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

Fig. 1.

Fig. 1

Heterogeneity of SLAMF7 expressing T cells. (A) Heterogeneity of T cells by high-dimensional single-cell flow cytometry staining. Frozen PBMCs were thawed and stained for flow cytometry. T cells of 20 donors (n = 10 each for young and older donors, all CMV seropositive) were concatenated. T cells were visualized by UMAP and a cold to hot heat map was used to represent the intensity of each marker. (B) Characterization of human CD8 T cell maturation. The mRNA fold change expression in comparison to naïve CD8 T cells was calculated in sorted populations of T Stem Cell Memory (TSCM), Central Memory (TCM), and Effector Memory (TEM), top. The mRNA coding for transcription factors is labelled in red and surface molecules in black. The cellular progression according to the diffusion map was visualized by the expression profile of CD45RO, SLAMF7, CX3CR1, KLRG1, GPR56, CD27, CD57, T-bet, RunX3, and HOBIT in CMV seropositive donors, bottom. (C) SLAMF7 expression in CD8 T cell subsets during aging. The frequency of SLAMF7 was evaluated in naïve (CD45RO-CCR7+CD27+CD95-), TSCM (CD45RO-CCR7+CD27+CD95+), TCM (CD45RO+CCR7+CD27+CD95+), TTM (CD45RO+CCR7-CD27+CD95+), TEM (CD45RO+CCR7-CD27-CD95+), and TTE (CD45RO-CCR7-CD27-CD95+) of frozen PBMCs from young and older donors (n = 10 and n = 11 respectively). The statistical analysis was performed on unpaired samples (U Mann–Whitney test) (** and **** for p < 0.01 and p < 0.0001, respectively). (D) Heterogeneity of SLAMF7 expressing T cells during aging. Staining was performed on frozen PBMCs of young (n = 10) and elderly donors (n = 10). The statistical analysis was performed on unpaired samples (U Mann–Whitney test; *, **, ***, and **** for p < 0.05, p < 0.01, p < 0.001, and p < 0.0001, respectively). (E) SLAMF7 expression in CD8 T cell subsets during untreated HIV infection. The median intensity of SLAMF7 expression was evaluated in subsets (as defined in Fig. 1C) of frozen PBMCs from PLWH and healthy donors (n = 15 and n = 12 respectively). The statistical analysis was performed on unpaired samples (U Mann–Whitney test) (** and **** for p < 0.01 and p < 0.0001, respectively). (F) SLAMF7 expression in CD8 T cell subsets during chronic viral infections and aging. The frequencies of SLAMF7 expression were evaluated in total CD8 T cells from frozen PBMCs of PLWH under anti-retroviral therapy and co-infected or not with CMV. Patients have been stratified by age (< 40, 40–65, > 65 years old as young, middle age, and old with n = 42, n = 43, and n = 11 respectively) and compared to healthy donors (n = 20). The statistical analysis was performed on unpaired samples (U Mann–Whitney test) (** and **** for p < 0.01 and p < 0.0001, respectively). (G) Identification of progenitor-like and terminal effector CD8 T cells based on SLAMF7 expression. Description of the gating strategy used to identify SLAMF7+-progenitor-like (CD27+GPR56-) and terminal effector (CD27-GPR56+) population in total CD8 T cells. The histograms represented the overlaid intensity of TCF-1, CD127, SLAMF6, and CD5 in SLAMF7 subsets in comparison to Naïve/ Central Memory (CD27+CCR7+) CD8 T cells. (H) Characterization of progenitor-like and terminal effector CD8 T cells. The median intensity of fluorescence was compared between progenitor-like and terminal effector CD8 T cell populations. The statistical analysis was performed on paired samples (n = 20, Wilcoxon signed-rank test), (** and **** for p < 0.01 and p < 0.0001, respectively).