Fig. 3: Phenotypic analysis of CD4⁺ Tconv cell subsets, CD4⁺ Treg cell subsets and CD19⁺ B cells in HDs, ECs, and non-ECs.

Comparative immune phenotypic analysis of a CD4+Tconv and CD19+B cells and b CD4+Treg cells in non-ECs, ECs, and HDs. aI SPADE tree with the distribution of CD4⁺Tconv subsets in HDs (black), ECs (green), and non-ECs (red). Nodes are colored by count. CD4⁺Tconv cells can be classified into four major subsets by their expression of CD45RA and the chemokine receptor CCR7: naïve (CCR7⁺CD45RA⁺), CM (CCR7⁺CD45RA⁻), EM (CCR7⁻CD45RA⁻), and TEMRA (CCR7⁻CD45RA⁺). aII Frequency of naïve, CM, EM, and TEMRA cells in each studied group (HDs n = 24, ECs n = 16, and non-ECs n = 23). Boxplots show the expression of the indicated marker in CD4⁺ CM cells (aIII) across the groups (HDs n = 22, ECs n = 12, and non-ECs n = 8). aIV Radar chart of the composite scores of phenotypic cell alteration calculated for each CD4⁺Tconv subpopulation in non-ECs and ECs (see Methods). aV Frequency of circulating T follicular helper cells (cTfh), expression levels of CXCR5 in cTfh, and frequency of cTfh co-expressing CD38 and HLA-DR in non-ECs (n = 6) and HDs (n = 5). aVI Proportion of CD19⁺ B cells and frequency and expression level of CXCR5 in CD19⁺B cells in non-ECs (n = 6) and HDs (n = 5). bI Representative flow cytometry plots of CD25⁺Foxp3⁺ cells within CD4⁺T cells isolated from HDs (n = 22), ECs (n = 12), and non-ECs (n = 8). bII Histograms with the frequency of Foxp3 in CD4⁺T cells and bIII displaying the CD25 expression level in CD4⁺ Foxp3⁺T cells and bIV the regulatory T cell (Treg) CD25⁻ variant frequency in CD4⁺Foxp3 T cells in each studied group. bV Scatterplots of the relationships between frequency of the Treg CD25⁻ variant in HIV-infected patients and serum IFNα levels. bVI Proportion of a specific functional signaling checkpoint on memory CD4⁺ Treg (CD4⁺ Foxp3⁺CD25⁺CD45RA⁻) cells of each studied group (HDs n = 22, ECs n = 12, and non-ECs n = 8). Correlations were evaluated using Spearman’s rank correlation test. Comparisons between the two groups were made with the Mann–Whitney U-test. Multiple group comparisons were assessed using the Kruskal–Wallis test with Dunn’s multiple comparison testing. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001. Error bars on graphs represent interquartile ranges.