Basophil activation in insect venom allergy: comparison of an established test using liquid reagents with a test using 5-color tubes with dried antibody reagents

BMC Immunol. 2024 Apr 27;25(1):23. doi: 10.1186/s12865-024-00616-0.

Abstract

Background: Flow cytometry-based basophil activation tests (BAT) have been performed with various modifications, differing in the use of distinct identification and activation markers. Established tests use liquid reagents while a new development involves the use of tubes with dried antibody reagents. The aim of this pilot study was to compare these two techniques in patients with insect venom allergy.

Methods: Seventeen patients with an insect venom allergy were included in the study. The established "BAT 1" utilizes conventional antibody solutions of anti-CCR3 for basophil identification and anti-CD63 to assess basophil activation, whereas "BAT 2" uses dried anti-CD45, anti-CD3, anti-CRTH2, anti-203c and anti-CD63 for identification and activation measurement of basophils. Negative and positive controls as well as incubations with honey bee venom and yellow jacket venom at three concentrations were performed.

Results: Seven patients had to be excluded due to low basophil counts, high values in negative controls or negative positive controls. For the remaining 10 patients the overall mean (± SD) difference in activated basophils between the two tests was 0.2 (± 12.2) %P. In a Bland-Altman plot, the limit of agreement (LoA) ranged from 24.0 to -23.7. In the qualitative evaluation (value below/above cut-off) Cohen's kappa was 0.77 indicating substantial agreement. BAT 2 took longer to perform than BAT 1 and was more expensive.

Conclusion: The BAT 2 technique represents an interesting innovation, however, it was found to be less suitable compared to an established BAT for the routine diagnosis of insect venom allergies.

Keywords: Basophil activation test; Dried antibody reagents; Insect venom allergy; Liquid reagents.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adolescent
  • Adult
  • Aged
  • Animals
  • Antibodies / immunology
  • Arthropod Venoms / immunology
  • Basophil Degranulation Test / methods
  • Basophils* / immunology
  • Bee Venoms / immunology
  • Female
  • Flow Cytometry* / methods
  • Humans
  • Hypersensitivity / diagnosis
  • Hypersensitivity / immunology
  • Insect Bites and Stings / diagnosis
  • Insect Bites and Stings / immunology
  • Male
  • Middle Aged
  • Pilot Projects
  • Venom Hypersensitivity
  • Young Adult

Substances

  • Arthropod Venoms
  • Bee Venoms
  • Antibodies