Fig. 2.

Reduced cytokine polarization and systemic TNF-producing CD103⁺ T cells represent an MG-specific signature. a Heat map showing relative expression of the indicated activation markers within naïve, memory and effector subsets of CD4⁺ T cells obtained by FlowSOM clustering. b Radar plot (left panel) representing the cytokine profile of antigen-experienced CD4⁺ T cell subsets (T_Eff, T_EM and T_CM). The colored line indicates the Cohen’s d effect size for each cytokine (MG vs. CTRL) as a deviation from the gray dashed reference line. Th cytokine profiles were manually annotated based on partially overlapping key cytokines. The violin plot (right panel) shows the frequency of GM-CSF⁺ cells among antigen-experienced CD4⁺ T cells in control and MG patients. T_CM central memory T, T_EM effector memory T, T_Eff effector T, T_N naive T. c Subset composition of the GM-CSF-producing Th cell population within all patients’ blood (MG and CTRL; upper panel); and relative abundance of GM-CSF-producing cells within the CD4⁺ T_EM population (bottom panel) in control and MG patients (left), and in low and high disease severity newly diagnosed treatment-naïve MG patients (right). Clinical disease severity was determined by the modified quantitative MG score, with low disease severity scoring < 0.5, and high disease severity ≥ 0.5. d Cytokine expression analysis of CD4⁺ T_EM cells. FlowSOM yielded 13 cytokine expressing clusters (c1–c13; determined by consensus clustering). Corresponding expression profiles (left box) as well as statistical parameters (right box) are displayed. Blue color indicates high significance (low p value) for the comparison of treatment-naïve MG patients vs. CTRL, red color indicates high significance and high R² value, respectively, for the correlation with the continuous clinical disease severity. e Heatmap comparing follicular Th cells (T_FH) and CD103⁺ Th cells (Th_CD103) to conventional Th cells. f Row-normalized heatmap of cytokine positivity for 13 detected cytokines among peripheral Th subsets for all patients present in the cohort. g Correlation between the frequency of TNF-producing Th_CD103 cells (left panel) or serum anti-AChR antibody titer (right panel) and the modified quantitative MG score as a measure of clinical disease severity for newly diagnosed patients neither receiving immunomodulatory nor symptomatic 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. For correlation analysis, statistical parameters were obtained using a linear regression model. Shaded areas in g represent the 95% confidence interval