Der p 1–induced CD4+FOXP3+GATA3+ T cells have suppressive properties and contribute to the polarization of the TH2-associated response Lieke Reubsaet,

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Der p 1–induced CD4+FOXP3+GATA3+ T cells have suppressive properties and contribute to the polarization of the TH2-associated response Lieke Reubsaet, MSc, MD, Jenny Meerding, BSc, Rianne Giezeman, MSc, MD, Ismé de Kleer, MD, PhD, Bert Arets, MD, PhD, Berent Prakken, MD, PhD, Jeffrey Beekman, PhD, Femke van Wijk, PhD Journal of Allergy and Clinical Immunology Volume 132, Issue 6, Pages 1440-1444.e2 (December 2013) DOI: 10.1016/j.jaci.2013.06.028 Copyright © 2013 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 1 Der p 1 and Bet v 1 induce CD4+FOXP3+GATA3+ cells in pediatric HDM-sensitized subjects. A, CD4+ T-cell proliferation after culture with Der p 1 (n = 4) or Bet v 1 (n = 4). B, FOXP3 and GATA3 protein expression in CD4+ T cells of HDM- or birch-sensitized children (n = 8). C, Representative FACS plot and FOXP3 and GATA3 protein expression in proliferating and nonproliferating CD4+ T cells of HDM-sensitized children. D, Time course of FOXP3 and GATA3 protein expression in CD4+ T cells of HDM-sensitized children (n = 4). Data shown are after 7 days of PBMC culture, unless otherwise indicated (mean ± SEM). DN, Double negative; DP, double positive; SP, single positive. *P < .05 compared with day 0 or between groups, as indicated. Journal of Allergy and Clinical Immunology 2013 132, 1440-1444.e2DOI: (10.1016/j.jaci.2013.06.028) Copyright © 2013 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 2 Phenotypic and functional characterization of different cell populations after culture of PBMCs with Der p 1. A and B, Cell-surface marker expression (Fig 2, A) and cytokine production (Fig 2, B) of different populations after 7 days of culture of PBMCs from HDM-sensitized children (n = 4-8). C, Representative dot plot showing enrichment of the FOXP3/GATA3 double-positive cell population used in suppression assays. D and E, Level of suppression of CD4+ and CD8+ T-cell proliferation (Fig 2, D) and cytokines (Fig 2, E) by FOXP3/GATA3 double-positive enriched CD4+ T cells and ex vivo sorted natural Treg cells (nTreg) cultured in a 1:8 ratio with CFSE-labeled PBMCs. The level of suppression of PBMCs is set to 0 (mean ± SEM). DN, Double negative; DP, double positive; SP, single positive. *P < .05 compared with double-negative cells or between groups, as indicated. Journal of Allergy and Clinical Immunology 2013 132, 1440-1444.e2DOI: (10.1016/j.jaci.2013.06.028) Copyright © 2013 American Academy of Allergy, Asthma & Immunology Terms and Conditions