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Flow-assisted quantification of in vitro activated basophils in the diagnosis of wasp venom allergy and follow-up of wasp venom immunotherapy

  1. D. G. Ebo,
  2. M. M. Hagendorens,
  3. A. J. Schuerwegh,
  4. L. M.-N. Beirens,
  5. C. H. Bridts,
  6. L. S. De Clerck,
  7. W. J. Stevens

Article first published online: 16 NOV 2006

DOI: 10.1002/cyto.b.20142

Issue

Cytometry Part B: Clinical Cytometry

Cytometry Part B: Clinical Cytometry

Volume 72B, Issue 3, pages 196–203, May 2007

Additional Information

How to Cite

Ebo, D. G., Hagendorens, M. M., Schuerwegh, A. J., Beirens, L. M.-N., Bridts, C. H., De Clerck, L. S. and Stevens, W. J. (2007), Flow-assisted quantification of in vitro activated basophils in the diagnosis of wasp venom allergy and follow-up of wasp venom immunotherapy. Cytometry Part B: Clinical Cytometry, 72B: 196–203. doi: 10.1002/cyto.b.20142

Author Information

  1. Department of Immunology, Allergology and Rheumatology, University of Antwerp, Belgium

*Correspondence: W. J. Stevens, Immunologie–Allergologie–Reumatologie, Universiteit Antwerpen, Campus Drie Eiken, Universiteitsplein 1, 2610 Antwerpen (Wilrijk), Belgium

Publication History

  1. Issue published online: 9 APR 2007
  2. Article first published online: 16 NOV 2006
  3. Manuscript Accepted: 7 JUL 2006
  4. Manuscript Revised: 21 JUN 2006
  5. Manuscript Received: 19 DEC 2005

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Keywords:

  • flow cytometry;
  • CD63;
  • immunotherapy;
  • hymenoptera venom

Abstract

Background:

Correct identification of the culprit venom is a prerequisite for specific venom immunotherapy (VIT). Despite the efficacy of VIT, issues as how to monitor treatment and when to discontinue maintenance therapy remain to be established.

Methods:

To evaluate diagnostic performances of the basophil activation test (BAT) in wasp venom allergy, 80 patients with a definite history of wasp venom anaphylaxis (systemic reactors) and 14 wasp-stung asymptomatic controls (stung controls) were enrolled. Venom-induced basophil activation was analyzed flow cytometrically by double-labeling with anti-IgE and anti-CD63. Results were compared to wasp IgE levels and results of a venom skin test (VST). To establish whether the BAT constitutes a candidate marker to monitor VIT, the BAT was repeated in 22 patients on the 5th day of a build-up course and after 6 months of maintenance VIT. Whether the BAT could contribute in the decision of discontinuing VIT was assessed in a cross-sectional analysis in 30 patients receiving treatment for 3 years.

Results:

Comparison between systemic reactors and stung controls revealed a sensitivity of 86.4% and specificity of 100% for venom IgE, and sensitivity of 81.8% for VST, respectively. In contrast to stung controls, patients demonstrated dose-dependent venom-induced basophil activation. The BAT attained a sensitivity of 83.8% and specificity of 100%. At the end of the build-up course, no effect of VIT on the BAT was demonstrable. When the BAT was repeated after 6 months of treatment, submaximal stimulation of the cells demonstrated a significant decreased CD63 expression (P < 0.04). Patients having VIT for 3 years also demonstrated significantly lower venom-induced CD63 expression (P < 0.001). After 3 years, 60% of the patients had a negative BAT for submaximal stimulation of the cells whereas only 17.9% of the patients had negativation of wasp IgE.

Conclusions:

The BAT is a reliable instrument for the diagnosis of wasp venom anaphylaxis and might constitute an instrument to monitor wasp VIT. © 2006 International Society for Analytical Cytology

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