Correlation of allergen-specific T follicular helper cell counts with specific IgE levels and efficacy of allergen immunotherapy Yin Yao, MD, Cai-Ling.

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Correlation of allergen-specific T follicular helper cell counts with specific IgE levels and efficacy of allergen immunotherapy Yin Yao, MD, Cai-Ling Chen, MD, Nan Wang, MD, PhD, Zhi-Chao Wang, MD, Jin Ma, MD, PhD, Rong-Fei Zhu, MD, PhD, Xiao-Yan Xu, MD, Peng-Cheng Zhou, PhD, Di Yu, PhD, Zheng Liu, MD, PhD Journal of Allergy and Clinical Immunology Volume 142, Issue 1, Pages 321-324.e10 (July 2018) DOI: 10.1016/j.jaci.2018.03.008 Copyright © 2018 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 1 Increased numbers of circulating TFH2 cells in patients with AR. A, Percentages of circulating non-TFH and TFH cells among total CD4+ T cells in patients with AR (n = 27) and HC subjects (n = 28). B, Percentage of cell subsets among non-TFH and TFH cells in patients with AR (n = 27) and HC subjects (n = 28). C, TFH and non-TFH cells were cocultured with autologous naive B cells, and immunoglobulin levels were detected at day 8 (n = 8). D, Non-TFH and TFH cell subsets were cocultured with autologous naive B cells, and IgE levels were detected at day 8 (n = 6). E, TFH2 cells from patients with AR were cocultured with autologous naive B cells in the presence of blocking antibody or protein, and IgE levels were detected at day 8 (n = 6). In Fig 1, A and B, data are presented as means ± SDs. In Fig 1, C-E, data are presented as means ± SEMs. *P < .05 and **P < .01. Journal of Allergy and Clinical Immunology 2018 142, 321-324.e10DOI: (10.1016/j.jaci.2018.03.008) Copyright © 2018 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 2 Decrease in numbers of circulating allergen-specific IL-4+CXCR5+CD4+ TFH cells after AIT. A, Correlations between frequencies of Der p 1–specific IL-4+ non-TFH and TFH cells with D pteronyssinus–specific IgE levels in patients with AR. B, Changes in percentages of Der p 1–specific IL-4+ cells within non-TFH and TFH cells in patients with AR with (n = 22) and without (n = 20) AIT after treatment. Data are presented as means ± SDs. C, Correlations of reduction in numbers of Der p 1–specific IL-4+ non-TFH and TFH cells with CSMS improvement in patients with AR with and without AIT after treatment. *P < .05 and ***P < .001. Journal of Allergy and Clinical Immunology 2018 142, 321-324.e10DOI: (10.1016/j.jaci.2018.03.008) Copyright © 2018 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig E1 Peripheral TFH and non-TFH cells among total CD4+ T cells. A, Gating strategy for CXCR5− non-TFH cells and CXCR5+ TFH cells and their subsets in peripheral blood. FSC, Forward scatter; FV700, fixable viability stain 700; SSC, side scatter. B, Percentages of subsets of TFH and non-TFH cells in total CD4+ T cells (n = 28 for HC subjects and n = 27 for patients with AR). Data are presented as means ± SDs. *P < .05. Journal of Allergy and Clinical Immunology 2018 142, 321-324.e10DOI: (10.1016/j.jaci.2018.03.008) Copyright © 2018 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig E2 Immunoglobulin-inducing function of circulating TFH and non-TFH cells. A, Non-TFH and TFH cells from HC subjects or patients with AR were cocultured with autologous naive B cells, and immunoglobulin levels were detected at day 8 (n = 8). B, TFH and non-TFH cell subsets from HC subjects or patients with AR were cocultured with autologous naive B cells, and immunoglobulin levels were detected at day 8 (n = 6). C, TFH2 cells from patients with AR were cocultured with autologous naive B cells, and cytokine levels were detected at day 2 (n = 6). Data are presented as means ± SEMs. **P < .01. Journal of Allergy and Clinical Immunology 2018 142, 321-324.e10DOI: (10.1016/j.jaci.2018.03.008) Copyright © 2018 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig E3 Allergen-specific circulating TFH cells in patients with AR. A, Carboxyfluorescein succinimidyl ester (CFSE)–labeled circulating CXCR5− non-TFH and CXCR5+ TFH cells were cultured with autologous monocytes, which were pretreated with Der p 1 or staphylococcal enterotoxin B. Cell proliferation was analyzed by using flow cytometry at day 7 (n = 6). Representative histograms are shown. Data are expressed as means ± SEMs. B, Percentages of Der p 1–specific IL-4+ non-TFH and TFH cells within corresponding total non-TFH and TFH cells. Representative flow cytometric plots are shown. Data are presented as means ± SDs (n = 27 for patients with AR and n = 25 for HC subjects). *P < .05, **P < .01, and ***P < .001. Journal of Allergy and Clinical Immunology 2018 142, 321-324.e10DOI: (10.1016/j.jaci.2018.03.008) Copyright © 2018 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig E4 Changes in immunoglobulin levels and TH cell counts in patients with AR with or without subcutaneous AIT after treatment. A, Changes in CSMS. Data are expressed as means ± SDs. B, Changes in total IgE, D pteronyssinus–specific IgE, and D pteronyssinus–specific IgG4 levels. Data are expressed as means ± SEMs. C-E, Changes in TH cell (Fig E4, C), CXCR5− non-TFH cell subset (Fig E4, D), and CXCR5+ TFH cell subset counts (Fig E4, E; n = 22 for patients with AIT and n = 20 for patients without AIT). F, Correlations of reductions in D pteronyssinus–specific IgE levels with improvement in CSMS and decrease in Der p 1–specific IL-4+ TFH cell counts. Data are expressed as means ± SDs. *P < .05, **P < .01, and ***P < .001. Journal of Allergy and Clinical Immunology 2018 142, 321-324.e10DOI: (10.1016/j.jaci.2018.03.008) Copyright © 2018 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig E5 Patterns of chemokine receptor expression in Der p 1–specific IL-4+ TFH and Der p 1–specific IL-4+ non-TFH cells in patients with AR. Representative histograms are shown. Numbers in figures refer to percentages of positive cells (n = 2). Journal of Allergy and Clinical Immunology 2018 142, 321-324.e10DOI: (10.1016/j.jaci.2018.03.008) Copyright © 2018 American Academy of Allergy, Asthma & Immunology Terms and Conditions