Fig. 2: Kinetics and phenotypic analysis of S-specific B-cells in kidney disease patients and controls.

a A pseudo-colored B-cell density plot (from red to blue, highest to the lowest density of events) of multicolor flow cytometry data after dimensionality reduction by t-distributed stochastic neighbor embedding (t-SNE) from concatenated, normalized, live, single CD19+ events from all individuals and timepoints (left panel) based on the expression of the 10 indicated markers as determined by the mean fluorescent intensity (MFI, right panels). Four dominant regions (Region 1-4) are outlined. b Mean and standard deviation of the frequency of events over the 4 regions per study group. Significant differences determined using two-way ANOVA and Tukey’s multiple comparison test are indicated (***p < 0.005 and ****p < 0.0001). c Longitudinal frequencies of live single S-specific CD19+ B-cells identified by classical gating in controls, CKD, dialysis and KTR pre-, 28 days and 6 months post-vaccination. Significant differences determined by Wilcoxon test for paired data are indicated (*p < 0.05). d) S-specific B-cells were overlayed on the t-SNE plot and depicted separately per study group and color-coded timepoint (red, blue and green).