rApi m 3 and rApi m 10 improve detection of honey bee sensitization in Hymenoptera venom-allergic patients with double sensitization to honey bee and yellow jacket venom.

Recombinant allergens improve the diagnostic precision in Hymenoptera venom allergy (HVA), in particular in patients with double sensitization to both honey bee (HBV) and yellow jacket venom (YJV). While currently available vespid allergens allow the detection of >95% of YJV-allergic patients, the sensitization frequency to the only available HBV marker allergen rApi m 1 in HBV-allergic patients is lower. Here, we demonstrate that sIgE to additional HBV marker allergens rApi m 3 and rApi m 10 allows the detection of genuine HBV sensitization in 46-65% of Api m 1 negative sera. This is of particular relevance in patients with double sensitization to HBV and YJV that did not identify the culprit insect. Addition of sIgE to rApi m 3 and rApi m 10 provides evidence of HBV sensitization in a large proportion of rApi m 1-negative patients and thus provides a diagnostic marker and rationale for VIT treatment with HBV, which otherwise would have been missing.

The major royal jelly proteins 8 and 9 (Api m 11) are glycosylated components of Apis mellifera venom with allergenic potential beyond carbohydrate-based reactivity.

BACKGROUND: As hymenoptera venoms are one of the allergen sources causing the highest incidence of anaphylaxis and sometimes fatal consequences, the detailed characterization of all venom allergens is imperative for design of component-resolved diagnostic approaches and improved intervention strategies. OBJECTIVE: Our aim was the immunochemical characterization of major royal jelly proteins (MRJP) 8 and 9, both components identified in honeybee venom (HBV) and putative allergens. METHODS: Both MRJPs were recombinantly produced as soluble differentially glycosylated proteins providing a defined degree of reactivity to cross-reactive carbohydrate determinants (CCD) in insect cells. Allergen-specific IgE(sIgE) reactivity of HBV-allergic patients was analysed by ELISA and immunoblotting. RESULTS: MRJP8 and MRJP9 were identified as venom components by MS-based proteomic analyses. In a population of 47 HBV-allergic patients, reactivities with CCD-carrying MRJPs were in the range of 56% (61%), underlining the contribution of CCDs to allergen-binding. Beyond CCD-reactivity, 15% of patients showed sIgE reactivity with MRJP8 and 34% with MRJP9 respectively. These reactivities roughly in the range of Api m 2 render the MRJPs minor, but important allergens. CONCLUSION AND CLINICAL RELEVANCE: The glycosylated MRJP8 and MRJP9 of HBV have IgE-sensitizing potential in HBV-allergic patients beyond CCD reactivity and have to be considered as allergens, which might be potentially important for a fraction of venom allergic patients. They are valuable tools to elucidate individual component-resolved reactivity profiles of venom allergic patients and to provide insights into the role of particular venom components. Due to their allergenic properties, MRJP8 and MRJP9 were designated as isoallergens Api m 11.0101 and Api m 11.0201 respectively.