Basophil Activation Test Using Recombinant Allergens: Highly Specific Diagnostic Method Complementing Routine Tests in Wasp Venom Allergy (original) (raw)
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In vitro basophil activation using CD63 expression in patients with bee and wasp venom allergy
Journal of investigational allergology & clinical immunology, 2006
The diagnosis of insect venom allergy and the indication for specific immunotherapy is based on history, skin tests and demonstration of hymenoptera venom-specific IgE-antibodies. Cellular tests can add useful information but the role of basophil activation tests for the different venoms has to be elucidated further. We evaluated positive reactions in a basophil activation test using CD63 expression as marker independently for bee or wasp venom in patients with hymenoptera allergy. Fifty-seven patients with a history of insect venom anaphylaxis were examined (12 x bee venom, 39 x wasp venom, 6 x bee plus wasp venom). Skin tests and determination of specific IgE-antibodies were performed. Basophil activation test (BAT) using CD63 expression was performed after stimulation with different concentrations of bee and wasp venom. The BAT is based on double staining with anti-IgE antibodies and anti-CD63 and subsequent determination of the percentage of activated basophils by flow cytometry...
Journal of Allergy and Clinical Immunology, 2012
Background: Specific IgE (sIgE) antibodies to both bee and wasp venom can be due to a sensitivity to both insect venoms or due to cross-reactive carbohydrate determinants (CCDs). Objective: Investigating whether a basophil activation test (BAT) with both venoms as well as with bromelain and horseradish peroxidase (HRP) or recombinant allergen-based IgE testing can improve the diagnostic procedure. Methods: Twenty-two Hymenoptera-venom allergic patients with sIgE antibodies to both bee and wasp venom were studied. sIgE antibodies to MUXF3 CCD, bromelain, HRP, rApi m 1, and rVes v 5 were determined, and a BAT (Flow2 CAST) with venom extracts, bromelain, and HRP was performed. Further recombinant allergen-based IgE testing was done by using an ELISA, if required. The reactivity of basophils was calculated from the insect venom concentration at half-maximum stimulation. Results: Double positivity/double negativity/single positivity to rApi m 1 and rVes v 5 was seen in 12/1/9 patients. Further recombinant allergen-based IgE testing in the last ones revealed positive results to the other venom in all cases except one. BAT was double positive/double negative/single positive in 6/2/14 patients. Four patients with negative results in sIgE antibodies to CCDs had positive results in BAT. BAT with bromelain/HRP showed a sensitivity of 50%/81% and a specificity of 91%/90%. Conclusion: Component-resolved IgE testing elucidates the pattern of double positivity, showing a majority of true double sensitizations independent of CCD sensitization. BAT seems to add more information about the culprit insect even if the true clinical relevance of BAT is not completely determined because of ethical limitations on diagnostic sting challenges. BAT with HRP is a good method to determine sensitivity to CCDs.
The CD63 basophil activation test in Hymenoptera venom allergy: a prospective study
Allergy, 2004
The basophil activation test (BAT), which relies on flow cytometric quantitation of the allergen-induced up-regulation of the granule-associated marker CD63 in peripheral blood basophils, has been suggested to be a useful approach in detecting responsiveness to allergens. The purpose of this study was to establish the usefulness of the BAT with regard to the clinical history and current diagnostic tools in Hymenoptera venom allergy using a prospective study design. Fifty-seven consecutive patients allergic to Hymenoptera venom as defined by a systemic reaction after an insect sting, and 30 age- and sex-matched control subjects with a negative history were included. The degree and nature of sensitization was confirmed by skin testing, specific immunoglobulin E (IgE), serum tryptase levels and BAT. In the nonallergic control group only analysis of specific IgE and BAT were performed. Correlation of BAT, skin test and specific IgE, respectively, with the clinical history in the allergic group was termed as sensitivity and in the control group as specificity. Twenty one of 23 (91.3%) bee venom allergic patients and 29 of 34 (85.3%) patients allergic to wasp and hornet venom tested positive in BAT. The overall sensitivity of BAT, specific IgE and skin tests were 87.7, 91.2 and 93.0%, respectively. The overall specificities were 86.7% for BAT and 66.7% for specific IgE. No correlation between the severity of clinical symptoms and the magnitude of basophil activation was observed. The BAT seems to be an appropriate method to identify patients allergic to bee or wasp venom with a comparable sensitivity to standard diagnostic regimens. The higher specificity of BAT as compared with specific IgE makes this test a useful tool in the diagnosis of Hymenoptera venom allergy.
Basophil activation test in the diagnostics of hymenoptera venom allergy
Bratislava Medical Journal, 2021
The aim of the study was to analyse the diagnostic performance of the basophil activation test (BAT), to compare the diagnostic reliability of BATs performed with different types of allergens, which are available in Slovakia and to verify the correlation between the symptom severity and the sensitivity and specifi city of the BAT in 114 patients with suspected hymenoptera venom allergy (HVA). BACKGROUND: Diagnosis of the HVA and the identifi cation of the appropriate venom for immunotherapy are in Slovakia based on detailed patient'medical history, skin tests and detection of sIgE. In unclear cases, where the clinical decision regarding the relevant insect species for immunotherapy is diffi cult, the cellular tests are recommended in several countries, such as Sweden, Spain, Germany, Denmark and Italy. In Slovakia, BAT is not adapted as s part of routine diagnostic work-up. METHODS: The identifi cation of the culprit hymenoptera species among 114 patients was based on detailed history, skin tests and detection of sIgE. Obtained results were compared with the results acquired by the BAT. RESULTS: The sensitivity of the BAT was 80.8 % and the specifi city was 87.8 %. The sensitivity of the BAT was higher when using Soluprick SQ Allergens, but the specifi city was higher with BŰHLMANN CAST Allergens. In the study no correlation between the symptom severity and the sensitivity and specifi city of the BAT was observed. CONCLUSIONS: The results show that the BAT can be recommended in the identifi cation of the appropriate venom for immunotherapy, the only specifi c treatment that is currently available for patients with HVA. Allergen source is one of critical factors in diagnostic reliability of the BAT (Tab. 4, Ref.
High sensitivity of basophils predicts side-effects in venom immunotherapy
Allergy, 2005
Venom immunotherapy (VIT) is an efficient and only viable option for prophylaxis in patients allergic to Hymenoptera venom (1). However, local and systemic side-effects of this treatment, especially during incremental phase of VIT, are not rare (2). Risk and pathogenic factors for those adverse reactions are so far poorly understood. VIT with honeybee venom induces sideeffects more often than treatment with vespid venoms (3-6), which is further affected by different treatment protocols (7-9). The frequency of local and mild systemic reactions could be reduced by pretreatment with antihistamines (10), yet this is not obvious for the occurrence of severe anaphylactic reactions in patient treated with bee venom (11). Recently, the association between elevated serum tryptase level with or without mastocytosis and the severity of anaphylactic Hymenoptera sting reactions was shown (12, 13), but there is no evidence that basal serum tryptase level correlates to the frequency or severity of side-effects to VIT. At the moment there is no reliable test, which can predict higher risk for adverse reactions during VIT. Flow cytometry quantification of basophil activation by measurement of CD63 expression (14) is a very sensitive and specific cellular in vitro method for identification of Hymenoptera venom allergy (13, 15, 16). As basophils play an important role in the pathogenesis of allergic anaphylactic reactions (17), we examined the hypothesis whether basophil responsiveness might be connected with side reactions to VIT. Therefore, the aim of our study was to evaluate, by measuring CD63 surface expression as a marker of basophil activation, whether there is any correlation between basophil in vitro sensitivity to allergen-specific stimulation and the occurrence of VIT-induced side-effects. Materials and methods Study subjects Thirty-four patients undergoing VIT (median age of 41 years; age range: 17-70; 15 women) with a history of at least one systemic anaphylactic reaction of Mueller grades II-IV (18) after Background: Systemic side-effects of venom immunotherapy (VIT) represent a considerable problem in the treatment of patients allergic to Hymenoptera venom. We examined the hypothesis whether basophil responsiveness might be connected with the adverse reactions to VIT. Methods: Basophil surface expression of activation marker CD63 induced by different concentrations of honeybee and wasp venom (0.1 and 1 lg/ml) was measured by flow cytometry in 34 patients with history of systemic anaphylactic reactions to Hymenoptera sting just before rush honeybee or wasp VIT. Results: Eleven of 34 patients had systemic anaphylactic reaction (Mueller grades I-III) and one patient a large local reaction to VIT. In those 12 patients, median percentage of activated basophils after stimulation with VIT-specific venom in concentration of 0.1 lg/ml was 99% (range: 17-195) of value reached with stimulation with 1 lg/ml. Side-effects occurred in all patients with 0.1/1 ratios over 92% (eight of 12). In contrast, in 22 patients with no side-effects, the median 0.1/1 ratio was 25% (range: 2-92). These concentration-dependent activation ratios were significantly different between the groups with and without side reactions (P < 0.0001). We also show significant positive correlation of the occurrence/clinical grade of the side-effects with individual ratios of CD63 basophil response (r ¼ 0.73, P < 0.0001). Conclusion: The results suggest that increased basophil sensitivity to allergenspecific in vitro stimulation is significantly associated with major side-effects of VIT.
Clinical and Molecular Allergy
Background Allergy to Apis dorsata (Giant Asian Honeybee) venom is the commonest insect allergy in Sri Lanka and South East Asia. However, laboratory diagnosis is difficult as the pure venom and diagnostic reagents are not commercially available. Objective This study assessed the use of four recombinant allergens of A. mellifera venom and the passive basophil activation test in the diagnosis of A. dorsata venom anaphylaxis. Methods Serum IgE levels to four recombinant allergens of A. mellifera, rApi m 1, 2, 5 and 10 were assessed and compared with serum IgE to the crude venom of A. mellifera or V. vulgaris by Phadia ImmunoCAP, in patients who developed anaphylaxis to A. dorsata stings. Basophil activation in response to venom of A. dorsata or V. affinis was assessed using a passive basophil activation test. Association of the severity of the reaction with basophil activation was compared. Results rApi m 1 and 10 combinedly had significant correlation (r = 0.722; p < 0.001) with t...
International Archives of Allergy and Immunology, 2013
Background: Previous reports suggest the usefulness of basophil activation testing (BAT) in Hymenoptera-allergic patients with negative venom-specific IgE antibodies. We sought to evaluate the diagnostic utility of this testing in a routine clinical laboratory setting. Materials and Methods: Twenty-one patients with anaphylactic reactions to Hymenoptera sting (median grade III) and negative venom-specific IgE were routinely and prospectively tested with BAT. Results: We were able to diagnose 81% (17 of 21) of patients with BAT and 57% (12 of 21) with intradermal skin testing. Three wasp venom-allergic patients showed IgE positivity to rVes v 5. Four patients (19%) were negative for all tests. In the case of double-positive BAT, the culprit insect correlated with the venom that induced a significantly higher basophil response. Conclusions: BAT allows the identification of severe Hymenoptera-allergic patients with negative specific IgE and skin tests. The routine use of this cellular test should facilitate prescription of venom immunotherapy in complex cases with inconclusive diagnostic results.