Figure 1. High-Dimensional Analyses of Activated T Cells in HIV-Infected LNs.

(A–E) CyTOF analyses of 8 HIV-infected LNs.

(A) Representative plot and bar graph show the relative contribution of major T cell subsets in HIV-infected LNs by TCR and co-receptor expression.

(B) Plot summarizes fold change over mean signal intensity for differentially expressed markers between CD38⁻HLA-DR⁻ and CD38⁺HLA-DR⁺ CD3⁺ T cells.

(C) UMAP displays of PD-1 and CXCR5 staining on CD4⁺ T cells using concatenated data from 8 HIV LN samples.

(D) Heatmap shows the average staining signal of individual markers for each CD4⁺ T cell subset as indicated. Markers used to select input cells were excluded.

(E) Representative plot showing PD-1 and CD38 co-staining.

(F–J) Flow cytometry analyses of 7 HC and 9 HIV LNs.

(F) Plots showing subdivision of CD4⁺ T cells by CXCR5 and PD-1 staining on a representative HC or HIV-infected LN sample.

(G) Bar graph quantifies the frequency of each phenotypic subset in HC or HIV-infected LNs.

(H) The frequency of CD38⁺ T cells within each manually gated CD4⁺ T cell subset.

(I) The frequency of CD38⁺ T cells within ICOS⁻ versus ICOS⁺ subset of CXCR5⁻PD-1⁺⁺ T cells.

(J) Correlation between peripheral CD4:CD8 ratio of 22 HIV⁺ donors and the frequency of CXCR5⁻PD-1⁺ICOS⁺ subset of CD4⁺ T cells in their LNs.

For (B) and (G), differentially expressed markers were selected using multiple t tests and corrected using Holm-Sidak method. For (H), Friedman test was performed and corrected using Dunnett’s multiple comparisons test. For (I), paired t test was used. Association for (J) was measured by Pearson correlation. Data are represented as mean ± SEM.

Also see Figures S1 and S2.