Fig. 6
CD4+ T-cell cytokine production in M. tuberculosis infected lungs after FTY720 treatment. 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. Naive and mice immunized with X31-p25 were treated with FTY720 and challenged with M. tuberculosis as in Fig. 5. Lung cells were stimulated with p25 peptide and the cytokine production assessed by ICS assay. a Frequency of cytokine-producing CD4+ T cells in the lung parenchyma at 17 days post M. tuberculosis infection. Cytokine production by GFP-P25 CD4+ T cells in the lung parenchyma at 17 days (b) and 28 days (c) post M. tuberculosis infection. Pie charts represent the relative proportions of p25-specific CD4+ T cells expressing the possible combinations of IFNγ, TNF, and IL-2. Analysis of the different cytokine-producing subsets was performed by flow cytometry using FlowJo Bolean gating tool. Results are shown as mean + SD (n ≥ 5) and are 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
