Comparison of basophil activation tests using CD63 or CD203c expression in patients with insect venom allergy.
ABSTRACT 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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ABSTRACT: Background: An aluminum hydroxide-adsorbed depot allergoid preparation of six-grass pollen allergens has been developed for short-term preseasonal immunotherapy in pollinosis. However, only limited knowledge exists about its immunological and clinical effects. The aim of this study was to evaluate the basophil response, which can explain early clinical findings of short-term preseasonal allergoid immunotherapy in allergic rhinitis. Methods: In a double-blind, placebo-controlled study, 31 patients allergic to grass pollens received one course of short-term preseasonal allergoid immunotherapy or placebo. Immunogenicity was assessed by the levels of specific IgG4, IgE antibodies and an allergen-induced CD203c basophil activation test. The primary clinical end point was the combined symptom and medication score/average combined score (ACS). Results: There was a 52.9% difference in ACS between the treatment and placebo groups in favor of immunotherapy (p = 0.01). Active treatment induced Phleum pratense-specific IgG4 and IgE antibodies (p < 0.05). A decrease in allergen-induced basophil activation at submaximal allergen concentrations was demonstrated at the end of immunotherapy and at the peak of the grass pollen season after immunotherapy. Conclusions: This study shows that grass pollen-allergic patients treated with one course of short-term preseasonal allergoid immunotherapy exhibit a decrease in allergen-induced basophil activation, an increase in allergen-specific IgG4 antibodies and early clinical improvement. © 2014 S. Karger AG, Basel.International Archives of Allergy and Immunology 08/2014; 164(3):237-245. DOI:10.1159/000365628 · 2.43 Impact Factor
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ABSTRACT: Background Approximately half of patients with chronic urticaria (CU) have functional autoantibodies against FceRIα or against IgE that induce histamine release from basophils and cutaneous mast cells. In addition to intracutaneous autologous serum skin test (ASST), more recently upregulation of CD63 and CD203c molecules on basophils has been proposed to detect the presence of autoantibodies in sera of CU patients. Objective Our aim was to assess the effect of serum from CU patients on basophil CD63 and CD203c expression and to correlate serum-induced basophil activation with ASST. Methods Sera were obtained from 128 patients with CU and 30 healthy controls. Patients or controls serum was incubated with atopic donor whole blood and activated basophils were identified by flow cytometry on basis of presence of CD63 or CD203c on high-expressing IgE positive cells. ASST was performed in all CU patients and 10 healthy controls. Results ASST was positive in 33.6% CU patients. Serum from 36.7% patients induced upregulation of CD63 while serum from 45.3% patients upregulated the CD203c molecule. There was a positive correlation between ASST and upregulation of CD63, but no correlation was observed between ASST and serum-induced CD203c. The sensitivity and specificity of the ASST in vitro assays were 55.8% and 72.9% for CD63; 55.8% and 60.0% for CD203 respectively. Conclusions Serum from CU patients upregulated both CD63 and CD203c molecules on blood basophils. A positive correlation between CD63 assay and ASST indicates the potential usefulness of this test for the diagnosis of the autoimmune form of CU.Central European Journal of Medicine 04/2014; 9(2). DOI:10.2478/s11536-013-0274-2 · 0.21 Impact Factor
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ABSTRACT: Challenges in in vitro allergy diagnostics lie in the development of accessible and reliable assays allowing identification of all offending allergens and cross-reactive structures. Flow-assisted analysis and quantification of in vitro activated basophils serves as a diagnostic instrument with increasing applications developed over the years. From the earliest days it was clear that the test could constitute a diagnostic asset in basophil-mediated hypersensitivity. However, utility of the basophil activation test should be reassessed regarding difficulties with preparation, characterization and validation of allergen extracts; availability and the potential of more accessible diagnostics. Today, the added value mainly lies in diagnosis of immediate drug hypersensitivity. Other potential indications are monitoring venom-immunotherapy and follow-up of natural history of food allergies. However, results in these nondiagnostic applications are preliminary. We review the most relevant clinical applications of the basophil activation test. Some personal comments and views about perspectives and challenges about flow-assisted allergy diagnosis are made.Expert Review of Clinical Immunology 10/2014; 10(10):1325-1335. DOI:10.1586/1744666X.2014.959498 · 3.34 Impact Factor