Poor polyfunctionality in KI mice is the result of defective responses to TCR restimulation. (A) Splenocytes from day 8 postinfection WT, Y145F, and Y112/128F mice were stimulated with PMA plus ionomycin. Representative contour plots show IFNγ and TNFα (left panel) and IFNγ and IL-2 (right panel) expression in CD8⁺ cells. Numbers indicate the percentage of CD8⁺, IFNγ, TNFα (left panel), or IFNγ, IL-2 (right panel) double producers over the percentage of total CD8⁺ IFNγ producers. (B) Dot plots show IL-7rα and KLRG-1 surface expression on H2D^b:GP33-reactive CD8⁺ PBLs from WT, Y145F, and Y112/128F mice 15 days postinfection (left panel). Numbers indicate the percentage of CD8⁺ H2D^b:GP33-reactive lymphocytes within each gate. Top left and bottom right gates from top panel were used to quantify the percentage of SLECs (middle panel) and MPECs (right panel), respectively. For graphs each point represents an individual mouse; bars represent average ± SEM. Data are representative of 3 independent experiments with 5-10 mice per group. Significant P values, when present, comparing KI to WT are indicated by asterisks: ***P < .001, **P = .001-.01, *P = .01-.05.