Reconstitution of CD8⁺ DCs in LNs inhibits antigen-specific T-cell responses. Mice were treated with CTX on day −4, followed by the adoptive transfer of 10⁵ CD8⁺ pmel-1 cells from Thy1.1 pmel-1 mice by intravenous injection on day −1. On day 0, mice were immunized with hgp100 peptide–pulsed DCs injected intradermally in the left ear. At 5 days after immunization, draining retroauricular LNs were harvested, and stained for CD8 and Thy1.1 to detect antigen-specific pmel-1 cells (Thy1.1⁺). To assess IFN-γ production, cells were cocultured with mgp100-pulsed EL-4 cells and then stained for intracellular IFN-γ. All mice received adoptively transferred pmel-1 CD8 T cells. As negative control for immunization, mice were not immunized and treated (▴) or not treated (■) with CTX. As negative control for CTX, immunized mice did (▾) or did not (♦) receive CTX treatment. A group of mice received 10⁶ CD8⁺ DCs in the left ear (●) or received 10⁶ CD8⁻ DCs in the left ear (□) on day −1. (A) Experiment timeline. (B) Representative plots. (C) The recovery of antigen-specific T cells and percentage of IFN-γ production presented as the mean ± SEM (3-4 mice/group). Data are representative of 3 independent experiments. CD8 pmel-1 recovery and IFN-γ secretion were both statistically significantly enhanced when CTX (♦) or CTX and CD8-DC (□) treatment preceded immunization (P = .05).