Detection of granulocyte-reactive antibodies: a comparison of different methods

Vox Sang. 2015 Apr;108(3):287-93. doi: 10.1111/vox.12227. Epub 2014 Dec 30.

Abstract

Background and objectives: Granulocyte-reactive antibodies can cause autoimmune and neonatal immune neutropenias as well as transfusion-related acute lung injury. The classical antibody-detection methods granulocyte aggregation test (GAT), granulocyte immunofluorescence test (GIFT) and monoclonal antibody-specific immobilization of granulocyte antigens (MAIGA) are time-consuming and technically challenging. In recent years, flow cytometric white blood cell immunofluorescence test (Flow-WIFT) and the microbeads assay LabScreen® Multi have emerged and are still subject of evaluation. These serological tests were compared on a screening and specification level.

Materials and methods: For screening, the combination of GAT/GIFT was compared to Flow-WIFT testing 333 samples. Positive samples were further analysed with MAIGA and LabScreen® Multi.

Results: Granulocyte aggregation test/GIFT detected 77 positive samples, Flow-WIFT found 108 granulocyte-reactive samples. Six Samples were only positive in GAT/GIFT, and 37 samples were only positive in Flow-WIFT (κ = 0.682). Antibody specification with MAIGA and the microbeads assay confirmed granulocyte-reactivity in 83 cases with 70 matching results (κ = 0.742). However, out of six detected human neutrophil antigen (HNA) reactivities only two specificities matched in both assays.

Conclusion: Flow-WIFT may be a valuable addition to GIFT for granulocyte-reactive antibody screening. MAIGA remains the most reliable laboratory method for antibody specification.

Keywords: flow cytometry; granulocyte immunology; granulocyte-reactive antibodies; human neutrophil antigens; microbeads assay.

Publication types

  • Comparative Study

MeSH terms

  • Agglutination Tests / methods*
  • Antibodies, Monoclonal / immunology*
  • Antibody Specificity
  • Granulocytes / immunology*
  • Humans

Substances

  • Antibodies, Monoclonal