of soybean in the swine peanut allergy model. METHODS: Neonatal piglets reared on a soybean/peanut-free diet were intraperitoneally sensitized to 500 or 1000 p.g of soybean extract plus cholera toxin. Additional piglets were also sensitized 500 I.tg peanut extract plus cholera toxin and carrier control plus cholera toxin. Oral challenge by gastric intubation was performed with 10-15 grams of soybean or peanut meal. Skin testing was performed with soybean and peanut extracts. RESULTS: Oral challenged soybean-sensitized piglets responded with mild to moderate responses (whole body rashes, lethargy) compared to more severe responses (vomiting, diarrhea) in peanut sensitized animals. Skin test confirmed sensitization to the respective allergen with little or no response to the non-sensitizing allergen. CONCLUSIONS: The swine peanut allergy model can be used for additional food allergen sensitization. The clinical symptoms and allergic response to soybean correlates with that seen in food allergic responses in soybean/peanut sensitive individuals.
Funding: USDA
choose if: (1) the infant was well, and (2) the infant had atopic dermatitis. RESULTS: 78% of pediatricians (39/50), 100% of pediatric residents (26/26), and 80% of international experts (8/10) replied. If the infant was well, pediatricians chose CM (30.8%), soy (30.8%), and whey (33.3%) with similar frequency, 80.8% of residents chose CM, and experts chose CM (37.5%) or casein (50.0%), with one choosing soy. If the infant had atopic dermatitis, fewer pediatricians chose CM (12.8%), while more chose soy (38.5%) or casein (23.1%). Fewer residents chose CM (46.2%), and experts largely chose casein (62.5%). CONCLUSIONS: Our survey demonstrates a lack of consensus among pediatricians and experts for choice of formula. We hypothesize that cost may be an important consideration, More data are required to define the best approach for infants with a strong family history of cow's milk allergy.
Abstracts
S105
rechallenge (mean dose equivalence - 23 times less peanut). RAST inhibition showed that neither recipe altered epitope recognition but the higher fat recipe required twice as much peanut to cause 50% inhibition. ELISA detected far lower levels of peanut on the 31.5% recipe (220,000 ppm) than in the 22.9% fat recipe (990,000 ppm). CONCLUSIONS: These observations show that the fat content of a challenge vehicle has a profound effect on the reaction experienced after allergen ingestion. This is another factor to be considered in assessing the risk of certain foods to food allergic consumers and adds another dimension to clinical, research and regulatory practice.
Funding: Food Standards Agency
142
TransglutamiinnaseDi ChildrenN~ sease Foolproof in Diagnosis of Celiac
Funding: Self-funded
140 Inflammatmy Increased Epithelial Permeability and the Generation of Cytokinesin Responseto ProteolyticActivity in Fruit M. G. Buckley, A. F. Walls; Immunopharmacology Group, University of Southampton, Southampton, UNITED KINGDOM. RATIONALE: To investigate the hypothesis that constituents of certain tbods may interact directly with the epithelium to exacerbate or trigger allergic food reactions, we examined the potential of extracts of various fruits to alter epithelial permeability and induce cytokine production. METHODS: Chopstick electrodes were used to measure the trans-epithelial resistance (TER) of CACO-2 colonic epithelial cell monolayers as an inverse measure of permeability. Production of cytokine mRNA by 16HBE epithelial cells was determined by real time PCR. RESULTS: Pineapple extract significantly reduced TER 8-24 hours after application. Similar decreases were achieved using stem bromelain or the papaya enzyme papain. Heat treatment or the application of cysteine protease inhibitor E-64 could abrogate the effects of pineapple extract. Extracts of kiwi fruit, peach and mango had little effect on TER measurements. Pineapple extract caused a concentration-dependent up-regulation of GM-CSF mRNA, which increased 5.4-fold when 3 p.g/ml extract was applied. Treatment with E-64 abolished this increase, suggesting the involvement of a cysteine protease. Effects on IL-8 gene transcription were less pronounced, with a small, but statistically significant increase to 140% of control values following treatment with pineapple extract (P < 0.05). Mango extract increased GM-CSF mRNA to 2.2-fold control levels. CONCLUSIONS: Proteases in certain fruits may impair the integrity of the epithelial barrier, and interact directly with epithelial cells to stimulate the production of inflammatory cytokines.
Funding: Food Standards Agency ( UK Government)
141 Affect Presentation of Allergen in Different Food Preparations May the Severity of the Allergic Reaction J. O. HourihanO, K. E. C. G r i m s h a w I, R. M. King 2, J. M. Nordlee 3, S. L, Hefle 3, J. O. Warnert; 1University of Southampton, Southampton+ UNITED KINGDOM, 2Southampton University Hospital Trust, Wellcome Trust Clinical Research Facility, Southampton, UNITED KINGDOM, 3University of Nebraska-Lincoln, Lincoln, NE. RATIONALE: To determine whether reaction severity as observed in peanut allergic subjects undergoing double blind food challenge (DBFC) was due to differences in composition of the challenge vehicle. METHODS: An ongoing study into peanut allergy involves DBFC. Each challenge consists of eleven active doses of increasing magnitude randomly interspersed with tour placebo doses. Two different peanut recipes have been used, consisting of the same ingredients; the only difference being the percentage fat content (22.9% compared to 31.5%). Commercial ELISA and RAST inhibition studies were carried out to determine whether levels of detectable peanut differed between the two recipes. RESULTS: Four subjects were rechallenged with the 22.9% fat recipe after reacting more severely than anticipated to the 31.5% recipe. Three of the four subjects reacted to much smaller doses of peanut protein on
Z. Ko Beams, J. D. Lloyd-Still, R. Sandier, R. McConnie, S. Jakate; Rush Medical College, Chicago, IL. We reviewed 23 patients with Celiac Disease (CD) confirmed by improved health on Gluten Free Diet (GFD) to evaluate methods of diagnosing CD. We obtained the charts of children with confirmed CD. Patients were included in the study based on history, growth characteristics, laboratory tests, intestinal biopsy, and response to GFD. We reviewed the charts for results of tissue Transglutaminase (tTG), Anti-Gliadin Antibodies, Quantitative Immunoglobulins, intestinal biopsy, growth rate, and response to GFD. We found that of the 23 patients with CD confirmed by abnormal intestinal biopsy and improved health on GFD, only 12 (52%) had an abnormal tTG. This calls into question the utility of tTG as a screening and diagnostic tool for CD. The tTG test may be neither as specific or as sensitive as is currently thought. We conclude that improved health after GFD, combined with abnormal intestinal biopsy results and other clinical indicators, are more useful for identifying CD in children than an abnormal tTG. In the future, a double-blinded placebo controlled study could be used to further clarify the response to GFD and correlation with abnormal tTG.
Funding: Self-funded
143 the Comparison of Allergic Sensitization/Response to Soybean in Neonatal Swine Peanut Allergy Model R. M. Helm ].2, G. T. Furuta 3, J. S. Stanley ],2, C. M. West ],2, G. Cockrell j,2, C. Connaughton 1,2, A. W. Burksl,2; tPediatrics, University of Arkansas for Medical Sciences, Little Rock, AR, 2Arkansas Children's Hospital, Little Rock, AR, 3Harvard Medical School, Boston, MA. The neonatal swine food allergy model under investigation in our laboratory is being used to determine the immunoregulatory mechanism of clinically relevant gastrointestinal immediate hypersensitivity, develop predictive roles Ior food and novel proteins in immune sensitization/allergic responses, and to offer therapeutic options for treatment of food allergy. RATIONALE: To determine and compare the relative sensitivity and allergic response of soybean in the swine peanut allergy model. METHODS: Neonatal piglets reared on a soybean/peanut-free diet were intraperitoneally sensitized to 500 or 1000 p.g of soybean extract plus cholera toxin. Additional piglets were also sensitized 500 I.tg peanut extract plus cholera toxin and carrier control plus cholera toxin. Oral challenge by gastric intubation was performed with 10-15 grams of soybean or peanut meal. Skin testing was performed with soybean and peanut extracts. RESULTS: Oral challenged soybean-sensitized piglets responded with mild to moderate responses (whole body rashes, lethargy) compared to more severe responses (vomiting, diarrhea) in peanut sensitized animals. Skin test confirmed sensitization to the respective allergen with little or no response to the non-sensitizing allergen. CONCLUSIONS: The swine peanut allergy model can be used for additional food allergen sensitization. The clinical symptoms and allergic response to soybean correlates with that seen in food allergic responses in soybean/peanut sensitive individuals.
Funding: USDA