Fig. 5: IFNγ, IL-4, and IL-10 are produced by distinct T cells in response to α-syn.

a–d After eliminating non-lymphocytes/monocytes and doublet cells by forward and side-scatter and dead cells by Live/Dead stain, cells were gated based on their cytokine expression and then cytokine-expressing cells were gated based on their CD3⁺ and CD56⁺ expression. CD3⁺ T cells were gated based on their CD4 and CD8 expression (CD8⁺, CD8⁺CD4⁺, CD4⁺, CD8⁻CD4⁻), and other populations based on CD3⁻CD56⁻ and other markers as indicated; CD14⁻CD19⁺ (B cells), CD14⁺CD19⁻ (Monocytes), CD14⁻CD19⁻ (other) (Supplementary Fig. 5), and percentage live cells expressing cytokine from each of these were plotted. Samples with a frequency of cytokines below 0.02% of live lymphocytes after removal of background were excluded from this analysis. a IFNγ (n = 13), b IL-4 (n = 9), and c IL-10 (n = 6). Median ± interquartile range is indicated. d Boolean gating for combinations of cytokines shown as percentage of live cells, n = 20. Median ± interquartile range is indicated. The gating strategy is shown in Supplementary Fig. 5. e Plots were gated on IL-10⁺CD4⁺ T cells following negative control (DMSO) or α-syn stimulation. The gating strategy leading up to this step is shown in Supplementary Fig. 5. IL-10⁺ cells were then gated further on CD25 and CD127 expression (top row). Plots were gated on CD25⁺CD127^loCD4⁺ T cells and then on IL-10 production (bottom row).