title: Evaluation of different glycoforms of honeybee venom major allergen phospholipase A2 (Api m 1) produced in insect cells.
abstract: Allergic reactions to hymenoptera stings are one of the major reasons for IgE-mediated anaphylaxis. However, proper diagnosis using venom extracts is severely affected by molecular cross-reactivity. In this study recombinant honeybee venom major allergen phospholipase A2 (Api m 1) was produced for the first time in insect cells. Using baculovirus infection of different insect cell lines allergen versions providing a varying degree of cross-reactive carbohydrate determinants as well as a non glycosylated variant could be obtained as secreted soluble proteins in high yields. The resulting molecules were analyzed for their glycosylation and proved to show advantageous properties regarding cross-reactivity in sIgE-based assays. Additionally, in contrast to the enzymatically active native protein the inactivated allergen did not induce IgE-independent effector cell activation. Thus, insect cell-derived recombinant Api m 1 with defined CCD phenotypes might provide further insights into hymenoptera venom IgE reactivities and contribute to an improved diagnosis of hymenoptera venom allergy.
authors.affiliation: Institute of Biochemistry and Molecular Biology, University of Hamburg, Germany.
authors.forename: Simon
authors.initials: S
authors.lastname: Blank
authors.forename: Yvonne
authors.initials: Y
authors.lastname: Michel
authors.forename: Henning
authors.initials: H
authors.lastname: Seismann
authors.forename: Melanie
authors.initials: M
authors.lastname: Plum
authors.forename: Kerstin
authors.initials: K
authors.lastname: Greunke
authors.forename: Thomas
authors.initials: T
authors.lastname: Grunwald
authors.forename: Reinhard
authors.initials: R
authors.lastname: Bredehorst
authors.forename: Markus
authors.initials: M
authors.lastname: Ollert
authors.forename: Ingke
authors.initials: I
authors.lastname: Braren
authors.forename: Edzard
authors.initials: E
authors.lastname: Spillner
mesh_terms: D000485:Allergens / Q000737:chemistry* / Q000235:genetics / Q000276:immunology; D000818:Animals; D052179:Antigens, Plant / Q000737:chemistry* / Q000235:genetics / Q000276:immunology; D001514:Bee Venoms / Q000201:enzymology*; D001516:Bees / Q000201:enzymology*; D002460:Cell Line; D003429:Cross Reactions / Q000276:immunology; D006801:Humans; D006967:Hypersensitivity / Q000175:diagnosis; D007073:Immunoglobulin E / Q000097:blood / Q000276:immunology; D007299:Insect Bites and Stings / Q000175:diagnosis; D019476:Insect Proteins / Q000737:chemistry* / Q000235:genetics / Q000276:immunology; D016297:Mutagenesis, Site-Directed; D010741:Phospholipases A / Q000737:chemistry* / Q000235:genetics / Q000276:immunology; D051381:Rats; D011994:Recombinant Proteins / Q000737:chemistry* / Q000235:genetics / Q000276:immunology
chemical_list: D000485:Allergens; D052179:Antigens, Plant; D001514:Bee Venoms; D019476:Insect Proteins; D011994:Recombinant Proteins; D007073:Immunoglobulin E; D010741:Phospholipases A; C497838:Api m 1 allergen, Apis mellifera