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. 2024 Dec 28;14:30779. doi: 10.1038/s41598-024-80971-5

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© The Author(s) 2024

Open Access This article is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License, which permits any non-commercial use, sharing, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if you modified the licensed material. You do not have permission under this licence to share adapted material derived from this article or parts of it. The images or other third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by-nc-nd/4.0/.

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Fig. 5 — Homeostatic regulation of SLAMF7⁺ CD8 T cells. (A) Distribution of IL-15/ TCR stimulated CD8 T cells. Total CD8 T cells from stimulated PBMCs (n = 12) were concatenated and visualized by UMAP. The phenotype was measured after 4 days of stimulation in the presence of anti-CD3 microbeads with and without IL-15 (10 ng/mL). TCR-stimulated and TCR/IL-15-stimulated CD8 T cells were overlaid on the UMAP representation in black and red respectively. (B) Phenotype of IL-15/ TCR stimulated CD8 T cells. UMAP visualized the phenotype of activated CD8 T cells as in (A). Clusters were automatically determined by phenograph. A cold-to-hot heatmap represented the scaled intensity of each marker. (C) SLAMF7-associated phenotype of IL-15/ TCR stimulated CD8 T cells. The fluorescence intensity expression of each marker was overlaid for both conditions. (D) Acquisition of SLAMF7 and inhibitory receptors in response to IL-15/ TCR stimulation. The frequency of total and SLAMF7 expressing CD8 T cells was measured after 4 days of in vitro stimulation. The statistical analysis was performed on paired samples (Wilcoxon signed-rank test) (n = 12 and *, **, *** for p < 0.05, p < 0.01 and p < 0.001 respectively). (E) SLAMF7 signaling and T cell proliferation. The proliferation was measured by the dilution of Cell Trace Violet (CTV) after 4 days of IL-15/ TCR stimulation with and without SLAMF7 ligation. The expression of CD27, TCF-1, and TOX were quantified on proliferating T cells, defined as CTV^low. The statistical analysis was performed on paired samples (Wilcoxon signed-rank test) (n = 8 and *, ** for p < 0.05, and p < 0.01 respectively). (F) Accumulation of SLAMF7⁺ CD8 T cells during aging. The correlations between the frequencies of CD8 T cell populations and the chronological age were evaluated by the Spearman test, left. The IMM Age was calculated according to the expression of 57 pre-determined genes detected by RNA-Seq and correlated to the frequencies of SLAMF7 expressing CD8 T cells in the ATTRACT cohort (n = 27, r = 0.42 and p < 0.05). (G) Accumulation of SLAMF7⁺ CD8 T cells during inflammation. The quantification of sCD14 was performed by Elisa to measure the systemic inflammation in the plasma of PLWH. The correlation between the frequency of SLAMF7⁺CD8 T cells and the concentration of sCD14 was evaluated by a Spearman test.