Eberlein-Konig, B. et al. Comparison of basophil activation tests using CD63 or CD203c expression in patients with insect venom allergy. Allergy 61, 1084-1085

Department of Dermatology and Allergy Biederstein, Technical University Munich, Munich, Germany.
Allergy (Impact Factor: 6.03). 10/2006; 61(9):1084-5. DOI: 10.1111/j.1398-9995.2006.01122.x
Source: PubMed


Flow cytometric basophil activation tests have been developed as cellular tests for in vitro diagnosis of IgE-mediated reactions. Different activation markers (CD63 or CD203c) with distinct ways of regulation have been used after stimulation with various allergens.
It was the aim of the present study to compare basophil activation tests by measuring both CD63 and CD203c upregulation in patients with insect venom allergy.
43 patients with a history of insect venom anaphylaxis were examined. A careful allergy history was taken, and skin tests and determination of specific IgE-antibodies were performed. Basophil activation tests (BAT) using CD63 or CD203c expression were done after stimulation with different concentrations of bee and wasp venom extracts. 25 healthy subjects with negative history of insect venom allergy were studied as controls.
The CD203c protocol showed a slightly higher sensitivity than the CD63 protocol (97% vs. 89%) with regard to patients' history. The magnitude of basophil response was higher with CD203c in comparison to CD63 for both insect venoms. Specificity was 100% for the CD63 protocol and 89% for the CD203c protocol with regard to controls with negative history and negative RAST.
These results support the reliability of basophil activation tests using either CD63 or CD203c as cellular tests in the in vitro diagnosis of patients with bee or wasp venom allergy with a slightly higher sensitivity for the CD203c protocol.

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    • "To diagnose specific-venom allergy, investigations to demonstrate the presence of sIgE are used to confirm the causative species and thus select an appropriate venom extract for VIT [10]. Methods include intradermal venom skin testing, quantitation of sIgE in serum and functional assays such as the histamine release test and flow-cytometry analysis of CD63 and CD203c upregulation on basophils [11], [12], [13], [14]. In vitro laboratory methods are thought to have lower sensitivity compared to intradermal venom skin testing [10]. "
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    ABSTRACT: We adapted DELFIA™ (dissociation-enhanced lanthanide fluoroimmunoassay), a time resolved fluorescence method, to quantitate whole venom specific and allergenic peptide-specific IgE (sIgE), sIgG(1) and sIgG(4) in serum from people clinically allergic to Australian native ant venoms, of which the predominant cause of allergy is jack jumper ant venom (JJAV). Intra-assay CV was 6.3% and inter-assay CV was 13.7% for JJAV sIgE. DELFIA and Phadia CAP JJAV sIgE results correlated well and had similar sensitivity and specificity for the detection of JJAV sIgE against intradermal skin testing as the gold standard. DELFIA was easily adapted for detecting sIgE to a panel of other native ant venoms.
    Full-text · Article · Jan 2011 · PLoS ONE
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    • "The ROC curves for CD63 and CD203c and the threshold for positivity were almost identical for CD63 and CD203c, suggesting that the two markers measure similar aspects of basophil activation. Although Boumiza has disputed this [26], it has since been confirmed by a number of clinically oriented publications [24,27,28]. We have previously shown that choice of fluorochome and lysis procedure had favoured CD203c over CD63, illustrating the need to consider all aspects of an experiment [6]. "
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    ABSTRACT: Subcutaneous Immunotherapy (SCIT) modifies the allergic response and relieves allergic symptoms. SCIT is the only and a very effective treatment for insect venom allergy. We hypothesized that basophil sensitivity, measured through the basophil activation test, would decrease during SCIT up dosing. Expression of CD203c was compared to CD63 as marker for basophil activation, using a Bland Altman plot and ROC curves. Patients (n = 18) starting subcutaneous SCIT for wasp allergy with an up dosing scheme of 7 to 11 weeks were enrolled. Heparinised blood samples were drawn at weeks 1-4, 7 and at the first maintenance visit. Basophils were stimulated at 7 log dilutions of V. vespula allergen for 15 min, and were stained with CD203c and CD63. Basophils were identified as CD203c+ leukocytes, and the proportion of CD63+ and CD203c+ cells were plotted against allergen concentration. A sigmoid curve was fitted to the points, and the allergen concentration at which half of the maximal activation was achieved, LC50, was calculated. In another series of experiments, LC50 calculated in whole blood (AP) was subtracted from LC50 calculated with basophils suspended in plasma from a nonatopic donor (HS) to determine the protective effect of soluble factors in blood of patients treated with SCIT. Heparin blood basophil activation was similar through CD63 and CD203c. Basophils were significantly more sensitized three weeks after initiation of SCIT compared to baseline (p < 0,01). The difference in LC50 increased by 1,04 LC50 units (p = 0,04) in patients that had just achieved maintenance dose compared with patients before initiating SCIT. When maintenance allergen concentrations had been reached, an increase in the protective plasma component was documented. Blood basophil concentration was marginally reduced by SCIT. Basophil activation is a versatile and sensitive tool that measures changes in the humoral immune response to allergen during SCIT.
    Full-text · Article · Feb 2010 · Clinical and Molecular Allergy
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    ABSTRACT: Introduction: 'Gold standard' in the diagnosis of atopic disease are skin prick tests and specific IgE evaluation. Well- -established in vitro tests, such as the histamine release test, the leukotriens release test and the flow cytometric basophil activation test can be very helpful in diagnostics, especially when the skin prick test is contraindicated. The aim of this study was to evaluate the usefulness of antigen CD203c expression, as a marker of basophil activation by grass pollen or D. pteronyssinus antigens. Material and methods: Peripheral blood from 13 allergic patients and nine healthy volunteers was analysed. Basophils activation was measured by the breakdown of antigen CD203c expression with Allergenicity Kit (Beckman Coulter), using Cytomics FC 500 flow cytometer (Beckman Coulter). Results: The sensitivity was 92,3% and specificity of test was 100%. 50.95 ± 15.7% of basophils (median 49.7%, 1.91- -72.42%) were activated after grass pollen stimulation in atopic patients sensitised to this allergen, in comparison to 1.91% (0.00-7.96%) in control patients (p = 0.002). The percentage of activated basophils after D. pteronyssinus antigens stimulation was 40.6 ± 25.2% in allergic patients, compared to only 2.51 ± 1 96% of basophils from non-atopic controls (p = 0.0003). Basophils from 21 patients responded after anti-IgE stimulation (44.1 ± 18.9%), and none of the analysed samples was activated after PBS stimulation (2.03 ± 1.19%, p < 0.0001). Conclusions: These results demonstrate that basophil activation test based on antigen CD203c expression is very accurate in the diagnosis of atopic diseases.
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