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. 2018 Nov 15;8:16904. doi: 10.1038/s41598-018-35161-5

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© The Author(s) 2018

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fungi specific T cells produce IL-17 in an age dependent manner. CD4⁺CD45RA⁺CD31⁺ T cells from neonates, infants, children, and adults were co-cultured with monocytes pulsed with C. albicans- or A. fumigatus-lysates. (A,B) The frequency of T cells expressing signature Th17 molecules IL-17 (A) and RORγt (B) were analysed by flow cytometry at day 3 (upper Panel) and day 6 after stimulation (lower panel) (C) Bar graph representing the ELISPOT analysis of the quantitative IL-17, produced by the T cells from neonates and adults which were either stimulated or not for 3 days as described in (A). (D) Determination of IL-17A cytokine release of CD4⁺CD45RA⁺ T cells of neonates, infants and children or adults by LegendPlex which were either stimulated or not for 3 days as described in (A). (E) Frequency of T cells expressing intracellular IL-17 (white), IL-4 (black) and both IL-17/IL-4 (grey) were measured by flow cytometry after 6 days of stimulation, analysed by boolean gating and shown as different fractions of cytokine expressing cells in a stacked bar chart. (F) Bar graph showing IL-17 expression by CD4⁺CD45RA⁺CD31⁺ T cells of neonates, adults, and infants of 0.5-2 years old, stimulated for 6 days as described in (A) in the presence or absence of neutralizing antibodies for IL-1ß, IL-6 or both. Cumulative results are shown and each dot in (A–D) and (F) represent a different donor. The error bars in figures denote ± SD. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001, as determined by one-way Anova with Tukey post hoc test (A-C, F) or Kruskal Wallis with Dunn’s post hoc test (D).