Figure 4.

Vaccinated mice challenged with CVB4 induce the generation of VP1-specific CD4 T cells. Lymphocytes were prepared from the indicated groups as described in the methods. Cells were stimulated with VP1 721–740 and maintained in IL-2 medium supplemented with antibiotics. Viable cells were harvested between 7 and 11 days post-stimulation and were stained with the indicated IA^k tetramers, anti-CD4, and 7-AAD. After cells were acquired by flow cytometry, tetramer⁺ cells were analyzed in the live (7-AAD¯) CD4⁺ subset using FlowJo software. RNase 43–56, control. Representative flow cytometric dot-plots and the mean ± SEM values from three individual experiments, each involving n = 3–6 mice, are indicated. Unpaired Student’s t-test (two-tailed) was used to determine significance between treatment groups. * p ≤ 0.05 and ** p ≤ 0.01.