FOXO3 Regulates CD8 T Cell Memory by T Cell-Intrinsic Mechanisms
Figure 10
Loss of FOXO3 in the T cell compartment enhances the quantity of memory CD8 T cells without decreasing their quality.
(A) At 180 days after LCMV infection, mononuclear cells from spleen, liver and lymph nodes of +/+ and FOXO3L mice were collected and stained with anti-CD8 and MHC-I tetramers. Bars represent data from 2 independent experiments with 4–7 mice/group/experiment; error bars represent the SEM and * indicates statistical significance at p<.05. (B) At 180 days PI, splenocytes were stained with anti-CD44, anti-CD62L, anti-CCR7 and anti-LFA-1 along with anti-CD8 and MHC-I tetramer. Representative plots for CD44, CD62L, CCR7 and LFA-1 are gated on tetramer-binding CD8 T cells. The numbers represent the MFI for the indicated protein. (C) Cytokine production by LCMV-specific memory CD8 T cells from +/+ and FOXO3L mice. Representative dot plots (left) are gated on total lymphocytes. The top numbers indicate the MFI for IFNγ while the bottom number in the plot indicates the percentage of total splenocytes that are CD8 and IFNγ positive. Dot plots (right) represent the percentage of IL-2 and/or TNFα producing cells among IFNγ+ve CD8 T cells. The plots represent data from 1 of 2 independent experiments with 4–7 mice/group/experiment. (D) +/+ and FOXO3L mice were infected with LCMV and given a BrdU injection at 120 days PI and administered BrdU in drinking water between days 120–128 PI (top). At the end of the BrdU pulse, splenocytes were stained with anti-CD8, MHC-I tetramers and anti-BrdU (top) or anti-Ki67 (bottom). The percentage of BrdU or Ki67 positive cells amongst tetramer positive CD8 T cells was determined by flow cytometry. Bars represent data from at least 5 mice/group.