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. 2016 Mar 28;90(8):3913–3928. doi: 10.1128/JVI.02450-15

FIG 4.

FIG 4

More exhausted HSV-specific CD8+ T cells were detected in acutely HSV-1 LAT+ infected TG than in HSV-1 LAT– infected TG. Three groups of five HLA Tg rabbits were ocularly infected as described above with the wild-type McKrae (LAT+), its LAT-null virus mutant dLAT2903 (LAT–), or the marker-rescued virus dLAT2903R counterpart (LAT+), as in Fig. 2. Ten TG/group were harvested at 12 days postinfection, pooled, and processed, as described above. The cells were triple stained with FITC-labeled anti-PD-1, PerCP-anti-CD8 T-cell, and PE-labeled HSV-1 VP11/12220–228/HLA Tg rabbit-A*0201 tetramer. The levels of PD-1 and TIM-3 expression and the percentages of HSV-1 VP11/12220–228 epitope-specific CD8+ T cells expressing PD-1, TIM-3, and CTLA-4 were determined by FACS. (A) Representative histogram of levels of PD-1 on gated VP11/12220–228 epitope-specific CD8+ T cells in LAT+ TG versus LAT– TG. (B) Representative dot plots of the percentages of VP11/12220–228 epitope-specific PD-1+ CD8+ T cells in LAT+ TG versus LAT– TG. (C) Representative histograms of the levels of TIM-3 gated on VP11/12220–228 epitope-specific CD8+ T cells in LAT+ TG versus LAT– TG. (D) Representative dot plot of the percentages of VP11/12220–228 epitope-specific TIM-3 CD8+ T cells in LAT+ TG versus LAT– TG. (E) Percentage of VP11/12220–228 epitope-specific CD8+ T cells expressing PD-1. (F) Percentage of VP11/12220–228 epitope-specific CD8+ T cells expressing TIM-3. (G) Percentage of VP11/12220–228 epitope-specific CD8+ T cells expressing CTLA-4. Each bar in panels C and F represents the mean and SD from two independent experiments. *, P < 0.05.