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. 2015 May 14;46(1):52. doi: 10.1186/s13567-015-0182-3

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© Talker et al.; licensee BioMed Central. 2015

This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly credited. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.

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Kinetics of FLUAVsw-specific IFN-γ-/TNF-α-/IL-2-producing CD4 ⁺ T cells. Intracellular cytokine staining of defrosted PBMCs was performed following overnight in vitro restimulation with FLUAVsw (infection strain, MOI = 0.1; 18 h). Mock-incubated cultures served as negative controls. (A) CD4⁺ T cells were gated and analyzed for production of IFN-γ, TNF-α and IL-2. Contour plots show combinations of cytokines for selected time points following FLUAVsw infection. Exemplary data of animal #9 is shown. (B) Boolean gating was applied in order to identify single-, double-, and triple-cytokine-producing CD4⁺ T cells. Single- (dark grey), double- (orange) and triple- (red) cytokine-producing cells are shown as percent of total CD4⁺ T cells for all six infected animals in the time course following FLUAVsw infection.