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Involvement of carbohydrate epitopes in the IgE response of celery-allergic patients.
Int Arch Allergy Immunol 1999; 120(1):30-42IA

Abstract

BACKGROUND

This study was performed to get further insights into antibody responses to cross-reactive carbohydrate determinants (CCD), including initial experiments to prove the biological activity of anti-CCD IgE. Earlier studies have shown that IgE specific for CCD occurs in about 25% of celery-allergic patients. The clinical significance of these antibody specificities is doubtful.

METHODS

Patient sera were selected on the basis of a positive case history of celery allergy and multiple binding to high molecular weight celery allergens on immunoblots. Specific IgE to native and heated celery tuber was determined by the enzyme allergosorbent test (EAST). N-glycans were purified after extensive digestion of specific glycoproteins, such as pineapple stem bromelain, bovine fibrin, and human IgG, and used as antigens in an IgE ELISA as well as in EAST and immunoblotting inhibition experiments. Dose-related histamine release was performed with BSA neoglycoproteins containing 3-4 units of the purified glycopeptides.

RESULTS

Seven celery-allergic patients were identified who clearly presented IgE against the N-glycan purified from bromelain which is a common structure within the plant kingdom. Chemical defucosylation showed that alpha1, 3-fucose is a key structure for IgE binding. In patients with anti-CCD IgE, the maximal inhibition of celery EAST by the bromelain glycan ranged from 22 to 100%. Inhibition of celery immunoblots by preincubation of patient serum with this glycan led to a quenching of multiple bands at masses >40 kD. After linking the bromelain glycopeptide to BSA, a strong dose-related histamine release was obtained in a celery-allergic patient, occurring at lower concentrations than with the recombinant major protein allergen from celery, Api g 1.

CONCLUSIONS

Our results demonstrate that IgE specific for CCD is common in celery-allergic patients, and can represent the major proportion of IgE against this food. alpha1, 3-fucose was confirmed to be an essential part of the IgE epitope. Immunoblotting inhibition indicated the presence of this carbohydrate determinant on multiple glycoproteins in celery extract. Although histamine release was only performed in 1 patient, our data show that proteins carrying multiple glycan units can be biologically active in patients sensitized to CCD.

Authors+Show Affiliations

Department of Allergology, Paul-Ehrlich-Institute, Langen, Germany.No affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article

Language

eng

PubMed ID

10529586

Citation

Fötisch, K, et al. "Involvement of Carbohydrate Epitopes in the IgE Response of Celery-allergic Patients." International Archives of Allergy and Immunology, vol. 120, no. 1, 1999, pp. 30-42.
Fötisch K, Altmann F, Haustein D, et al. Involvement of carbohydrate epitopes in the IgE response of celery-allergic patients. Int Arch Allergy Immunol. 1999;120(1):30-42.
Fötisch, K., Altmann, F., Haustein, D., & Vieths, S. (1999). Involvement of carbohydrate epitopes in the IgE response of celery-allergic patients. International Archives of Allergy and Immunology, 120(1), pp. 30-42.
Fötisch K, et al. Involvement of Carbohydrate Epitopes in the IgE Response of Celery-allergic Patients. Int Arch Allergy Immunol. 1999;120(1):30-42. PubMed PMID: 10529586.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Involvement of carbohydrate epitopes in the IgE response of celery-allergic patients. AU - Fötisch,K, AU - Altmann,F, AU - Haustein,D, AU - Vieths,S, PY - 1999/10/26/pubmed PY - 1999/10/26/medline PY - 1999/10/26/entrez SP - 30 EP - 42 JF - International archives of allergy and immunology JO - Int. Arch. Allergy Immunol. VL - 120 IS - 1 N2 - BACKGROUND: This study was performed to get further insights into antibody responses to cross-reactive carbohydrate determinants (CCD), including initial experiments to prove the biological activity of anti-CCD IgE. Earlier studies have shown that IgE specific for CCD occurs in about 25% of celery-allergic patients. The clinical significance of these antibody specificities is doubtful. METHODS: Patient sera were selected on the basis of a positive case history of celery allergy and multiple binding to high molecular weight celery allergens on immunoblots. Specific IgE to native and heated celery tuber was determined by the enzyme allergosorbent test (EAST). N-glycans were purified after extensive digestion of specific glycoproteins, such as pineapple stem bromelain, bovine fibrin, and human IgG, and used as antigens in an IgE ELISA as well as in EAST and immunoblotting inhibition experiments. Dose-related histamine release was performed with BSA neoglycoproteins containing 3-4 units of the purified glycopeptides. RESULTS: Seven celery-allergic patients were identified who clearly presented IgE against the N-glycan purified from bromelain which is a common structure within the plant kingdom. Chemical defucosylation showed that alpha1, 3-fucose is a key structure for IgE binding. In patients with anti-CCD IgE, the maximal inhibition of celery EAST by the bromelain glycan ranged from 22 to 100%. Inhibition of celery immunoblots by preincubation of patient serum with this glycan led to a quenching of multiple bands at masses >40 kD. After linking the bromelain glycopeptide to BSA, a strong dose-related histamine release was obtained in a celery-allergic patient, occurring at lower concentrations than with the recombinant major protein allergen from celery, Api g 1. CONCLUSIONS: Our results demonstrate that IgE specific for CCD is common in celery-allergic patients, and can represent the major proportion of IgE against this food. alpha1, 3-fucose was confirmed to be an essential part of the IgE epitope. Immunoblotting inhibition indicated the presence of this carbohydrate determinant on multiple glycoproteins in celery extract. Although histamine release was only performed in 1 patient, our data show that proteins carrying multiple glycan units can be biologically active in patients sensitized to CCD. SN - 1018-2438 UR - https://www.unboundmedicine.com/medline/citation/10529586/Involvement_of_carbohydrate_epitopes_in_the_IgE_response_of_celery_allergic_patients_ L2 - https://www.karger.com?DOI=10.1159/000024217 DB - PRIME DP - Unbound Medicine ER -