Fig. 2.

Gal-9 knockout animals mount stronger virus-specific CD8 T-cell responses in the acute phase. Virus-specific CD8 T-cell responses were compared among age- and sex-matched IAV-infected WT and G9KO animals at indicated time points p.i. Representative FACS plots show NPtet⁺CD8 T cells from WT and G9KO animals in the BAL (A) and spleen (B). (C) Absolute numbers of NPtet⁺ and IFNγ⁺TNFα⁺ CD8 T cells at day 10 p.i. in BAL. Bar diagram shows the frequencies (D) and absolute numbers (E) of NPtet⁺ CD8 T cells in spleen. (F) MFI of cytokine IFNγ produced by CD8 T cells in BAL. (G) Histograms showing Tim-3 expression on total BAL CD8 T cells in WT (light line) and G9KO mice (darker line) at day 10 p.i. Representative FACS plots show the frequencies of influenza A peptide NP_366–374 (ASNENMETM) stimulated polyfunctional (IFNγ⁺TNFα⁺) CD8 T cells isolated from the BAL (H, Upper) and spleen (H, Lower) at day 10 p.i. of WT and G9KO animals. (I) Tim-3 expression by IFNγ⁺CD8 T cells in WT and G9KO animals at day 10 p.i. following stimulation with NP_366–374 peptide. (J) Absolute numbers of Tim-3⁺, IFNγ⁺, IFNγ⁺TNFα⁺ CD8 T cells in the spleen at indicated time points p.i. Coexpression of Tim-3 and NPtet⁺ with CD44 in BAL (K), spleen (L), and with CD62L in BAL (M) and spleen (N) at day 8 p.i. is shown in WT and G9KO animals by representative FACS plots. Data are representative of three independent experiments with three mice per group in each experiment. Error bars represent SEM.