Fig. 5
Effect of FTY720 treatment on CD4+ T cells after infection with M. tuberculosis. Fifty-thousand GFP-P25 Tg splenic cells were adoptively transferred into naive recipient C57Bl/6 mice 1 day prior to intranasal immunization with 5 × 104 pfu X31-p25. Six weeks after immunization, both naive and immunized mice were treated with FTY720, 1 mg/kg by i.p. daily for 20 days and challenged with M. tuberculosis at day 3 of treatment. CD4+ T-cell populations were analyzed and compared with those of naive and immunized mice infected with M. tuberculosis but not treated with FTY720. a Diagram of the experimental plan. The number of CD4+ T cells present in 200 µl of blood (b), mediastinal lymph node (c), lung parenchyma (d), and lung vasculature (e) at 17 days post infection. f Number of GFP-P25 Tg CD4+ T cells, g pie charts representing the proportion of the different subset populations of GFP-P25 CD4+ T cells based on the surface markers CD69, CD11a, CD62L and KLRG1 and h number of CD69+CD11a+CD62LloKLRG1- GFP-P25 CD4+ T cells present in the lung parenchyma at day 17 and 28 post M. tuberculosis infection. Results in b–f and h are shown as mean ± SD (n = 5–7). Results in g were obtained using the FlowJo bolean gating tool. The representative figure shows the results from one of two independent experiments. ****p < 0.0001, ***p < 0.001, **p < 0.01, *p < 0.05, ANOVA followed by Tukey post-test correction
