Abstract
BackgroundFood allergy is an increasing public health issue and the most common cause of life-threatening anaphylactic reactions. Conventional allergy tests assess for the presence of allergen-specific IgE, significantly overestimating the rate of true clinical allergy and resulting in overdiagnosis and adverse effect on health-related quality of life.ObjectiveTo undertake initial validation and assessment of a novel diagnostic tool, we used the mast cell activation test (MAT).MethodsPrimary human blood-derived mast cells (MCs) were generated from peripheral blood precursors, sensitized with patients' sera, and then incubated with allergen. MC degranulation was assessed by means of flow cytometry and mediator release. We compared the diagnostic performance of MATs with that of existing diagnostic tools to assess in a cohort of peanut-sensitized subjects undergoing double-blind, placebo-controlled challenge.ResultsHuman blood-derived MCs sensitized with sera from patients with peanut, grass pollen, and Hymenoptera (wasp venom) allergy demonstrated allergen-specific and dose-dependent degranulation, as determined based on both expression of surface activation markers (CD63 and CD107a) and functional assays (prostaglandin D2 and β-hexosaminidase release). In this cohort of peanut-sensitized subjects, the MAT was found to have superior discrimination performance compared with other testing modalities, including component-resolved diagnostics and basophil activation tests. Using functional principle component analysis, we identified 5 clusters or patterns of reactivity in the resulting dose-response curves, which at preliminary analysis corresponded to the reaction phenotypes seen at challenge.ConclusionThe MAT is a robust tool that can confer superior diagnostic performance compared with existing allergy diagnostics and might be useful to explore differences in effector cell function between basophils and MCs during allergic reactions.
Highlights
Food allergy is an increasing public health issue and the most common cause of life-threatening anaphylactic reactions
We developed a robust and reproducible mast cells (MCs)-based assay to improve the diagnosis of IgE-mediated allergy using human MCs derived from human progenitor cells. human blood-derived mast cells (hMCs) sensitized with sera from patients with peanut, grass pollen, and Hymenoptera allergy demonstrated allergen-specific and dosedependent degranulation by using both expression of surface activation markers (CD63 and CD107a) and functional assays (PGD2 and b-hexosaminidase release)
The mast cell activation test (MAT) is a very sensitive assay, with significant levels of surface expression of CD63 activation markers after stimulation with peanut at concentrations up to 2-log lower than that required for the basophil activation test (BAT).[15]
Summary
Food allergy is an increasing public health issue and the most common cause of life-threatening anaphylactic reactions. We compared the diagnostic performance of MATs with that of existing diagnostic tools to assess in a cohort of peanut-sensitized subjects undergoing double-blind, placebocontrolled challenge. Results: Human blood-derived MCs sensitized with sera from patients with peanut, grass pollen, and Hymenoptera (wasp venom) allergy demonstrated allergen-specific and dosedependent degranulation, as determined based on both expression of surface activation markers (CD63 and CD107a) and functional assays (prostaglandin D2 and b-hexosaminidase release). In this cohort of peanut-sensitized subjects, the MAT was found to have superior discrimination performance compared with other testing modalities, including componentresolved diagnostics and basophil activation tests. Conclusion: The MAT is a robust tool that can confer superior diagnostic performance compared with existing allergy diagnostics and might be useful to explore differences in effector cell function between basophils and MCs during allergic reactions. (J Allergy Clin Immunol 2018;142:485-96.)
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