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. 2019 Apr 5;10:431. doi: 10.3389/fimmu.2019.00431

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Copyright © 2019 Tebruegge, Ritz, Donath, Dutta, Forbes, Clifford, Zufferey, De Rose, Robins-Browne, Hanekom, Graham, Connell and Curtis.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Gating strategy. (A) Gating of CD3+ T cells, aggregate exclusion, followed by gating of single CD4+ and CD8+ T cells. (B) Threshold gates for CD4+ T cells producing IFN-γ, IL-2, TNF-α, or IL-17; a Boolean gating strategy was used to determine the proportion of polyfunctional T-cells. Gates for cytokine-producing CD4+ T cells were set on the unstimulated sample (upper panel) and transposed onto the antigen-stimulated samples (in this example PPD; lower panel) in each individual participant.