Expression of cell cycle–associated genes and division are sustained in CD8 T cells when activated in the presence of inflammation. Mice were injected with OT-I CD8 T cells and immunized with DC or DC+CpG as in Fig. 1. BrdU was injected in an ∼15-h pulse. The frequency (A and C) and absolute number (B and D) of OT-I CD8 T cells (A and B) or OVA257-264 tetramer+ CD8 T cells (C and D) in spleen was measured by flow cytometry. Data are represented as mean ± SEM. Statistical analysis used Student’s t test (n = 3 from each group). **, P < 0.01; NS = P > 0.05. Data are representative of two independent experiments. (E–H) OT-I T cells were isolated from the spleen at day 5 (E and H) or day 7 (F–H) after immunization, and mRNA levels of Ccna2, Ccnb1, Ccnb2, Ccne2, Foxm1 (E and F) were analyzed by RT-PCR and the indicated FoxM1 target genes (G) were analyzed using microarray data previously described in Fig. 1. mRNA levels in (E and F) are expressed relative to naive OT-I T cells; mRNA levels in G are displayed as fold expression in cells from DC+CpG- versus DC alone-immunized mice. Protein expression of Cyclin A, Cyclin B1, and FoxM1 was analyzed by Western blot (H). Data in E and F are represented as mean ± SEM. Statistical analysis used Student’s t test (n = 3 from each group). **, P < 0.01; NS = P > 0.05. Data in E, F, and H are representative of two independent experiments. *, FDR < 0.01.
