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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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Identification of age-dependent fungi specific multifunctional Th1/Th17 cells. (A) CD4⁺CD45RA⁺CD31⁺ T cells from neonates, infants and children of different age groups, and adults were co-cultured with monocytes pulsed with C. albicans (left panel) or A. fumigatus (right panel). The frequency of T cells expressing intracellular IL-17 (white), IFNγ (black) and both IL-17/IFNγ (grey) were determined by flow cytometry at the indicated time points, analysed by boolean gating and shown as different fractions of cytokine expressing cells in a stacked bar chart. (B) Frequency of naïve (CD4⁺CD45RA⁺CD31⁺) or memory (CD4⁺CD45RO⁺) T cells of adults expressing intracellular IL-17 (white), IFNγ (black) and both IL-17/IFNγ (grey) were determined by flow cytometry at the indicated time points and analysed as described in (A). (C) Frequency of CD4⁺CD45RA⁺CD31⁺ T cells from neonates and adults expressing Th1 and Th17 transcription factors T-bet, RORγt and both T-bet as well as RORγt following stimulation with C. albicans (orange bars) or A. fumigatus (blue bars) for 3 and 6 days as described in (A). Cumulative results are shown and each dot represents 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 Kruskal Wallis test with Dunn’s post hoc test.