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An integrated framework using high‐dimensional mass cytometry and fluorescent flow cytometry identifies discrete B cell subsets in patients with red meat allergy
Author(s) -
Cox Kelly M.,
Commins Scott P.,
Capaldo Brian J.,
Workman Lisa J.,
PlattsMills Thomas A. E.,
Amir Elad D.,
Lannigan Josephine A.,
Schuyler Alexander J.,
Erickson Loren D.
Publication year - 2019
Publication title -
clinical and experimental allergy
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.462
H-Index - 154
eISSN - 1365-2222
pISSN - 0954-7894
DOI - 10.1111/cea.13322
Subject(s) - flow cytometry , elispot , immunology , immunoglobulin d , immunoglobulin e , allergy , peripheral blood mononuclear cell , cytometry , b cell , food allergy , mass cytometry , biology , isotype , antibody , t cell , phenotype , in vitro , immune system , genetics , gene , monoclonal antibody
Summary 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 CD 27. These B cell subsets shared higher IgD and lower IgM expression levels coupled with CXCR 4, CCR 6 and CD 25 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.