- Email: [email protected]
Usefulness of Diagnostic Tests in Cows' Milk Allergy
185
Usefulness of Diagnostic Tests in Cows’ Milk Allergy
I. CERECEDO1, M. DIEGUEZ1, A. MURIEL2, A. GARCIA BALDA1, S. TERRADOS1, M. SANCHEZ CANO1, B. DE LA HOZ1; 1ALLERGY, RAMON Y CAJAL, MADRID, SPAIN, 2Unidad De Bioestadistica, RAMON Y CAJAL, MADRID, SPAIN. RATIONALE: Different cutoff values for skin prik test (SPT) and specific IgE levels have been previously given to predict a positive food challenge test in milk (CM) allergy. METHODS: A transversal study including CM allergic children. After six months of milk-excluding-diet a complete anamnesis, SPT and specific IgE with CM and fractions were determinated. Simpled-blind controlled food challenge (SBCFC) were performed in children with specific IgE lower than 2.5 KU/l. The others continued with milk-excluding-diet and after 6 months were asked for any transgression in diet. We considered this transgresions as a food oral chalenge test at home. RESULTS: 162 children were included. Only 85 children had specific IgE lower than 2.5KU/l and a SBCFC was performed. 54 tolerated (63.5%). 77 continued without CM.13 had a symptomatic transgresion (possitive chalenge). Best cutoff points for SPT to milk and fractions were 3mm, except for alfalactoalbumine (6mm). The cutoff value 2,5 Ku/l of CM specific-IgE in our study had a specificty of 79,2% and a negative predictive value of 60%. Differences in accuracy were obteined to SPT and milk specific IgE between older and younger than one year. CM SPT ROC area was 0.54 in <1 year and 0.80 in >1year, and milk specific IgE ROC area was 0.40 in <1 year and 0.75 in >1 year. CONCLUSIONS: The accuracy of previously given 2,5 Ku/l cutoff point for specific IgE in the patient management in clinical practice was similar to published studies in clinical research conditionts. Test accuracy was higher in children older than 1 year. Cas S 8, the Lipid Transfer Protein from Chestnut Seeds, is a Major Allergen in Chestnut Allergic Patients without Associated Latex Allergy, but not in those with the Latex-Fruit Syndrome C. Blanco1, R. Sánchez-Monge2, M. Recas2, G. López-Torrejón2, J. Cumplido1, J. Figueroa1, T. Carrillo1, G. Salcedo2; 1Hospital Universitario Dr. Negrín, Las Palmas de GC, SPAIN, 2E.T.S. Ingenieros Agrónomos, Madrid, SPAIN. RATIONALE: The allergens involved in chestnut allergy without associated latex hypersensitivity have not been studied. METHODS: Sera from 12 patients with chestnut but not latex allergy, as well as from 3 subjects with the latex fruit syndrome, were used. Chestnut lipid transfer protein (LTP) Cas s 8 was purified by gel-filtration chromatography followed by RP-HPLC, and characterized by N-terminal amino acid sequencing and MALDI analysis. Isolated Cas s 8, together with peach LTP Pru p 3, and avocado class I chitinase Pers a 1 (homologous to Cas s 5), were tested in immunodetection, ELISA and skin prick test (SPT). RESULTS: LTPs Cas s 8 and Pru p 3, but not chitinase Pers a 1, were recognized in IgE immunodetection assays using a serum pool from patients with chestnut but not latex allergy. The opposite was true when a pool of sera from patients with latex-fruit syndrome was analyzed. Specific IgE to Cas s 8 was found in 11/12 and 7/12 individual sera from chestnut but not latex allergic patients, tested by immunodetection and direct ELISA, respectively. All these sera had specific IgE to Pru p 3, whereas only one of them to Pers a 1. Cas s 8, as well as Pru p 3, inhibited 61% of the IgE binding to a chestnut crude extract in ELISA-inhibition assays. CONCLUSIONS: A different pattern of main allergens is shown in chestnut but not latex allergy, compared to the latex-fruit syndrome, being LTP the major allergen in the former, and class I chitinase in the later case. Funding: Instituto de Salud Carlos III (ISCiii RTIC G03/094 & C03/011)
186
Sensitization to Non-Specific Lipid Transfer Protein from Parietaria Judaica Pollen and Cross Sensitization to Foods ~as1, B. Bartolomé2; 1Unitat d’Al·lèrgia, M. Ibero1, M. Castillo1, M. Vin Hospital de Terrassa, Terrassa, SPAIN, 2I + D, Laboratorios BIAL-Aristegui, Bilbao, SPAIN. CASE REPORT: A 16-year-old boy, diagnosed of rhinoconjunctivitis and sensitizated to: dust, molds, cat epithelium, dog epithelium and pollens (Olea europea, Cupressus arizonica, Lolium perenne, Cynodon dactylon, Parietaria judaica and Plantago lanceolata), that suffered from anaphylaxis after eating chicken with potatoes and artichoke. Several months later the patient suffered from swelling of the lips with almonds. Later he has tolerated chicken and potatoes. METHODS AND RESULTS: Skin prick tests were positive to extracts from: artichoke (crude and cooked) and nuts. Specific IgE (EAST) were: Flower of Artichoke (flower+head-food): 8.6 kU/l; Stem of Artichoke: 3.6 kU/l; Artichoke (head-food): 2.9 kU/l; peanut: 26.6 kU/l and almonds: 2.23 kU/l.. SDS-PAGE Immunoblotting with artichoke extract showed IgE-binding bands at 14 kDa, 15,5 kDa and 17 kDa. Cross reactivity was studied between the artichoke extract and the extract from these pollens to which the patient was sensitizated. EAST inhibition using artichoke extract as solid phase showed an inhibition of 60% with P. judaica pollen extract. Immunoblotting with artichoke extract showed a complete inhibition of the IgE-binding when patient serum was preincubated with P. judaica pollen extract. Inhibition studies showed the artichoke IgEbinding proteins cross-react with the lipid transfer protein (LTP) from peach. CONCLUSIONS: In our knowledge it is the first case of oral allergy to artichoke. The clinical evolution of the patient and the in vitro test suggest an initial sensitization by respiratory tract to pollen from P. judaica, and a later cross sensitization by digestive tract to artichoke.
187
Analysis of Ag-specific Ige Antibody, Total Serum Ige and Blood Eosinophil in Food Allergic Children J. Kim1, Y. Kim2, H. Lee2; 1Department of Pediatrics, Catholic University Medical College, Uijongbu-city, REPUBLIC OF KOREA, 2Department of Clinical Pathology, Catholic University Medical College, Uijongbu-city, REPUBLIC OF KOREA. RATIONALE: Challenge test remain standard for diagnosis of food allergy, they are difficult to perform and laboratory tests are usually used. We evaluated the correlations of food Ag-specific IgE class (MAST CLA, ImmunoCAP100), total IgE class (MAST CLA), total serum IgE, and blood eosinophil count in food allergic children. METHODS: A total of 293 children who visited allergy clinic were tested by Korean food panel of MAST CLA. RESULTS: In 17 of 38 children, who show higher than MAST CLA Class 2 without Ag-specific IgE class, were tested by ImmunoCAP100 for egg white, milk, wheat, soy bean, peanut, cat epithelium, dog epithelium, D. farinae, and D. pteronyssinus. There was statistical difference(p<0.05) between total IgE class0-1/0 and total IgE class 4 in the number of Agspecific IgE. Total IgE class 4 show larger number of Ag-specific IgE class than total IgE class 0-1/0. But no statistical differences between total IgE class0-1/0 and total IgE class 1, 2 and 3. Although no statistical differences between total serum IgE and the number of Ag-specific IgE class, there were statistical differences(p<0.05) between total IgE class 4 and total IgE class 0-1/0, 1, 2, and 3 respectively. No differences between blood eosinophil count and number of Ag-specific IgE class, total IgE class, total serum IgE level. 6(35.3%) of the 17 children, was higher than MAST CLA Class 2 without Ag-specific IgE class, show positive reaction in ImmunoCAP100 test. CONCLUSIONS: Total IgE class 4 show larger number of Ag-specific IgE class than that of total IgE class 0-1/0.
188
SATURDAY
Abstracts S47
J ALLERGY CLIN IMMUNOL VOLUME 117, NUMBER 2
Usefulness of Diagnostic Tests in Cows’ Milk Allergy
I. CERECEDO1, M. DIEGUEZ1, A. MURIEL2, A. GARCIA BALDA1, S. TERRADOS1, M. SANCHEZ CANO1, B. DE LA HOZ1; 1ALLERGY, RAMON Y CAJAL, MADRID, SPAIN, 2Unidad De Bioestadistica, RAMON Y CAJAL, MADRID, SPAIN. RATIONALE: Different cutoff values for skin prik test (SPT) and specific IgE levels have been previously given to predict a positive food challenge test in milk (CM) allergy. METHODS: A transversal study including CM allergic children. After six months of milk-excluding-diet a complete anamnesis, SPT and specific IgE with CM and fractions were determinated. Simpled-blind controlled food challenge (SBCFC) were performed in children with specific IgE lower than 2.5 KU/l. The others continued with milk-excluding-diet and after 6 months were asked for any transgression in diet. We considered this transgresions as a food oral chalenge test at home. RESULTS: 162 children were included. Only 85 children had specific IgE lower than 2.5KU/l and a SBCFC was performed. 54 tolerated (63.5%). 77 continued without CM.13 had a symptomatic transgresion (possitive chalenge). Best cutoff points for SPT to milk and fractions were 3mm, except for alfalactoalbumine (6mm). The cutoff value 2,5 Ku/l of CM specific-IgE in our study had a specificty of 79,2% and a negative predictive value of 60%. Differences in accuracy were obteined to SPT and milk specific IgE between older and younger than one year. CM SPT ROC area was 0.54 in <1 year and 0.80 in >1year, and milk specific IgE ROC area was 0.40 in <1 year and 0.75 in >1 year. CONCLUSIONS: The accuracy of previously given 2,5 Ku/l cutoff point for specific IgE in the patient management in clinical practice was similar to published studies in clinical research conditionts. Test accuracy was higher in children older than 1 year. Cas S 8, the Lipid Transfer Protein from Chestnut Seeds, is a Major Allergen in Chestnut Allergic Patients without Associated Latex Allergy, but not in those with the Latex-Fruit Syndrome C. Blanco1, R. Sánchez-Monge2, M. Recas2, G. López-Torrejón2, J. Cumplido1, J. Figueroa1, T. Carrillo1, G. Salcedo2; 1Hospital Universitario Dr. Negrín, Las Palmas de GC, SPAIN, 2E.T.S. Ingenieros Agrónomos, Madrid, SPAIN. RATIONALE: The allergens involved in chestnut allergy without associated latex hypersensitivity have not been studied. METHODS: Sera from 12 patients with chestnut but not latex allergy, as well as from 3 subjects with the latex fruit syndrome, were used. Chestnut lipid transfer protein (LTP) Cas s 8 was purified by gel-filtration chromatography followed by RP-HPLC, and characterized by N-terminal amino acid sequencing and MALDI analysis. Isolated Cas s 8, together with peach LTP Pru p 3, and avocado class I chitinase Pers a 1 (homologous to Cas s 5), were tested in immunodetection, ELISA and skin prick test (SPT). RESULTS: LTPs Cas s 8 and Pru p 3, but not chitinase Pers a 1, were recognized in IgE immunodetection assays using a serum pool from patients with chestnut but not latex allergy. The opposite was true when a pool of sera from patients with latex-fruit syndrome was analyzed. Specific IgE to Cas s 8 was found in 11/12 and 7/12 individual sera from chestnut but not latex allergic patients, tested by immunodetection and direct ELISA, respectively. All these sera had specific IgE to Pru p 3, whereas only one of them to Pers a 1. Cas s 8, as well as Pru p 3, inhibited 61% of the IgE binding to a chestnut crude extract in ELISA-inhibition assays. CONCLUSIONS: A different pattern of main allergens is shown in chestnut but not latex allergy, compared to the latex-fruit syndrome, being LTP the major allergen in the former, and class I chitinase in the later case. Funding: Instituto de Salud Carlos III (ISCiii RTIC G03/094 & C03/011)
186
Sensitization to Non-Specific Lipid Transfer Protein from Parietaria Judaica Pollen and Cross Sensitization to Foods ~as1, B. Bartolomé2; 1Unitat d’Al·lèrgia, M. Ibero1, M. Castillo1, M. Vin Hospital de Terrassa, Terrassa, SPAIN, 2I + D, Laboratorios BIAL-Aristegui, Bilbao, SPAIN. CASE REPORT: A 16-year-old boy, diagnosed of rhinoconjunctivitis and sensitizated to: dust, molds, cat epithelium, dog epithelium and pollens (Olea europea, Cupressus arizonica, Lolium perenne, Cynodon dactylon, Parietaria judaica and Plantago lanceolata), that suffered from anaphylaxis after eating chicken with potatoes and artichoke. Several months later the patient suffered from swelling of the lips with almonds. Later he has tolerated chicken and potatoes. METHODS AND RESULTS: Skin prick tests were positive to extracts from: artichoke (crude and cooked) and nuts. Specific IgE (EAST) were: Flower of Artichoke (flower+head-food): 8.6 kU/l; Stem of Artichoke: 3.6 kU/l; Artichoke (head-food): 2.9 kU/l; peanut: 26.6 kU/l and almonds: 2.23 kU/l.. SDS-PAGE Immunoblotting with artichoke extract showed IgE-binding bands at 14 kDa, 15,5 kDa and 17 kDa. Cross reactivity was studied between the artichoke extract and the extract from these pollens to which the patient was sensitizated. EAST inhibition using artichoke extract as solid phase showed an inhibition of 60% with P. judaica pollen extract. Immunoblotting with artichoke extract showed a complete inhibition of the IgE-binding when patient serum was preincubated with P. judaica pollen extract. Inhibition studies showed the artichoke IgEbinding proteins cross-react with the lipid transfer protein (LTP) from peach. CONCLUSIONS: In our knowledge it is the first case of oral allergy to artichoke. The clinical evolution of the patient and the in vitro test suggest an initial sensitization by respiratory tract to pollen from P. judaica, and a later cross sensitization by digestive tract to artichoke.
187
Analysis of Ag-specific Ige Antibody, Total Serum Ige and Blood Eosinophil in Food Allergic Children J. Kim1, Y. Kim2, H. Lee2; 1Department of Pediatrics, Catholic University Medical College, Uijongbu-city, REPUBLIC OF KOREA, 2Department of Clinical Pathology, Catholic University Medical College, Uijongbu-city, REPUBLIC OF KOREA. RATIONALE: Challenge test remain standard for diagnosis of food allergy, they are difficult to perform and laboratory tests are usually used. We evaluated the correlations of food Ag-specific IgE class (MAST CLA, ImmunoCAP100), total IgE class (MAST CLA), total serum IgE, and blood eosinophil count in food allergic children. METHODS: A total of 293 children who visited allergy clinic were tested by Korean food panel of MAST CLA. RESULTS: In 17 of 38 children, who show higher than MAST CLA Class 2 without Ag-specific IgE class, were tested by ImmunoCAP100 for egg white, milk, wheat, soy bean, peanut, cat epithelium, dog epithelium, D. farinae, and D. pteronyssinus. There was statistical difference(p<0.05) between total IgE class0-1/0 and total IgE class 4 in the number of Agspecific IgE. Total IgE class 4 show larger number of Ag-specific IgE class than total IgE class 0-1/0. But no statistical differences between total IgE class0-1/0 and total IgE class 1, 2 and 3. Although no statistical differences between total serum IgE and the number of Ag-specific IgE class, there were statistical differences(p<0.05) between total IgE class 4 and total IgE class 0-1/0, 1, 2, and 3 respectively. No differences between blood eosinophil count and number of Ag-specific IgE class, total IgE class, total serum IgE level. 6(35.3%) of the 17 children, was higher than MAST CLA Class 2 without Ag-specific IgE class, show positive reaction in ImmunoCAP100 test. CONCLUSIONS: Total IgE class 4 show larger number of Ag-specific IgE class than that of total IgE class 0-1/0.
188
SATURDAY
Abstracts S47
J ALLERGY CLIN IMMUNOL VOLUME 117, NUMBER 2