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. 2021 Nov 23;12:727300. doi: 10.3389/fimmu.2021.727300

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Copyright © 2021 Silva, Sundling, Folkesson, Fröberg, Nobrega, Canto-Gomes, Chambers, Lakshmikanth, Brodin, Bruchfeld, Nigou, Correia-Neves and Källenius

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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Cytokine production by stimulated NK cells. (A) The number of cytokine-producing NK cells per 1000 total CD45⁺ cells for each stimulation at 24 h with the background (unstimulated) cytokine-production removed. (B) Cluster analysis of IFN-γ secreting cells with clusters 1, 6, 7, 13, 19, 21, 22, 23, and 25 corresponding to NK cells. (C) Clusters significantly affected by stimulation with (D) cluster histograms indicating CD56 expression and GrzB and IL-17A secretion. (E) Cluster analysis of IL-2 secreting cells with cluster 1 and 9 corresponding to NK cells. (F) Clusters significantly affected by stimulation with (G) cluster dot plots showing CD56, CD57, and CD27 expression and histograms indicating IL-6 production. (H) Cluster analysis of GM-CSF secreting cells with cluster 4 and 16 corresponding to NK cells. (I) Cluster 16 significantly affected by stimulation. (J) Cluster’s dot plots showing CD56, CD57, and CD27 expression and histogram indicating IFN-γ secretion. (K) Cluster analysis of IL-6 secreting cells with cluster 1 and 9 corresponding to NK cells. (L) Clusters significantly affected by stimulation (left) and comparison of PIM stimulation on donors with active TB (ATB), latent TB (LTB) and healthy controls (HC) in cluster 12 (right). (M) Cluster’s dot plots showing CD56, CD57, and CD27 expression. Numbers in dot plots indicate the percentage within the cluster. Statistical differences between stimulations in (A, F, I) were evaluated by Friedman’s test with Dunnet’s posttest, comparisons within multiple clusters (C, L left) were evaluated by a matched-pair two-way ANOVA with Geissner-Greenhouse correction followed by Tukey’s posttest (n=15/stimulation). Groups (ATB/LTB/HC) (L, right) were compared using Kruskal-Wallis with Dunn’s posttest (n=5/group) *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001.