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http://dx.doi.org/10.5045/br.2018.53.4.299

Neutrophil oxidative burst as a diagnostic indicator of IgG-mediated anaphylaxis  

Won, Dong Il (Department of Clinical Pathology, School of Medicine, Kyungpook National University)
Kim, Sujeong (Division of Allergy and Infectious Diseases, Department of Internal Medicine, School of Medicine, Kyungpook National University)
Lee, Eun Hee (Green Cross Reference Laboratory)
Publication Information
BLOOD RESEARCH / v.53, no.4, 2018 , pp. 299-306 More about this Journal
Abstract
Background IgG-mediated anaphylaxis occurs after infusion of certain monoclonal antibody-based therapeutics. New in vitro tests are urgently needed to diagnose such reactions. We investigated whether allergens trigger neutrophil oxidative burst (OB) and if neutrophil OB occurs due to allergen-specific IgG (sIgG). Methods Neutrophil OB was measured by dihydrorhodamine 123 flow cytometry using a leukocyte suspension spiked with a very small patch of the allergen crude extract, Dermatophagoides farinae (Der f). The mean fluorescence intensity ratio of stimulated to unstimulated samples was calculated as the neutrophil oxidative index (NOI). Results The Der f-specific NOI (Der f-sNOI) showed a time-dependent increase after Der f extract addition. At 15 min activation, higher Der f-sIgG levels were associated with lower Der f-sNOI values in 31 subjects (P<0.05). This inverse relationship occurs due to the initial blocking effect of free Der f-sIgG. Additionally, neutrophil OB was nearly absent (Der f-sNOI of -1) in two cases: a subject with undetectable Der f-sIgG levels and washed leukocyte suspensions deprived of Der f-sIgG. Conclusion Allergens can trigger neutrophil OB via preexisting allergen-sIgG. Neutrophil OB can be easily measured in a leukocyte suspension spiked with the allergen. This assay can be used to diagnose IgG-mediated anaphylaxis.
Keywords
Neutrophils; Oxidative burst; Anaphylaxis; Immunoglobulin G; Dihydrorhodamine 123;
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