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Advisory Labeling and Contamination Levels for Egg, Milk and Peanut
S176 Abstracts
672
Advisory Labeling and Contamination Levels for Egg, Milk and Peanut L. Ford1, S. Taylor2, L. Niemann2, D. Lambrecht2, T. Slotkin3, R. Pacenza3, S. H. Sicherer1; 1Mt. Sinai School of Medicine, New York, NY, 2 Food Allergy Research & Resource Program, University of Nebraska, Lincoln, NE, 3Food Allergy Initiative, New York, NY. RATIONALE: Product label advisory warnings ("may contain") are voluntary and unregulated. The risks of allergen contamination remain substantially unexplored. METHODS: Allergen assays (Neogen VeratoxÒ ELISA) were performed on a convenience sample of 399 food products selected for the presence of advisory labeling for egg, milk or peanut, or for being similar to such products but without a warning. Products labeled as utilizing Good Manufacturing Procedures for allergens (n 5 26) were considered to be among those with no allergens disclosed. RESULTS: Assays detected the three allergens as follows: egg in 1/57 (1.8%) products with egg advisory warnings and 3/117 (2.6%) with no allergen declared (concentration, 3-8 ppm); milk in 6/59 (10.2%) products with milk advisory warnings and 4/134 (3.0%) with no allergen declared (4-222 ppm); peanut in 5/112 (4.5%) products with advisory warnings and 0/120 (0%) with no allergen declared (5-161 ppm). Overall, 7/19 contained below 10 ppm. Peanut and milk, but not egg, were significantly (p < 0.05) more likely to be found in products with a warning statement than in products without. Milk, but not egg or peanut, was significantly more likely to be found in products from small, vs. large, manufacturers. The 7 contaminated products without warnings (egg, 3-8 ppm; milk, 17-124 ppm) were from 5 companies; only one was from a large company (egg, 7 ppm). CONCLUSIONS: Overall, 5.3% of advisory labeled products and 1.9% without allergen declared contained the allergen. Contaminated foods without advisory labeling were primarily from small manufacturers. The clinical impact of the degree of contamination remains to be determined.
673
MONDAY
Investigation of the Safety of the Food Allergy Herbal Formula (FAHF-2) in Patients with Food Allergy-Phase 1 J. Wang, S. A. Noone, S. Patil, H. A. Sampson, X. Li; Mount Sinai Medical Center, New York, NY. RATIONALE: Chinese herbal medicine is increasingly attracting interest in Western countries as an alternative therapy for a variety of diseases, including allergies and asthma. Preclinical studies have demonstrated that the Food Allergy Herbal Formula (FAHF-2), consisting of 9 herbs, completely protected a murine model against peanut-induced anaphylaxis up to 6 months following therapy. We investigated the safety of FAHF-2 in food allergic patients in this phase I study (IND 77 468). METHODS: Patients 12-45 years of age, who have a history of peanut, tree nut, fish or shellfish allergy were eligible for this randomized, double-blind, placebo-controlled dose escalation phase I trial. Groups of 6 individuals received FAHF-2 at doses of 2.2 grams (4 tablets), 3.3 grams (6 tablets) or 6.6 grams (12 tablets) three times a day for 7 days. Subjects continued allergen avoidance for the duration of the study. Vital signs, physical examination, laboratory data, electrocardiographic data, and pulmonary function tests were monitored to assess for adverse events. The dose increase was allowed after a careful review of the safety data of the lower dose group. RESULTS: To date, 12 subjects have been enrolled and randomized to receive FAHF-2 or placebo. All have successfully completed treatment. No significant differences in vital signs, laboratory data, electrocardiographic findings or pulmonary function test data were observed before and after treatment. Two patients reported mild gastrointestinal symptoms (one FAHF-2 and one placebo) which resolved without intervention. CONCLUSIONS: FAHF-2 appears to be safe and well tolerated by food allergic patients at the low and medium doses.
J ALLERGY CLIN IMMUNOL FEBRUARY 2009
674
Gly m 5 is a Major Soybean Allergen in European Children: Comparative Investigation of Linear B-cell Epitopes of Gly m 5 and Ara h 1, the Homologous Peanut Allergen Y. Kuehne1, B. Niggemann2, B. K. Ballmer-Weber3, J. Scibilia4, C. Bindslev-Jensen5, O. Wackermann1, G. Reese1, S. Vieths1, T. Holzhauser1; 1 Paul-Ehrlich-Institut, Langen, Germany, 2Charite University Medical Centre, Berlin, Germany, 3Universital Hospital, Zurich, Switzerland, 4Niguarda Ca Granda Hospital, Milan, Italy, 5University Hospital, Odense, Denmark. RATIONALE: Soybean-allergic subjects are frequently cosensitized to peanut. Published data on soybean allergens and epitopes are incomplete. Thus, we investigated the relevance of Gly m 5 (beta-conglycinin), and its linear B-cell epitopes, in comparison to Ara h 1, the homologous vicilin in peanut. METHODS: Fifty-nine subjects with clinically confirmed soybean allergy were enrolled. Inclusion critera were a positive DBPCFC or convincing history of anaphylaxis to soybean. With overlapping 15-mer peptides covering the complete protein sequence, Gly m 5 reactive sera were mapped for linear IgE-binding epitopes of Ara h 1 and of the three beta-conglycinin subunits: Gly m 5.01, Gly m 5.02, and Gly m 5.03. RESULTS: 44% (26/59) of all study subjects, 78% (18/23) of the soy-allergic children and 71% (5/7) of subjects with soybean anaphylaxis showed specific IgE to Gly m 5. 65% (17/26) displayed IgE-binding to linear Gly m 5 epitopes and 85% (22/26) to linear Ara h 1 epitopes, respectively. Two adjacent major IgE-reactive surface areas of Gly m 5 were identified by 3D modelling. The IgE epitope areas of all subunits were similar. Ara h 1 epitopes recognized by soybean-allergic subjects with a history of peanut allergy were located at protein surface areas other than those previously described for peanut-allergic subjects. CONCLUSIONS: Gly m 5 is a major allergen in soybean-allergic children and potentially a marker for severe soy-related allergic reactions. The observed differences in IgE epitope pattern between soybean and peanutallergic subjects may be due to differences in cosensitization or geographic origin.
675
Identification of T cell Epitopes for Ara h 1 in Peanut Allergic Individuals Using HLA Class II Tetramers K. A. Hetherington1, D. M. Robinson2, E. A. James3, W. W. Kwok3; 1 University of Washington, Seattle, WA, 2Virginia Mason Medical Center, Seattle, WA, 3Benaroya Research Institute, Seattle, WA. RATIONALE: Peptide immunotherapy and diagnostic allergy tests could be made more safe and effective through the identification of haplotype specific T cell epitopes. Our goal is to identify MHC class II restricted epitopes for the major peanut allergen, Ara h 1, using novel MHC class II tetramer reagents. METHODS: We used MHC class II tetramers and Tetramer Guided Epitope Mapping to identify the antigenic peptides for the peanut allergen protein Ara h 1. In initial studies, peripheral blood mononucleated cells were isolated from one peanut-allergic individual with an HLADRB1*0401 haplotype and one peanut-allergic individual with an HLADRB1*0404 haplotype. T cells were stimulated with pools of 20 amino acid peptides representing the entire sequence of the Ara h 1 and stained with corresponding tetramer reagents after in vitro amplification. RESULTS: In the DR0401 peanut-allergic patient, two DR0401 restricted antigenic peptides were found: Ara h 1 (329-348) and Ara h 1 (577-596). In the DR0404 peanut-allergic patient two DR0404 restricted antigenic peptides were found: the same Ara h 1 (329-348) peptide and Ara h 1 (449468). CONCLUSIONS: HLA restricted antigenic peptides for Ara h 1 were identified for 2 peanut allergic individuals with haplotypes HLADRB1*0401 and HLA-DRB1*0404. These findings demonstrate that MHC class II tetramers can provide a rapid and effective approach for the identification of HLA restricted epitopes in peanut allergic subjects and suggest that some Ara h 1 epitopes are haplotype specific.
672
Advisory Labeling and Contamination Levels for Egg, Milk and Peanut L. Ford1, S. Taylor2, L. Niemann2, D. Lambrecht2, T. Slotkin3, R. Pacenza3, S. H. Sicherer1; 1Mt. Sinai School of Medicine, New York, NY, 2 Food Allergy Research & Resource Program, University of Nebraska, Lincoln, NE, 3Food Allergy Initiative, New York, NY. RATIONALE: Product label advisory warnings ("may contain") are voluntary and unregulated. The risks of allergen contamination remain substantially unexplored. METHODS: Allergen assays (Neogen VeratoxÒ ELISA) were performed on a convenience sample of 399 food products selected for the presence of advisory labeling for egg, milk or peanut, or for being similar to such products but without a warning. Products labeled as utilizing Good Manufacturing Procedures for allergens (n 5 26) were considered to be among those with no allergens disclosed. RESULTS: Assays detected the three allergens as follows: egg in 1/57 (1.8%) products with egg advisory warnings and 3/117 (2.6%) with no allergen declared (concentration, 3-8 ppm); milk in 6/59 (10.2%) products with milk advisory warnings and 4/134 (3.0%) with no allergen declared (4-222 ppm); peanut in 5/112 (4.5%) products with advisory warnings and 0/120 (0%) with no allergen declared (5-161 ppm). Overall, 7/19 contained below 10 ppm. Peanut and milk, but not egg, were significantly (p < 0.05) more likely to be found in products with a warning statement than in products without. Milk, but not egg or peanut, was significantly more likely to be found in products from small, vs. large, manufacturers. The 7 contaminated products without warnings (egg, 3-8 ppm; milk, 17-124 ppm) were from 5 companies; only one was from a large company (egg, 7 ppm). CONCLUSIONS: Overall, 5.3% of advisory labeled products and 1.9% without allergen declared contained the allergen. Contaminated foods without advisory labeling were primarily from small manufacturers. The clinical impact of the degree of contamination remains to be determined.
673
MONDAY
Investigation of the Safety of the Food Allergy Herbal Formula (FAHF-2) in Patients with Food Allergy-Phase 1 J. Wang, S. A. Noone, S. Patil, H. A. Sampson, X. Li; Mount Sinai Medical Center, New York, NY. RATIONALE: Chinese herbal medicine is increasingly attracting interest in Western countries as an alternative therapy for a variety of diseases, including allergies and asthma. Preclinical studies have demonstrated that the Food Allergy Herbal Formula (FAHF-2), consisting of 9 herbs, completely protected a murine model against peanut-induced anaphylaxis up to 6 months following therapy. We investigated the safety of FAHF-2 in food allergic patients in this phase I study (IND 77 468). METHODS: Patients 12-45 years of age, who have a history of peanut, tree nut, fish or shellfish allergy were eligible for this randomized, double-blind, placebo-controlled dose escalation phase I trial. Groups of 6 individuals received FAHF-2 at doses of 2.2 grams (4 tablets), 3.3 grams (6 tablets) or 6.6 grams (12 tablets) three times a day for 7 days. Subjects continued allergen avoidance for the duration of the study. Vital signs, physical examination, laboratory data, electrocardiographic data, and pulmonary function tests were monitored to assess for adverse events. The dose increase was allowed after a careful review of the safety data of the lower dose group. RESULTS: To date, 12 subjects have been enrolled and randomized to receive FAHF-2 or placebo. All have successfully completed treatment. No significant differences in vital signs, laboratory data, electrocardiographic findings or pulmonary function test data were observed before and after treatment. Two patients reported mild gastrointestinal symptoms (one FAHF-2 and one placebo) which resolved without intervention. CONCLUSIONS: FAHF-2 appears to be safe and well tolerated by food allergic patients at the low and medium doses.
J ALLERGY CLIN IMMUNOL FEBRUARY 2009
674
Gly m 5 is a Major Soybean Allergen in European Children: Comparative Investigation of Linear B-cell Epitopes of Gly m 5 and Ara h 1, the Homologous Peanut Allergen Y. Kuehne1, B. Niggemann2, B. K. Ballmer-Weber3, J. Scibilia4, C. Bindslev-Jensen5, O. Wackermann1, G. Reese1, S. Vieths1, T. Holzhauser1; 1 Paul-Ehrlich-Institut, Langen, Germany, 2Charite University Medical Centre, Berlin, Germany, 3Universital Hospital, Zurich, Switzerland, 4Niguarda Ca Granda Hospital, Milan, Italy, 5University Hospital, Odense, Denmark. RATIONALE: Soybean-allergic subjects are frequently cosensitized to peanut. Published data on soybean allergens and epitopes are incomplete. Thus, we investigated the relevance of Gly m 5 (beta-conglycinin), and its linear B-cell epitopes, in comparison to Ara h 1, the homologous vicilin in peanut. METHODS: Fifty-nine subjects with clinically confirmed soybean allergy were enrolled. Inclusion critera were a positive DBPCFC or convincing history of anaphylaxis to soybean. With overlapping 15-mer peptides covering the complete protein sequence, Gly m 5 reactive sera were mapped for linear IgE-binding epitopes of Ara h 1 and of the three beta-conglycinin subunits: Gly m 5.01, Gly m 5.02, and Gly m 5.03. RESULTS: 44% (26/59) of all study subjects, 78% (18/23) of the soy-allergic children and 71% (5/7) of subjects with soybean anaphylaxis showed specific IgE to Gly m 5. 65% (17/26) displayed IgE-binding to linear Gly m 5 epitopes and 85% (22/26) to linear Ara h 1 epitopes, respectively. Two adjacent major IgE-reactive surface areas of Gly m 5 were identified by 3D modelling. The IgE epitope areas of all subunits were similar. Ara h 1 epitopes recognized by soybean-allergic subjects with a history of peanut allergy were located at protein surface areas other than those previously described for peanut-allergic subjects. CONCLUSIONS: Gly m 5 is a major allergen in soybean-allergic children and potentially a marker for severe soy-related allergic reactions. The observed differences in IgE epitope pattern between soybean and peanutallergic subjects may be due to differences in cosensitization or geographic origin.
675
Identification of T cell Epitopes for Ara h 1 in Peanut Allergic Individuals Using HLA Class II Tetramers K. A. Hetherington1, D. M. Robinson2, E. A. James3, W. W. Kwok3; 1 University of Washington, Seattle, WA, 2Virginia Mason Medical Center, Seattle, WA, 3Benaroya Research Institute, Seattle, WA. RATIONALE: Peptide immunotherapy and diagnostic allergy tests could be made more safe and effective through the identification of haplotype specific T cell epitopes. Our goal is to identify MHC class II restricted epitopes for the major peanut allergen, Ara h 1, using novel MHC class II tetramer reagents. METHODS: We used MHC class II tetramers and Tetramer Guided Epitope Mapping to identify the antigenic peptides for the peanut allergen protein Ara h 1. In initial studies, peripheral blood mononucleated cells were isolated from one peanut-allergic individual with an HLADRB1*0401 haplotype and one peanut-allergic individual with an HLADRB1*0404 haplotype. T cells were stimulated with pools of 20 amino acid peptides representing the entire sequence of the Ara h 1 and stained with corresponding tetramer reagents after in vitro amplification. RESULTS: In the DR0401 peanut-allergic patient, two DR0401 restricted antigenic peptides were found: Ara h 1 (329-348) and Ara h 1 (577-596). In the DR0404 peanut-allergic patient two DR0404 restricted antigenic peptides were found: the same Ara h 1 (329-348) peptide and Ara h 1 (449468). CONCLUSIONS: HLA restricted antigenic peptides for Ara h 1 were identified for 2 peanut allergic individuals with haplotypes HLADRB1*0401 and HLA-DRB1*0404. These findings demonstrate that MHC class II tetramers can provide a rapid and effective approach for the identification of HLA restricted epitopes in peanut allergic subjects and suggest that some Ara h 1 epitopes are haplotype specific.