Allergen-specific immunotherapy modulates the balance of circulating Tfh and Tfr cells Véronique Schulten, PhD, Victoria Tripple, BSc, Grégory Seumois,

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Presentation transcript:

Allergen-specific immunotherapy modulates the balance of circulating Tfh and Tfr cells Véronique Schulten, PhD, Victoria Tripple, BSc, Grégory Seumois, PhD, Yu Qian, PhD, Richard H. Scheuermann, PhD, Zheng Fu, PhD, Michela Locci, PhD, Sandy Rosales, BSc, Pandurangan Vijayanand, MD, Alessandro Sette, PhD, Rafeul Alam, MD, Shane Crotty, PhD, Bjoern Peters, PhD Journal of Allergy and Clinical Immunology Volume 141, Issue 2, Pages 775-777.e6 (February 2018) DOI: 10.1016/j.jaci.2017.04.032 Copyright © 2017 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 1 AIT-treated patients exhibit reduced Tfh frequencies compared with untreated allergic and nonallergic controls. A, Representative Tfh cell staining in an allergic, AIT-treated, and nonallergic patient. Dashed boxes indicate the total Tfh (black), CXCR5lo (blue), and CXCR5hi (red) populations. B, Tfh cell percentage in allergic (n = 25), AIT-treated (n = 32) and nonallergic (n = 13) patients. C, Quantification of CXCR5 gene expression in CXCR5hi and CXCR5lo sorted cells. D, MA plots comparing fold changes of gene expression between CXCR5hi and CXCR5lo cells versus average gene expression. Genes with significant adjusted P values are colored in red. E, Heatmap of differentially expressed genes in CXCR5hi versus CXCR5lo samples from allergic and AIT-treated individuals. Journal of Allergy and Clinical Immunology 2018 141, 775-777.e6DOI: (10.1016/j.jaci.2017.04.032) Copyright © 2017 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 2 Protein expression profiles of selected markers in CXCR5hi versus CXCR5lo cells and the effect of TCR stimulation on CXCR5 expression. A, Quantification of protein expression of markers shown to be differentially expressed in CXCR5hi versus CXCR5lo cells as determined by RNAseq. B, Fold change of Helios and Granzyme A expression in CXCR5loFOXP3+ versus CXCR5loFOXP3− cells. C, Median fluorescent intensity (MFI) for CXCR5 expression in Tfr and Tfh cells. D, CXCR5 expression in sorted Tfh cells after 5 days of culture in medium alone or in the presence of anti-CD3/28 beads with or without anti–IL-2. E, Percentage of Tfr cells (Foxp3+Helios+) in total CXCR5+ cells after culture in medium or with anti-CD3/28 beads with or without anti–IL-2. Statistical analysis was done by Wilcoxon matched-pairs signed rank test (1-sided) (n = 46). ∗P < .05, ∗∗P < .01, and ∗∗∗∗P < .0001. Journal of Allergy and Clinical Immunology 2018 141, 775-777.e6DOI: (10.1016/j.jaci.2017.04.032) Copyright © 2017 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig E1 Automated gating analysis reveals reduced CXCR5 expression in AIT-treated patients compared with untreated allergic and nonallergic controls. A, Representative dotplots showing Tfh cells in an allergic, AIT-treated, and nonallergic patient. B, Percentage Tfh in allergic (n = 24), AIT-treated (n = 32), and nonallergic (n = 13) patients. Statistical analysis was performed using the Mann-Whitney test, 1-tailed. **P < .01. Journal of Allergy and Clinical Immunology 2018 141, 775-777.e6DOI: (10.1016/j.jaci.2017.04.032) Copyright © 2017 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig E2 Cytokine production after PMA/ionomycin stimulation in Tfr cells ex vivo. Intracellular cytokine staining for IL-10, IFN-γ, IL-4, and IL-13 after 6 hours of stimulation with PMA/ionomycin was performed in Tfr cells from 4 healthy donors. Percentage of cytokine-producing cells out of the total CD4 subset is shown. Journal of Allergy and Clinical Immunology 2018 141, 775-777.e6DOI: (10.1016/j.jaci.2017.04.032) Copyright © 2017 American Academy of Allergy, Asthma & Immunology Terms and Conditions