Figure 1

Significantly fewer CD4⁺ and CD8⁺ lymphocytes accumulate in the infected spleens of p47^phox−/− mice during primary L. monocytogenes infection. WT () and p47^phox−/− () mice were infected with 5 × 10⁴ CFU (0.1 LD₅₀) rLM-OVA. On the indicated post infection day the mice were euthanized and single-cell splenocyte cultures were incubated for 3–5 h with 1 μM OVA_323-339 and/or OVA_257-264 peptides for CD4 and CD8 T lymphocyte stimulation, respectively. Monensin was added for the final 2 h of culture, and the cells were harvested and stained for surface CD4, CD8, and intracellular IFNγ expression. (a) Percentage of CD4⁺ and CD8⁺ lymphocytes, and the percentage of CD4⁺ and CD8⁺ lymphocytes expressing IFNγ. The data are the mean (±S.E.M.) for six individual mice from two separate experiments with three of each genotype/experiment. (b) p47^phox−/− CD8⁺ T lymphocytes effectively kill target cells. On post infection days 3 and 7, as indicated, spleen-derived CD8⁺ T cells were co-cultured with CFSE-labeled EL4 or E.G7-OVA target cells at the indicated effect: target cell ratios for 4 h. The results are presented as percentage of PI⁺ E.G7-OVA or EL4 target cells. Results are the mean (±S.E.M.) of three independent experiments including three of each genotype/experiment. (c) On post infection day 3, the percentage of non-viable EMA⁺ CD4⁺ or EMA⁺CD8⁺ lymphocytes was determined. The data are the mean (±S.E.M.) percentage for five individual mice. *P<0.005, **P<0.05