Basophil activation test based on the expression of CD203c in the diagnostics of cow milk allergy in children.
ABSTRACT Detection of allergen-induced basophil activation by flow cytometry has been shown to be a useful tool for allergy diagnosis.
The aim of this study was to assess the potential of this technique for the diagnosis of pediatric cow milk allergy.
The quantification of total and specific IgE and basophil activation test were performed to evaluate cow milk allergic (n = 9), and non-allergic children (n = 15).
Allergen-induced basophil activation was detected as a CD203c up-regulation. The expression of CD203c antigen on basophils was measured with flow cytometry. The antigen CD203c was detected on 15.4 ±10.2% basophils from allergic children after incubation with specific allergens in concentration 1:10, whereas in the control group there were 3.0 ±1.5% of basophils positive for this molecule (P<0.05). Stimulation with allergen diluted 1:500 resulted in activation of 15.3 ±11.2% of basophils in allergic children and 3.8 ±2.3% of cells in the control group (P<0.05). Positive results of an allergenicity test (above the cut- off level of 10%) were obtained in 7 out of the 9 allergic children. In 5 cases, the cut-off level was reached with both dilution of allergens (1:10 and 1:500). In 1 patient, positive stimulation was observed after stimulation with allergen diluted 1:10 and in another case only 1:500 resulted in stimulation of more then 10% of basophils. In no child of the control group, stimulation above 10% was noted.
This study demonstrates that the analysis of allergen-induced CD203c up-regulation with flow cytometry is a reliable tool for the diagnosis of cow milk allergy in pediatric patients, with sensitivity similar to routine diagnostic tests and a higher specificity.
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Article: Basophil activation test based on the expression of CD203c in the diagnostics of cow milk allergy in children.
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- "Another test method includes the measurement of allergen-induced basophil activation by flow cytometry by analysis of the basophil activation markers CD203c or CD63. These basophil tests help to determine the clinical course of CMA and to make the decision if food challenges should be performed [21,171–175]. "
ABSTRACT: The first adverse reactions to cow's milk were already described 2000 years ago. However, it was only 50 years ago that several groups started with the analysis of cow's milk allergens. Meanwhile the spectrum of allergy eliciting proteins within cow's milk is identified and several cow's milk allergens have been characterized regarding their biochemical properties, folding and IgE binding epitopes. The diagnosis of cow's milk allergy is diverse ranging from fast and cheap in vitro assays to elaborate in vivo assays. Considerable effort was spent to improve the diagnosis from an extract-based into a component resolved concept. There is still no suitable therapy available against cow's milk allergy except avoidance. Therefore research needs to focus on the development of suitable and safe immunotherapies that do not elicit severe side effect.Methods 08/2013; 66(1). DOI:10.1016/j.ymeth.2013.08.005 · 3.65 Impact Factor
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ABSTRACT: A wide range of reported evidence in the literature has shown that the quantification of basophil activation by flow cytometry (basophil activation test, BAT) has proven to be a useful tool for the assessment of immediate-type responses to allergens mediated by IgE or other mechanisms in allergic patients. The usefulness of BAT in anaphylactic adverse reactions, late-onset allergy and immunotherapy follow-up has also been demonstrated. To date, most BAT studies reported in the literature involved the capture of basophils only with a fluorochrome-labeled anti-IgE antibody and application of CD63 upregulation in order to evaluate the basophil response to allergens. Many issues need to be addressed, such as optimizing the analytical performance of the test, checking preanalytical conditions, the selection of the flow cytometry best gating protocol, the introduction of new algorithms and parameters, the search for new activation markers and the introduction of anti-IgE controls. BAT is certainly a useful technique, also for isolated cases of hypersensitivity to various other compounds and drugs, and an update of its application is certainly an interesting topic to expand the debate on allergy diagnosis.International Archives of Allergy and Immunology 01/2012; 158(2):99-114. DOI:10.1159/000331312 · 2.67 Impact Factor
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ABSTRACT: INTRODUCTION: Fifteen to sixty percent of cystic fibrosis patients harbor Aspergillus fumigatus (Af) in their airways (CF-AC) and some will develop allergic bronchopulmonary aspergillosis (CF-ABPA). Since basophils play a key role in allergy, we hypothesized that they would display alterations in CF-ABPA patients compared to CF-AC or patients without Af colonization (CF). METHODS: Using flow cytometry, we measured CD203c, CD63 and CD123 levels on basophils from CF-ABPA (N=11), CF-AC (N=14), and CF (N=12) patients before and after ex vivo stimulation with Af allergens. RESULTS: Baseline CD203c was increased in basophils from CF-ABPA compared to CF-AC and CF patients. Af extract and recombinant Aspf1 stimulated basophils from CF-ABPA patients to markedly upregulate CD203c, along with modest upregulation of CD63 and a CD123 downward trend. Plasma TARC/CCL17 at baseline and post-stimulation cell supernatant histamine levels were similar in the three groups. CONCLUSIONS: In CF-ABPA, blood basophils are primed and hyperresponsive to Af allergen stimulation.Journal of cystic fibrosis: official journal of the European Cystic Fibrosis Society 05/2012; 11(6). DOI:10.1016/j.jcf.2012.04.008 · 3.48 Impact Factor