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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 CD33⁺ myeloid cells. (A) The number of cytokine positive CD33⁺ myeloid cells per 1000 total CD45⁺ cells for each stimulation at 24 h with the background (unstimulated) cytokine-production removed. (B) Cluster analysis of IL-2 secreting cells with cluster 3, 4, 7 and 13 corresponding to myeloid cells. (C) Clusters significantly affected by stimulation (left) with cluster histograms indicating co-secretion of IL-6. (D) Cluster analysis of IL-4 secreting with cluster 3, 7, and 11 corresponding to myeloid cells. (E) Clusters significantly affected by stimulation (left) and co-expression with IL-6 and IL-10 (right). (F) Comparison of PIM stimulation on donors with active TB (ATB), latent TB (LTB) and healthy controls (HC) in cluster 11. (G) Myeloid clusters secreting IL-6 (H) significantly affected by stimulation. (I) Cell surface phenotype of indicated cluster. (J) Differential effect of PIM stimulation on cluster 7 cells in ATB, LTB, and HC. (K) Myeloid clusters secreting IL-10 with (L) clusters significantly affected by stimulation (left panel) and histograms indicating IL-10 co-expression with IL-6. (M) Myeloid clusters secreting TNF-ɑ (N) significantly affected by stimulation (left) with co-expression of GM-CSF and IL-6 (right). (O) Myeloid clusters secreting GM-CSF. (P) Clusters differently affected by stimulation (left) with co-expression of TNF-α and IL-6 (right). Statistical differences between stimulations in (A) were evaluated by Friedman’s test with Dunnet’s posttest, while comparisons within multiple clusters (C, E, H, L, N, P) 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) (F, J) were compared using Kruskal-Wallis with Dunn’s posttest (n=5/group) *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001.