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.

6533b7dcfe1ef96bd1273563

RESEARCH PRODUCT

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.

Olli H. Laitinen Markku S. Kulomaa Kristiina Turjanmaa Olli T. Pentikäinen Harri Alenius Kari J. Airenne Jari Mikkola Nisse Kalkkinen Susanna Repo Timo Reunala Timo Palosuo Mark S. Johnson Piia Karisola

subject

Male Models Molecular Protein Conformation Mutant Immunoglobulin E medicine.disease_cause Epitope law.invention Epitopes 0302 clinical medicine Protein structure Allergen law Immunology and Allergy Combinatorial Chemistry Techniques Child 0303 health sciences biology Chemistry Middle Aged Recombinant Proteins 3. Good health Biochemistry Latex allergy Recombinant DNA Female Plant Lectins Protein Binding Adult Adolescent Immunology Mutagenesis (molecular biology technique)Binding Competitive 03 medical and health sciences Latex Hypersensitivity medicine Humans Point Mutation 030304 developmental biology Aged Allergens Immunoglobulin E medicine.disease 030228 respiratory system Amino Acid Substitution Desensitization Immunologic biology.protein Mutagenesis Site-Directed Binding Sites Antibody Antimicrobial Cationic Peptides

description

Abstract 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.

year	journal	country	edition	language
2004-02-07	Journal of immunology (Baltimore, Md. : 1950)

10.4049/jimmunol.172.4.2621 https://pubmed.ncbi.nlm.nih.gov/14764736