Fig. 4.
Signature Th cells are enriched in medullary regions of MG thymi and rebound in the blood after thymectomy. a Heatmap of surface markers and cytokine expression profiles of FlowSOM-generated Th subsets. b Violin plots comparing the frequencies of TFH cells and the corresponding frequency of IL-21 expression among TFH cells in MG patients vs non-MG patients. c Violin plot showing the frequency of GM-CSF expressing Th cells (ThGM) and ThCD103 cells in the thymi of MG and non-MG patients. d Immunofluorescence labeling of medullary thymic regions showing GM-CSF (red), CD4 (green), CD3 (magenta) and DAPI (blue). Samples from 7 MG patients and 5 non-MG controls were analyzed. Scale bar: 30 µm. Images of the single labels are enlargements of specified regions. Representative images of two independent experiments (1 slide with 3 sections/patient each) are shown. Left: MG, early onset, female MG patient without immunosuppressive therapy and thymus follicular hyperplasia, right: CTRL, patient with incidental mass, residual thymus tissue. HC Hassall’s corpuscle. e Heatmap of surface markers and cytokine expression profiles of FlowSOM-generated Th subsets in blood and thymus (left panel). Thymic expression profiles from both panels were clustered analogously using FlowSOM and displayed as combined expression profiles. Scaffold of the blood and thymic Th cell compartment in MG patients (right panel). Color overlay depicts expression of CD103 and CD69. f Heatmap of expression profiles of FlowSOM-generated cell clusters for quantitative immunofluorescence data of thymic regions after cell segmentation (middle panel). Colors correspond to the populations shown in Fig. S5f. Violin plots showing the frequency of ThCD103 cells within DAPI+ cells of MG patients and incidental mass lesion controls (right panel). PANCK pan-Cytokeratin. g Violin plots showing the frequency of ThGM and ThCD103 cells, and the frequency of IL-22 production within ThCD103 cells, in the blood of MG patients with or without thymectomy that did not receive further immunomodulatory treatment. h Violin plots showing the frequency of ThCD103 and ThGM cells in the blood of MG patients without treatment or with thymectomy (TMC) and/or azathioprine (AZA) treatment. Violin plots contain a bold horizontal line depicting the respective group mean. If not indicated, differences between experimental groups were statistically not significant (p > 0.05) using a nonparametric Mann–Whitney–Wilcoxon test with a false-discovery correction according to the Benjamini–Hochberg approach. *p < 0.05; **p < 0.01