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. 2019 Dec 19;9:19471. doi: 10.1038/s41598-019-55852-x

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© The Author(s) 2019

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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Differential abundances of lymphoid cell subsets in patients with early MS. (a) t-SNE map highlights mixed T and NK cell population (red dots). Heat map and cluster analysis of the mixed CD4⁺ T cell and NK cell population from all samples on the basis of the mean marker expressions (Panel B). Clusters are indicated by dendrograms. Heat colours show marker expression levels (red, high expression; dark blue, no expression). The bar graph shows mean cluster frequencies as % of parent (black number) and % of total cells (grey number). Cluster 3 and 6 (* in red) show differential abundances between healthy controls and patients with early MS. (b) The graph shows differences in frequency (%) of cluster 3 and 6 between the two studied groups (CON, grey; early MS, red). (c) Median expression levels as shown in the dot plot representation for randomly selected cells (n = 256/cluster) in the cluster 3 (orange), 6 (green) and all other clusters (blue). IL-6 was defined as a marker discriminating the differentially abundant cluster 6. (d) The representative t-SNE plot shows IL-6 expression levels and indicates the IL-6-expressing cell subset (orange circle, left image). The right image shows expression levels of IL-6 expressing cluster 6 (orange line), compared to the total cells (blue line). (e) t-SNE map highlights HLA-DR⁺ lymphocyte population (light blue dots). Heat map and cluster analysis of B cells from all samples on the basis of the mean marker expressions (Panel B). Cluster 7 (* in red) show differential abundances between the studied groups. (f) The graph shows differences in frequency (%) of cluster 7 between the two studied groups (CON, grey; early MS, red). (g) Median expression levels as shown in the dot plot representation for randomly selected cells (n = 256/cluster) in the cluster 7 (orange) and all other clusters (blue). MIPβ was identified as a marker discriminating the differentially abundant cluster 7. A P value < 0.05 at 10% FDR was considered statistically significant, determined using GLMM (**P < 0.01, unadjusted).