Mutational analysis of the IgE-binding epitopes of P34/Gly m Bd 30K

Helm, Ricki M.; Cockrell, Gael; Connaughton, Cathie; West, Charles; Herman, Eliot M.; Sampson, Hugh A.; Bannon, Gary A.; Burks, A. Wesley

doi:10.1016/s0091-6749(00)90091-5

Cited by 90 publications

(57 citation statements)

References 23 publications

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“…However, structural studies cannot always be performed, because of limited amount of protein or unsuitable protein sizes for the method. Indeed to date, most allergen IgE epitopes have been located using overlapping synthetic peptides (8 -15 aa long) that cover the protein sequence, but such methods only identify linear stretches (34,35). N-terminal and C-terminal truncations (36 -38), fragmentation (39,40), and in-frame deletion (41) studies have also been successfully used to locate continuous B cell epitopes.…”

Section: Discussionmentioning

confidence: 99%

Construction of Hevein (Hev b 6.02) with Reduced Allergenicity for Immunotherapy of Latex Allergy by Comutation of Six Amino Acid Residues on the Conformational IgE Epitopes

Karisola

Mikkola

Kalkkinen

et al. 2004

The Journal of Immunology

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Recently we have established that IgE Abs bind to conformational epitopes in the N- and C-terminal regions of the major natural rubber latex allergen, hevein (Hev b 6.02). To identify the critical amino acid residues that interact with IgE, the hevein sequence was scanned by using site-specific mutations. Twenty-nine hevein mutants were designed and produced by a baculovirus expression system in insect cells and tested by IgE inhibition-ELISA using sera from 26 latex allergic patients. Six potential IgE-interacting residues of hevein (Arg5, Lys10, Glu29, Tyr30, His35, and Gln38) were identified and characterized further in detail. Based on these six residues, two triple mutants (HΔ3A, HΔ3B) and hevein mutant where all six residues were mutated (HΔ6), were designed, modeled, and produced. Structural and functional properties of these combinatory mutants were compared experimentally and in silico with those of recombinant hevein. The IgE-binding affinity of the mutants decreased by three to five orders of magnitude as compared with that of recombinant hevein. Skin prick test reactivity of the triple mutant HΔ3A was drastically reduced and that of the six-residue mutant HΔ6 was completely abolished in all patients examined in this study. The approach presented in this paper offers tools for identification and modification of amino acid residues on conformational epitopes of allergens that interact with IgE. Hevein with a highly reduced ability to bind IgE should provide a valuable candidate molecule for immunotherapy of latex allergy and is anticipated to have a low risk of systemic side effects.

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Section: Discussionmentioning

confidence: 99%

Construction of Hevein (Hev b 6.02) with Reduced Allergenicity for Immunotherapy of Latex Allergy by Comutation of Six Amino Acid Residues on the Conformational IgE Epitopes

Karisola

Mikkola

Kalkkinen

et al. 2004

The Journal of Immunology

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“…Subunits Gy1 and Gy5 are considered main epitopes for this protein (Schiller et al, 2014). Other soy proteins have been characterised and proposed as allergens, including the thiol-protease Gly m Bd 30k (Ogawa et al, 1991;Helm et al, 1998;Helm et al, 2000), and the Kunitz trypsin inhibitor (Moroz and Yang, 1980;Gu et al, 2001). …”

Section: Identified Allergensmentioning

confidence: 99%

Scientific Opinion on the evaluation of allergenic foods and food ingredients for labelling purposes

Products¹

2014

EFS2

115

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Following a request from the Food Safety Authority of Ireland, the EFSA Panel on Dietetic Products, Nutrition and Allergies (NDA Panel) was asked to deliver a scientific opinion on the evaluation of allergenic foods and food ingredients for labelling purposes. In view of the request, the NDA Panel decided to update its previous opinions relative to food ingredients or substances with known allergenic potential listed in Annex IIIa of 2003/89/EC, as amended. These include cereals containing gluten, milk and dairy products, eggs, nuts, peanuts, soy, fish, crustaceans, molluscs, celery, lupin, sesame, mustard and sulphites. The opinion relates to immunoglobulin (Ig)E-and non-IgE-mediated food allergy, to coeliac disease and to adverse reactions to sulphites in food, and it does not address non-immune-mediated adverse reactions to food. It includes information on the prevalence of food allergy in unselected populations, proteins identified as food allergens, cross-reactivities, the effects of food processing on the allergenicity of foods and ingredients, methods for the detection of allergens and allergenic foods, doses observed to trigger adverse reactions in sensitive individuals and risk assessment methodologies that have been used to derive individual and population thresholds for selected allergenic foods. The present opinion relates to immunoglobulin (Ig)E-and non-IgE-mediated food allergy, to coeliac disease and to adverse reactions to sulphites in food, and it does not address non-immune-mediated adverse reactions to food. For each food ingredient or substance listed in Annex IIIa, it includes information on the prevalence of food allergy in unselected populations, on proteins identified as food allergens, on cross-reactivities, on the effects of food processing on the allergenicity of foods and ingredients, on methods for the detection of allergens and allergenic foods, and on doses observed to trigger adverse reactions in sensitive individuals.Immune-mediated adverse reactions to foods manifest with clinical signs and symptoms of variable severity and duration, which may affect different organs and systems. Anaphylactic reactions to food are IgE mediated and may occur at any age. Non-IgE-mediated food allergy includes a wide range of diseases, including protein-induced enterocolitis and eosinophilic oesophagitis.A careful family and clinical history are the basis for diagnosis of food allergy. Food diaries, skin prick tests (SPTs), allergen-specific IgE measurements, food elimination diets and food challenges are part of the standard protocol for the diagnosis of food allergy. A positive SPT indicates sensitisation to the tested food, but it is not diagnostic of food allergy. Allergen-specific serum IgE antibodies similarly denote sensitisation to a particular food, but they are not diagnostic without a clinical history or food challenge. The use of atopy patch tests for the diagnosis of food allergy is controversial. Other available tests have no current role in the diagnosis of food allergy. Diag...

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“…The most frequently used strategy to identify amino acid positions critical for IgE Ab binding is to replace each amino acid with alanine (alanine scan) (38,39,(42)(43)(44). Instead of an alanine scan, amino acids located at homologous positions of nonallergenic tropomyosins were replaced in the allergenic peptides.…”

Section: Figurementioning

confidence: 99%

Reduced Allergenic Potency of VR9-1, a Mutant of the Major Shrimp Allergen Pen a 1 (Tropomyosin)

Reese

Viebranz

Leong-Kee

et al. 2005

The Journal of Immunology

106

101

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The major shrimp allergen, tropomyosin, is an excellent model allergen for studying the influence of mutations within the primary structure on the allergenic potency of an allergen; Pen a 1 allows systematic evaluation and comparison of Ab-binding epitopes, because amino acid sequences of both allergenic and nonallergenic tropomyosins are known. Individually recognized IgE Ab-binding epitopes, amino acid positions, and substitutions critical for IgE Ab binding were identified by combinatorial substitution analysis, and 12 positions deemed critical were mutated in the eight major epitopes. The mutant VR9-1 was characterized with regard to allergenic potency by mediator release assays using sera from shrimp-allergic subjects and sera from BALB/c, C57BL/6J, C3H/HeJ, and CBA/J mice sensitized with shrimp extract using alum, cholera toxin, and Bordetella pertussis, as adjuvants. The secondary structure of VR9-1 was not altered; however, the allergenic potency was reduced by 90–98% measuring allergen-specific mediator release from humanized rat basophilic leukemia (RBL) cells, RBL 30/25. Reduced mediator release of RBL-2H3 cells sensitized with sera from mice that were immunized with shrimp extract indicated that mice produced IgE Abs to Pen a 1 and to the same epitopes as humans did. In conclusion, data obtained by mapping sequential epitopes were used to generate a Pen a 1 mutant with significantly reduced allergenic potency. Epitopes that are relevant for human IgE Ab binding are also major binding sites for murine IgE Abs. These results indicate that the murine model might be used to optimize the Pen a 1 mutant for future therapeutic use.

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Mutational analysis of the IgE-binding epitopes of P34/Gly m Bd 30K

Cited by 90 publications

References 23 publications

Construction of Hevein (Hev b 6.02) with Reduced Allergenicity for Immunotherapy of Latex Allergy by Comutation of Six Amino Acid Residues on the Conformational IgE Epitopes

Construction of Hevein (Hev b 6.02) with Reduced Allergenicity for Immunotherapy of Latex Allergy by Comutation of Six Amino Acid Residues on the Conformational IgE Epitopes

Scientific Opinion on the evaluation of allergenic foods and food ingredients for labelling purposes

Reduced Allergenic Potency of VR9-1, a Mutant of the Major Shrimp Allergen Pen a 1 (Tropomyosin)

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