Figure 5. LCMV-specific T Cells in Cmah−/− Mice Produce a Higher Proportion of Cytokines Than WT T Cells.

Mice infected with acute LCMV Armstrong were sacrificed at day 42, and isolated splenocytes were stimulated for 5 hours with LCMV peptides and stained for multiple cytokines to assay functionality. (A, C–E) Functionality of CD8+ T cells upon LCMV peptide GP276 stimulation. (A) Frequency of Mip1α+IFNγ+ CD8+ T cells upon LCMV peptide stimulation in WT (black triangles in right plot) and Cmah−/− (red squares in right plot) mice. Left and middle plots show the median representative dot plots from each group, while the right plot shows the quantification of the frequencies. (B) Frequency of Mip1α+IFNγ+ CD4+ T cells upon LCMV peptide GP66-77 stimulation in WT (black triangles in right plot) and Cmah−/− (red squares in right plot) mice. Left and middle plots show the median representative dot plots from each group, while the right plot shows the quantification of the frequencies. (C) Frequency of Mip1α+IFNγ+ CD8+ T cells from (A) that are TNFα+IL-2+. Representative plots on the left and middle, quantification on the right as described in (A). (D) Polyfunctional pie charts of day 42 stimulated CD8+ T cells from WT (left pie chart) and Cmah−/− (right pie chart) mice. Red=5 functions, blue=4 functions, orange=3 functions, yellow=2 functions, green=one function. Functions= CD107a, Mip1α, IFNγ, TNFα, and IL-2. (E) Bar graph representation of each combination of the functions defined in (D) between WT (blue bars) and Cmah−/− (red bars) CD8+ T cells, with the corresponding pie chart color on the bottom. *p<0.05, student’s t test. N=5 mice per group. Results are representative of at least two experiments.