Figure 2.

Enhancement of CD8⁺ effector function by castration was diminished 5 weeks after immunization. A, the representative dot plot (left) shows PE- and Cy5.5-tetramer double-positive cells gated on the CD8⁺CD44⁺ population as described in Fig. 1. The number of tetramer-positive cells was quantified and represents the average of 5 animals in one experiment (bar graphs; ■, Cx + Immun; □, Sx + Immun). B, IFN-γ–secreting cells in response to ex vivo UV-8101-RE stimulation were identified using an ELISPOT assay with PDLNs (left) and spleen cells (right) from both groups. The average of triplicate wells in one representative experiment is shown. C, lytic capacity of MLTC cells cultured for 6 days was determined by ⁵¹Cr release assay (◆, UV-8101-RE; ▲, TRAMP-C1). Line graphs show one representative animal from each group and the bar graph shows the average of all animals in each group at an E:T ratio of 50:1. D, IFN-γ expression by MLTC cells was assessed by an intracellular cytokine staining assay (left). Percentages of IFN-γ–expressing cells gated on CD8⁺ T cells were calculated (right). Data are presented as mean values ± SD and one representative experiment of 3 independent experiments with 3 to 5 mice per group is shown. Cx, castration; Immun, immunization; Sx, sham treatment.