Figure 6
Figure 6. Extended duration of IL-7 treatment during the clonal contraction phase alters the phenotypic attributes of LCMV-specific CD8 T cells. Mice were infected with LCMV-Clone 13 and treated with either IL-7 or PBS between days 8 and 30 PI, as described in Figure 3. At day 90 PI, splenocytes were stained with anti-CD8, anti-CD44, anti-CD122, anti-CD127, anti-CD62L, anti-KLRG-1, anti–Cbl-b, anti-PD-1, and Db/GP33 tetramers (A-C). (D) At day 44 PI (14 days after cessation of IL-7 therapy), splenocytes were incubated with anti-CD107a (anti–LAMP-1) and DbGP33 tetramer and stimulated with LCMV-specific cognate peptide for 1 hour at 37°C. After incubation, cells were washed and stained with anti-CD8 antibody. Data were analyzed by flow cytometry, and FACS plots were gated on tetramer-binding CD8 T cells. The numbers in panels A and B are the mean fluorescence intensity (MFI) and/or percentages of tetramer-binding CD8 T cells. Data are from an analysis of 4-5 IL-7–treated (black line) or PBS-treated (gray line) mice. Stainings with anti–Cbl-b antibodies preincubated with the specific immunogenic Cbl-b peptide are shown as dotted lines. *P ≤ .05; **P ≤ .005; ***P ≤ .001.

Extended duration of IL-7 treatment during the clonal contraction phase alters the phenotypic attributes of LCMV-specific CD8 T cells. Mice were infected with LCMV-Clone 13 and treated with either IL-7 or PBS between days 8 and 30 PI, as described in Figure 3. At day 90 PI, splenocytes were stained with anti-CD8, anti-CD44, anti-CD122, anti-CD127, anti-CD62L, anti-KLRG-1, anti–Cbl-b, anti-PD-1, and Db/GP33 tetramers (A-C). (D) At day 44 PI (14 days after cessation of IL-7 therapy), splenocytes were incubated with anti-CD107a (anti–LAMP-1) and DbGP33 tetramer and stimulated with LCMV-specific cognate peptide for 1 hour at 37°C. After incubation, cells were washed and stained with anti-CD8 antibody. Data were analyzed by flow cytometry, and FACS plots were gated on tetramer-binding CD8 T cells. The numbers in panels A and B are the mean fluorescence intensity (MFI) and/or percentages of tetramer-binding CD8 T cells. Data are from an analysis of 4-5 IL-7–treated (black line) or PBS-treated (gray line) mice. Stainings with anti–Cbl-b antibodies preincubated with the specific immunogenic Cbl-b peptide are shown as dotted lines. *P ≤ .05; **P ≤ .005; ***P ≤ .001.

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