Figure 5.

Proteome analysis, proliferation, and apoptosis of virus-specific CD8⁺ T cells in PD-L1^−/− mice. CD8⁺ T cells were isolated from CD45.1 × TCR Tg mice and adoptively transferred into WT or PD-L1^−/− mice. One day later recipient animals were infected with FV. CD8⁺ CD45.1⁺ T cells were sorted from spleens of WT and PD-L1^−/− mice and lysated for proteome analysis at day 12 after FV infection. (A) Volcano plot showing the comparison of protein expression in CD8⁺ CD45.1⁺ T cells isolated from WT and PD-L1^−/− mice. Significantly regulated proteins are highlighted as black dots. (B) The list is showing significantly regulated proteins and the ratio of protein expression in PD-L1^−/− to WT for every protein. (C) C57BL/6, PD-1^−/−, and PD-L1^−/− mice were infected with FV and splenocytes were isolated at different time points after infection. Flow cytometry was used to detect the numbers of virus-specific Tetramer⁺ CD8⁺ T cells expressing the intracellular proliferation marker Ki67. (D) C57BL/6, PD-1^−/− and PD-L1^−/− mice were infected with FV and splenocytes were isolated at day 8 after infection. Flow cytometry was used to detect the expression of activated caspase 3 in the cytoplasm of the virus-specific Tetramer⁺ CD8⁺ T cells. C57BL/6 mice were infected with FV and at day 6 and 7 after infection they were treated with pan-caspase inhibitor Z-WAD-FMK. Flow cytometry was used to detect numbers of virus-specific Tetramer⁺ CD8⁺ T cells in spleen (E) and bone marrow (F) of 8 days infected mice. Each dot represents an individual mouse and the mean numbers and SD are indicated. Differences were analyzed by Benjamini-Hochberg corrected one-way ANOVA (A,B), one-way ANOVA with a Tukey post-test (C,D), or an unpaired t-test (E,F). Statistically significant differences between the groups are indicated (*p < 0.05, ***p < 0.0005).