Fig. 4: Ag⁺ B cell landscape.

A Dot plot shows the total percentage and the absolute number of CD19⁺ B cells. The central bar represents the mean ± SD. Kruskal–Wallis test (one-sided) with Benjamini–Hochberg correction for multiple comparisons. B Dot plot shows the percentage and absolute number of antigen-specific CD19⁺ B cells. The central bar represents the mean ± SD. Kruskal–Wallis test (one-sided) with Benjamini–Hochberg correction for multiple comparisons was used to test the differences among the nine groups. C UMAP plot shows the 2D spatial distribution of 25.866 antigen-specific B cells from healthy controls (HD) and patients with Multiple Sclerosis embedded with FlowSOM clusters. Heatmap of the median marker intensities of the 10 lineage markers across the 11 cell populations obtained with FlowSOM algorithm after the manual metacluster merging. The colors of cluster_id column on the left correspond to the colors used to label the UMAP plot clusters. Each color in the heatmap is referred to the median of the arcsinh marker expression (0–1 scaled) calculated over cells from samples. Blue represents lower expression. while red represents higher expression. Light gray histogram bar and values indicate the relative sizes of the clusters. Naive; TrB. transitional B cells; MBC Usw. memory B cell unswitched; MBC memory B cell, PB plasmablasts, atBC atypical B cell. D (Left) UMAP graphs stratified by therapy. (Right) Dot plot showing the percentages and absolute numbers of naïve and MBC IgA B cells. The central bar represents the mean ± SD. Kruskal–Wallis test (one-sided) with Benjamini–Hochberg correction for multiple comparisons was used to test the differences among the nine groups. E Anti-spike and anti-RBD IgG concentrations in plasma samples from HD and MS treated groups. The central bar represents the mean ± SD. Kruskal–Wallis test (one-sided) with Benjamini–Hochberg correction for multiple comparisons. Adjusted P-values are indicated in the figure. Plots A–E HD healthy controls (N = 13); Cladribine (N = 6); DMF: Dimethyl Fumarate (N = 14); DMF Lymphopenic: Dimethyl Fumarate Lymphopenic (N = 9); Fingolimod (N = 12); IFN: Interferon 1β (N = 13); Natalizumab (N = 15); Teriflunomide (N = 8); Rituximab/Ocrelizumab (N = 11).