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Hymenoptera venom allergy is known to cause life-threatening and sometimes fatal IgE-mediated anaphylactic reactions in allergic individuals. About 30-50% of patients with insect venom allergy have IgE antibodies that react with both honeybee and yellow jacket venom. Apart from true double sensitisation, IgE against cross-reactive carbohydrate determinants (CCD) are the most frequent cause of multiple reactivities severely hampering the diagnosis and design of therapeutic strategies by clinically irrelevant test results. In this study we addressed allergenic cross-reactivity using a recombinant approach by employing cell lines with variant capacities of alpha-1,3-core fucosylation. The venom hyaluronidases, supposed major allergens implicated in cross-reactivity phenomena, from honeybee (Api m 2) and yellow jacket (Ves v 2a and its putative isoform Ves v 2b) as well as the human alpha-2HS-glycoprotein as control, were produced in different insect cell lines. In stark contrast to production in Trichoplusia ni (HighFive) cells, alpha-1,3-core fucosylation was absent or immunologically negligible after production in Spodoptera frugiperda (Sf9) cells. Consistently, co-expression of honeybee alpha-1,3-fucosyltransferase in Sf9 cells resulted in the reconstitution of CCD reactivity. Re-evaluation of differentially fucosylated hyaluronidases by screening of individual venom-sensitised sera emphasised the allergenic relevance of Api m 2 beyond its carbohydrate epitopes. In contrast, the vespid hyaluronidases, for which a predominance of Ves v 2b could be shown, exhibited pronounced and primary carbohydrate reactivity rendering their relevance in the context of allergy questionable. These findings show that the use of recombinant molecules devoid of CCDs represents a novel strategy with major implications for diagnostic and therapeutic approaches.

Original publication

DOI

10.1016/j.molimm.2009.10.005

Type

Journal article

Journal

Molecular immunology

Publication Date

01/2010

Volume

47

Pages

799 - 808

Addresses

Institute of Biochemistry and Molecular Biology, University of Hamburg, Hamburg, Germany.

Keywords

Cell Line, Subcellular Fractions, Animals, Humans, Hypersensitivity, Hyaluronoglucosaminidase, Fucosyltransferases, Carbohydrates, Fucose, Blood Proteins, Immunoglobulin E, Recombinant Proteins, Bee Venoms, Wasp Venoms, Immunoblotting, Enzyme-Linked Immunosorbent Assay, Immunization, Electrophoresis, Polyacrylamide Gel, Cloning, Molecular, Cross Reactions, Phenotype, alpha-2-HS-Glycoprotein