An integrated framework using high‐dimensional mass cytometry and fluorescent flow cytometry identifies discrete B cell subsets in patients with red meat allergy

Abstract

Background

B cells play a critical role in the development and maintenance of food allergy by producing allergen‐specific IgE. Despite the importance of B cells in IgE‐mediated food allergy, the identity of sIgE‐producing human B cells and how IgE is regulated are poorly understood.

Objective

To identify the immunophenotypes of circulating B cells associated with the production of galactose‐alpha‐1,3‐galactose specific IgE production in patients with red meat allergy.

Methods

B cells in PBMC samples obtained from 19 adults with physician‐diagnosed red meat allergy and 20 non‐meat allergic healthy controls were assessed by mass cytometry along with a bioinformatics analysis pipeline to identify discrete B cell phenotypes that associated with serum sIgE. Fluorescent flow cytometry was then applied to sort purify discrete B cell subsets, and B cells were functionally evaluated on an individual cell level for the production of sIgE by ELISPOT.

Results

Discrete B cell phenotypes abundant in meat allergic subjects compared to non‐meat allergic controls were found in peripheral blood that do not share typical characteristics of classical isotype‐switched memory B cells that express high levels of CD27. These B cell subsets shared higher IgD and lower IgM expression levels coupled with CXCR4, CCR6 and CD25 expression. In vitro polyclonal stimulation of purified B cell subsets from meat allergic subjects demonstrated that these subsets were enriched for cells induced to secrete sIgE.

Conclusions and Clinical Relevance

Circulating B cells display increased abundance of discrete B cell subsets in meat allergic subjects. This observation, coupled with the capacity of individual B cell subsets to produce sIgE following activation, implicates these novel B cell phenotypes in promoting IgE in meat allergy.

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