(A) Schematic for adoptive transfer of “Acute-7d” (blue) or “T_Dysf” (red) P14 cells into congenically distinct C57BL/6 mice on day 19, followed by acute LCMV infection and analysis on day 6–7 postinfection. (B) Total P14 cell numbers from spleens, livers, and lungs. Panels C–G and I–N are for spleens. (C) Bar graph showing expression level (gMFI) of Ly108 or (D) % Ifnγ⁺ P14 cells after ex vivo GP33 peptide rechallenge. (E) Representative FACS plots showing Ifnγ and Perforin expression and summary bar graph showing % Ifnγ⁺Perforin⁺ P14 cells. (F) Bar graph showing expression level (gMFI) of Gzmb or (G) TOX. (H) Schematic for adoptive transfer of “Rested Acute-7d” (dark blue) or “Rested T_Dysf” (dark red) P14 cells on day 26 into congenically distinct C57BL/6 mice, followed by acute LCMV infection and analysis on day 6–7 postinfection. (I) Bar graph showing total number of P14 cells isolated from spleens or (J) expression level (gMFI) of Ly108. (K) Representative FACS plots showing Ifnγ and Tnf or CD107a expression and summary bar graph showing % of Ifnγ⁺Tnf⁺ or Ifnγ⁺CD107a⁺ P14 cells. (L) Representative FACS plots showing PD-1 and Tim3 expression and (M) summary bar graph showing % of PD-1⁺Tim3⁺ P14 cells. (N) Bar graph showing the expression level (gMFI) of Gzmb. For all FACS plots (E, K, and L), Acute-7d plots are representative of 1 recipient mouse, while the T_Dysf plots show data combined from all T_Dysf recipient mice. For B, data were pooled from 2 independent experiments with n = 3–5 biological replicates per group for each experiment. For all other panels, n = 4–6 biological replicates from 2 to 3 independent experiments. Adjusted P value *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001. Comparisons were determined by Mann-Whitney U test (unpaired, 2-sided). Error bars indicate mean ± SEM.