Fig. 4: Reduced bacterial loads and lung inflammation without restoration of Mtb-specific T cell responses in gemcitabine (Gem)-treated mice upon depletion of Ly6G+ cells. | Nature Communications

Fig. 4: Reduced bacterial loads and lung inflammation without restoration of Mtb-specific T cell responses in gemcitabine (Gem)-treated mice upon depletion of Ly6G+ cells.

From: Mycobacterium tuberculosis-specific T cells restrain anti-cancer drug-induced neutrophilic lung inflammation in tuberculosis

Fig. 4: Reduced bacterial loads and lung inflammation without restoration of Mtb-specific T cell responses in gemcitabine (Gem)-treated mice upon depletion of Ly6G+ cells.The alternative text for this image may have been generated using AI.

A 28 days post Mtb infection, equal numbers of isolated CD11b+Ly6G+ cells from Mtb-infected mice (~350 CFU) and Mtb-infected mice treated with Gem were lysed with 0.05% Triton X-100, and the lysates were plated onto 7H10 agar to enumerate the bacteria (n = 4). The graph shows the mean ± SD. Data were analyzed by two-tailed Mann–Whitney rank test. B At 14, 16, and 18 days post Mtb infection, a monoclonal anti-Ly6G antibody (α1A8) or isotype control was administered i.p. into Gem treated mice. Then, mice were sacrificed at 21 and 28 days post Mtb infection. C, D Immunofluorescence and H&E staining of the lungs at 28 days post Mtb infection (10×, scale bars = 2.0 mm; 400×, scale bars, 50 μm). E The percentages of inflamed area in the lungs at 28 days post Mtb infection (n = 4). The graph shows the mean ± SD. Data were analyzed by two-tailed Mann–Whitney rank test. F CFUs in the lungs at 28 days post Mtb infection, determined by counting the viable bacteria (n = 4). The graph shows the mean ± SD. Data were analyzed by two-tailed Mann–Whitney rank test. G The levels of cytokines in lung homogenates obtained from 28 days post Mtb infection were analyzed by ELISA (n = 4). The graph shows the mean ± SD. Two-tailed Mann–Whitney rank tests were used to compare groups. n.s. not significant, *p < 0.05. H The frequencies of IFN-γ- and TNF-producing CD4+ T cells were analyzed by flow cytometry. The graph shows the mean ± SD from pooled samples (n = 3) from each group (n = 4). Data were analyzed by one-way ANOVA with post hoc Tukey test. n.s. not significant, **p < 0.01, ***p < 0.001, and ****p < 0.0001 between group. I The level of G-CSF in serum at 28 days post Mtb infection were analyzed by ELISA (n = 4). The graph shows the mean ± SD. Two-tailed Mann–Whitney rank tests were used to compare groups. n.s. not significant, *p < 0.05. A, G–I The data are representative of a single experiment or C–F at least two independent experiments. Source data are provided as a Source Data file.

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