Hidradenitis Suppurativa Is Characterized by Dysregulation of the Th17:Treg Cell Axis, Which Is Corrected by Anti-TNF Therapy Barry Moran, Cheryl M.

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Hidradenitis Suppurativa Is Characterized by Dysregulation of the Th17:Treg Cell Axis, Which Is Corrected by Anti-TNF Therapy Barry Moran, Cheryl M. Sweeney, Rosalind Hughes, Anna Malara, Shivashini Kirthi, Anne-Marie Tobin, Brian Kirby, Jean M. Fletcher Journal of Investigative Dermatology Volume 137, Issue 11, Pages 2389-2395 (November 2017) DOI: 10.1016/j.jid.2017.05.033 Copyright © 2017 The Authors Terms and Conditions

Figure 1 CD4 T cells infiltrating HS skin secrete multiple proinflammatory cytokines, with a Th17 skewed profile, and exhibit a Treg:Th17 cell imbalance. Cells isolated from the skin and blood from healthy control subjects or HS patients were stimulated with phorbol 12-myristate 13-acetate and ionomycin, stained with surface and intracellular markers, and analyzed by flow cytometry. CD4 T cells were gated according to the strategy shown in Supplementary Figure S1a. Plots show the frequency of (a) cytokine-expressing cells, (b) Th subsets, (c) Treg cells, and Th17:Treg cell ratio in skin and blood of healthy control subjects and HS patients. Statistical significance is only between skin samples or PBMC samples, using one-way ANOVA with Newman-Keuls multiple comparison test. ∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.001. C, healthy control subjects; HS, hidradenitis suppurativa; L, lesional skin; P, perilesional skin; PBMC, peripheral blood mononuclear cell; Th, T helper cell; TNF, tumor necrosis factor; Treg, regulatory T cell; U, uninvolved skin. Journal of Investigative Dermatology 2017 137, 2389-2395DOI: (10.1016/j.jid.2017.05.033) Copyright © 2017 The Authors Terms and Conditions

Figure 2 A significant reduction in Th17 cells and correction of the Th17:Treg ratio are seen in HS patients treated with anti-TNF. Cells isolated from the skin from healthy control subjects or HS patients who were treatment naïve or treated with anti-TNF were processed and analyzed as per Figure 1. Plots show the frequency of (a) cytokine-expressing cells, (b) Th subsets, (c) Treg cells, and Th17:Treg cell ratio in skin and blood of healthy control subjects and HS patients who were untreated or treated with anti-TNF. Statistical significance is only between skin samples or PBMC samples and was determined by one-way analysis of variance with Newman-Keuls multiple comparison test. ∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.001. HS, hidradenitis suppurativa; L, lesional skin; P, perilesional skin; PBMC, peripheral blood mononuclear cell; Th, T helper cell; TNF, tumor necrosis factor; Treg, regulatory T cell; U, uninvolved skin. Journal of Investigative Dermatology 2017 137, 2389-2395DOI: (10.1016/j.jid.2017.05.033) Copyright © 2017 The Authors Terms and Conditions

Figure 3 Reduction in cytokine polyfunctionality of CD4 T cells in anti-TNF–treated HS patient skin. Cells were processed and analyzed as in Figure 1. The cytokine expression of CD4 T cells from lesional, perilesional, and uninvolved skin from HS patients who were anti-TNF–treated or –naïve, as well as healthy control skin, was analyzed using SPICE software. The pie charts represent the average frequencies of CD4 T cells producing every combination of cytokines for multiple donors. The segments within the pie chart denote populations producing different combinations of cytokines and are heatmap coded to indicate increasing cytokine production. The size of the pie segment correlates to the frequency of the population. The arcs indicate the cytokine produced by that proportion of cells. HS, hidradenitis suppurativa; TNF, tumor necrosis factor. Journal of Investigative Dermatology 2017 137, 2389-2395DOI: (10.1016/j.jid.2017.05.033) Copyright © 2017 The Authors Terms and Conditions