Fig. 2.

Gal-1 augments the frequency and immunosuppressive capacity of CD8⁺CD122⁺PD-1⁺ Tregs. (A–C) Spleen and axillary and inguinal lymph nodes from WT or Lgals1^−/− C57BL/6 (Left) or BALB/c mice (Right) were processed and CD8⁺CD122⁺PD1⁺ (A), CD8⁺CD25⁺Foxp3⁺ (B), and CD8⁺CD28⁻ (C) T cells were analyzed. (D) Schematic representation of immunosuppression assays. CD8⁺CD122⁺PD-1⁺ Tregs were isolated from the spleens of WT or Lgals1^−/− C57BL/6 mice and were further cocultured with total labeled splenocytes at different ratios (1:1, 1:2, 1:4). (E and F) Proliferation and division rates of CD4⁺ and CD8⁺ responders T cells. (G) Determination of IL-10 in supernatants of coculture assays at 96 h. (H) Frequency of IFN-γ⁺ T cells in splenocytes cocultured with CD8⁺CD122⁺PD1⁺ Tregs from WT or Lgals1^−/− mice detected by flow cytometry. Data presented are the mean ± SEM from three independent experiments. n = 5 to 6 mice per group. ANOVA, Bonferroni multiple comparison test, *P < 0.05 and ***P < 0.001.