Fig. 6

HK2 is largely dispensable for immunity to LCMV. Eight to twelve-week old WT and T-Hk2^−/− mice were injected intraperitoneally with 2 × 10⁵ pfu LCMV. a–c T cell expansion was monitored over time by flow cytometry of peripheral blood. a Number of CD8+ cells. b Percentage of CD8+ cells positive for gp-33 tetramer (D(b)/LCMV.GP33.KAVYNFATM) or (c) CD44. Biological replicates, n = 6 WT, 14 T-Hk2^−/− mice from three independent experiments. d Number of gp33-tetramer+ CD8+ T cells in spleen of WT and T-Hk2^−/− mice at 60 DPI. Biological replicates, n = 6 WT, 12 T-Hk2^−/− mice from two independent experiments, analyzed by unpaired Student’s t test. e Viral load as measured by plaque assay from right lobe of liver from 3 and 8 DPI. Biological replicates, n = 5 mice from two independent experiments. f Splenocytes from uninfected mice and mice 60 DPI were stimulated in vitro for 5 h with 30 ng/mL gp_33–41 peptide or vehicle control and stained for IFNγ and TNFα secretion. Example gating shown on right. Biological replicates, n = 4 mice from two independent experiments. g, h RNA and metabolites were isolated from sorted splenic CD8+ T cells from WT and T-Hk2^−/− mice 8 DPI. g Volcano plot of gene expression differences. Differentially regulated genes include those with greater than 2 fold change in expression and multiplicity adjusted p value < 0.01. h Volcano plot of metabolite concentration differences. Vertical dashed lines indicate 2 fold change in metabolite concentration. Horizontal dashed line indicates no discoveries made (False Discovery Rate of 0.1, two-stage Benjamini-Krieger method, n = 5 mice). In all panels unless otherwise noted, error bars represent mean ± SEM, two-way ANOVA with Sidak multiple comparisons correction (a–c includes repeated measures), *p < 0.05; **p < 0.01. NS = not significant. n.d. = not detected. DPI = days post injection