Cytometric analysis reveals an association between allergen-responsive natural killer cells and human peanut allergy
Xiaoying Zhou, … , Rosemarie H. DeKruyff, Kari C. Nadeau
Xiaoying Zhou, … , Rosemarie H. DeKruyff, Kari C. Nadeau
Published October 17, 2022
Citation Information: J Clin Invest. 2022;132(20):e157962. https://doi.org/10.1172/JCI157962.
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Research Article Immunology

Cytometric analysis reveals an association between allergen-responsive natural killer cells and human peanut allergy

Abstract

Food allergies are a leading cause of anaphylaxis, and allergen-specific immune responses in both the innate and the adaptive immune system play key roles in its pathogenesis. We conducted a comprehensive phenotypic and functional investigation of immune cell responses from nonallergic (NA) and peanut allergic (PA) participants cultured with media alone or peanut protein and found, surprisingly, that NK cell activation was strongly associated with the immune response to allergen in PA participants. Peanut-responsive NK cells manifested a distinct expression pattern in PA participants compared with NA participants. Allergen-activated NK cells expressed both Th2 and immune regulatory cytokines, hinting at a potential functional role in mediating and regulating the Th2 allergic response. Depletion of CD3+ T cells attenuated the response of NK cells to peanut-allergen stimulation, suggesting that peanut-responsive NK cells are T cell dependent. We also showed that oral immune therapy was associated with decreased NK responses to peanut allergen stimulation in vitro. These results demonstrate that NK cells are associated with the food-allergic immune response, and the magnitude of this mobilized cell population suggests that they play a functional role in allergic immunity.

Authors

Xiaoying Zhou, Wong Yu, Diane M. Dunham, Jackson P. Schuetz, Catherine A. Blish, Rosemarie H. DeKruyff, Kari C. Nadeau

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Figure 2

The magnitude of peanut-dependent expansion is greater in CD69+ NK cells than in effector memory Th2 cells.

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The magnitude of peanut-dependent expansion is greater in CD69+ NK cells...
(A) Representative mass cytometry plots show the gating strategy for memory Th2 cells (CD14–CD19–CD3+CD4+CD45RA–CXCR3–CCR4+CCR6–). (B) FlowSOM-based clustering analysis was performed on gated memory Th2 cells from PBMCs from NA and PA participants cultured with media alone or peanut protein (n = 26 for NA PBMCs cultured with or without peanut protein, n = 22 for PA PBMCs cultured with or without peanut protein). Heatmap displays the median expression levels of cell-surface markers in each cluster. The marker expression values were arcsinh transformed with a cofactor of 5. The bar graph shows each cluster as percentage of memory Th2 cells. (C) UMAP representation of 96,000 randomly selected cells (1,000 per sample; total 96 samples, n = 26 for NA PBMCs cultured with or without peanut protein, n = 22 for PA PBMCs cultured with or without peanut protein); clusters from the FlowSOM analysis are indicated by color. (D) The frequencies of 5 memory Th2 cell subsets as a percentage of total PBMCs from NA (n = 26) and PA (n = 22) participants cultured with or without peanut stimulation. Each pair of points connected by a line represents 1 participant. Box plots indicate the IQR and median; whiskers extend to the farthest data point within a maximum of 1.5 × IQR. Paired sample sets were analyzed using a 2-sided Wilcoxon signed rank test. Unpaired sample sets were analyzed using a 2-sided Wilcoxon rank sum test. P values were adjusted for multiple comparisons using the Bonferroni approach to control the FDR. FDR-adjusted P < 0.05 were considered significant. The stars indicate the FDR-adjusted P values, ***P < 0.001 and **P < 0.01. (E) Average frequencies of activated NK cell subsets and activated EM CD4+ cell subsets as a percentage of total PBMCs from PA participants cultured with peanut protein.