Fig. 1.
The canonical peripheral immune landscape of MG patients does not differ from that of healthy individuals. a Cryopreserved peripheral blood mononuclear cells (PBMCs) from myasthenia gravis patients (MG, n = 38) and healthy controls (CTRL, n = 21) were labeled with a panel of antibodies recognizing either surface markers or intracellular cytokines (following brief antigen-independent restimulation) and data acquired by CyTOF. Thymic leukocytes from MG patients (n = 4) and non-MG incidental mass lesion controls (n = 6) were analyzed in a similar manner by partially overlapping spectral flow cytometry panels. Thymic tissue sections of MG patients (n = 13) and non-MG controls (n = 6) were analyzed by quantitative multiplexed immunofluorescence microscopy. The resulting datasets were analyzed using a data-driven high-dimensional approach using supervised and unsupervised machine-learning algorithms. b UMAP of 100,0000 cells randomly sampled from the combined dataset. Color code indicates FlowSOM clustering and manual annotation according to lineage marker expression profiles presented in the heatmap. Tregs regulatory T cells, DCs dendritic cells, NK natural killer. c Violin plots showing the frequency of the FlowSOM-generated immune clusters in healthy controls and MG patients that did not receive immunomodulatory treatment. d Violin plots showing the frequency of memory B cells (BMEM), plasmablasts (PB) and peripheral follicular T helper cells (TFH) in healthy controls and MG patients obtained by subclustering the B cell and CD4+ T cell compartments. Violin plots contain a bold horizontal line depicting the respective group mean. If not indicated, the 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