Figure 7.

Identification of a rare population of CXCR5⁺CD8⁺ central memory T cells that is not activated by QIV. (a) Gating strategy and t-SNE of live CD3⁺CD8⁺ lymphocytes of all participants at all time points. Heatmaps show the relative expression of each marker. (b) t-SNE of live CD3⁺CD8⁺ lymphocytes of one C-HIV participant showing the CXCR5⁺ and CXCR5⁻populations. (c) Characterisation of CD8⁺CXCR5⁻(blue peaks) and CD8⁺CXCR5⁺ (red peaks) T cells using histograms derived from t-SNE on concatenated data from all individuals to determine the relative expression of cellular markers. (d) FACS gating strategy for CD8⁺CXCR5⁺ T cells and T cell memory subsets. (e) Comparison of the frequency of CXCR5⁺CD8⁺ T cells in HC, E-HIV, C-HIV at all time-points (f) Comparison of the frequency of CD8⁺CXCR5⁺ (red columns) and CD8+CXCR5^- (blue columns) T cells that were, from left to right, central memory cells (T_CM: CD45RA⁻CCR7⁺), (T_EM: CD45RA⁻CCR7⁻), (Naive: CD45RA⁺CCR7⁺), (T_EMRA: CD45RA⁺CCR7⁻). Comparison of the frequency of CD8⁺CXCR5⁺ and CD8⁺CXCR5⁻T_CM-cells expressing (g) PD-1 and (h) CD32. (i) Frequency of CD8⁺CXCR5⁺ PD-1⁺ T cells and (j) CD8⁺CXCR5⁻PD-1⁺ T cells at all time⁻points. P values were derived using a Kruskal-Wallis or Friedman test (p value below graph) and p values of the Dunn’s multiple comparisons test (above graph). Comparisons between groups were performed using a two-tailed Mann-Whitney U test.