Figure 9. IL-10 production by human... | Rockefeller University Press

Figure 9.

$IL-10 production by human influenza-specific tissue memory T cells is associated with enhanced effector function. (A) Schematic illustrating the samples obtained from organ donors (n = 6, D), living subjects (n = 6, FLD), and blood and tissue samples profiled. (B) Plots show identification of influenza (flu)-reactive CD4+ (orange) and CD8+ (green) T cells using the AIM assay following stimulation with peptide pools. (C) Representative flow cytometry plots of unstimulated (top) and flu peptide pool–stimulated (bottom) CD4+ (orange, left) and CD8+ (green, right) T cells. (D and E) Compiled unstimulated and flu-reactive T cells visualized by UMAP colored by cell lineage defined by a multimodal classifier (see Materials and methods), stimulation condition, tissue, and donor (E) GSEA enrichment of top 500 mouse secondary response genes within the human flu-reactive T cell response. Gene sets were selected from all differentially expressed genes or those from the T cell activation cluster (Fig 1 D). (F) Heatmap depicting the row-normalized expression of T cell–associated cytokines enriched in mouse flu secondary response (identified in Fig. 2) in resting and flu-reactive T cells across human circulation (blood and spleen) and tissue (lung and LLN). * indicates genes that are significantly upregulated in the mouse or human flu response. (G–I) Bar plots depicting the mean IL10 log-normalized expression across (G) tissue and stimulation conditions in all cells evaluated, (H) within flu-reactive CD4+ and CD8+ T cells, and (I) classifier-defined T cell phenotypic subsets. Statistical significance for G–I was determined by one-way ANOVA corrected for multiple comparisons with Tukey’s HSD test. (J and K) Expression of residency and circulatory (J) genes and (K) surface markers on flu-reactive T cells in tissue. Dot plots depict gene expression: the dot size represents the fraction of cells in that group expressing a gene, and the dot color represents the row-normalized expression. Violin plots display the distribution of ADT expression for each population colored by the median log-normalized ADT counts per thousand. (L) Scatterplot showing average R values vs log-normalized gene expression. R values were calculated for each protein-coding gene expressed in >80 cells, and plotted R values are for those genes with the same sign across all donors evaluated (see Materials and methods). (M) Dot plot depicting gene expression of AIM and functional mediators in unstimulated or flu-reactive, IL10-positive or IL10-negative cells. The dot size represents the fraction of cells in that group expressing a gene, and the dot color represents the row-normalized expression. *P < 0.0001. ADT, antibody-derived tag. Refer to the image caption for details.$

IL-10 production by human influenza-specific tissue memory T cells is associated with enhanced effector function. (A) Schematic illustrating the samples obtained from organ donors (n = 6, D), living subjects (n = 6, FLD), and blood and tissue samples profiled. (B) Plots show identification of influenza (flu)-reactive CD4⁺ (orange) and CD8⁺ (green) T cells using the AIM assay following stimulation with peptide pools. (C) Representative flow cytometry plots of unstimulated (top) and flu peptide pool–stimulated (bottom) CD4⁺ (orange, left) and CD8⁺ (green, right) T cells. (D and E) Compiled unstimulated and flu-reactive T cells visualized by UMAP colored by cell lineage defined by a multimodal classifier (see Materials and methods), stimulation condition, tissue, and donor (E) GSEA enrichment of top 500 mouse secondary response genes within the human flu-reactive T cell response. Gene sets were selected from all differentially expressed genes or those from the T cell activation cluster (Fig 1 D). (F) Heatmap depicting the row-normalized expression of T cell–associated cytokines enriched in mouse flu secondary response (identified in Fig. 2) in resting and flu-reactive T cells across human circulation (blood and spleen) and tissue (lung and LLN). * indicates genes that are significantly upregulated in the mouse or human flu response. (G–I) Bar plots depicting the mean IL10 log-normalized expression across (G) tissue and stimulation conditions in all cells evaluated, (H) within flu-reactive CD4⁺ and CD8⁺T cells, and (I) classifier-defined T cell phenotypic subsets. Statistical significance for G–I was determined by one-way ANOVA corrected for multiple comparisons with Tukey’s HSD test. (J and K) Expression of residency and circulatory (J) genes and (K) surface markers on flu-reactive T cells in tissue. Dot plots depict gene expression: the dot size represents the fraction of cells in that group expressing a gene, and the dot color represents the row-normalized expression. Violin plots display the distribution of ADT expression for each population colored by the median log-normalized ADT counts per thousand. (L) Scatterplot showing average R values vs log-normalized gene expression. R values were calculated for each protein-coding gene expressed in >80 cells, and plotted R values are for those genes with the same sign across all donors evaluated (see Materials and methods). (M) Dot plot depicting gene expression of AIM and functional mediators in unstimulated or flu-reactive, IL10-positive or IL10-negative cells. The dot size represents the fraction of cells in that group expressing a gene, and the dot color represents the row-normalized expression. *P < 0.0001. ADT, antibody-derived tag.