Attenuation of cTfh expansion and dysregulation on sirolimus treatment. (A) Representative flow plots showing frequency of cTfh cells in a patient with m-IC before (Pre) and after (Post) sirolimus treatment. (B-C) Frequency of PD-1⁺ cTfh (PD-1⁺ CXCR5⁺) and total cTfh (CXCR5⁺) are shown for HC and patients with m-IC (n = 9) before and after sirolimus therapy. (D-F) Plots showing frequencies of cTfh activation (HLA-DR⁺ CXCR5⁺), exhaustion (Tim3⁺ CXCR5⁺) and senescence (CD57⁺ CXCR5⁺) gated on total memory CD4⁺T (CD45RA^-CD4⁺) cells. (G-H) cTfh population was gated on CD4⁺ naïve T and CD4⁺ TEMRA T cells and plots showing percentage of these populations pre- and post-sirolimus therapy. (I) Violin plot showing percentage of cTfh in naïve, memory and TEMRA compartments of CD4⁺ T -cells in HC and patients with m-IC before and after sirolimus treatment. Kruskal-Wallis 1-way ANOVA followed by Dunn’s multiple comparison test for non-normally distributed samples and ordinary 1-way ANOVA followed by Tukey’s multiple comparison test for normally distributed samples were used for statistical comparison of HC with m-IC pre- and posttreatment groups. Paired t test or Wilcoxon signed-rank test was used for paired analysis between pre- and post-m-IC groups. ∗P < .05, ∗∗P < .01, ∗∗∗P < .001, ∗∗∗∗P < .0001; ns, not significant.