FIGURE 6.

P. yoelii induced exacerbation of Tb disease is transient. C57BL/6 mice were infected via the aerosol route with M. tuberculosis H37Rv, and 30 days later with 1 × 10⁵ iRBCs i.p. Lungs and spleens were collected 170 days after P. yoelii infection. (A) Lung and spleen CFU was determined. Data are shown as box and whisker plots with the median and analyzed statistically using Mann–Whitney test (n = 8). (B,C) Superior lung lobes were paraffin-embedded, and 4 μm sections were H&E-stained (one representative image per group is shown). Pulmonary inflammation was quantified using cellˆB software (n = 8; box and whisker plots with the median; Mann–Whitney test). Lung single cell suspensions were analyzed for the presence (D) and function (E) of CD4⁺ and CD8⁺ T cells by flow cytometry (restimulated ex vivo with αCD3/αCD28). (F) Cytokine protein levels were measured in lung lysates in M. tuberculosis and co-infected mice using LEGENDplex. (G–J) Lung cells were gated on CD45⁺ and further on CD19^– and Ly6G^– cells and analyzed for the presence of CD11b⁺ and CD11c⁺ cells. CD11b^–CD11c⁺ cells were further divided into CD11c^int and CD11c^high (H). CD80 and CD86 expression was analyzed in both CD11b^–CD11c⁺ populations (I,J). For full gating strategy, see Supplementary Figures S1A,B,D. Data are shown as means ± SD and were analyzed using the Mann–Whitney test (n = 8).