Suppressive activity of PMN in TB mice. (A) Antigen-specific suppressive activity of PMN-MDSCs isolated from spleens and tumors of LLC TB mice (n = 8 mice per group). Changes from T cell proliferation in the absence of PMNs (100%) are shown. (B) Representative flow cytometric analysis of proliferation of CD8+ T cells upon stimulation with cognate antigen with or without PMNs isolated from tumors of LLC TB mice. (C) Proliferation of antigen-specific T cells in the presence of different populations of PMNs isolated from tumors of LLC TB mice. Changes from T cell proliferation in the absence of PMNs (100%) are shown. n = 9 mice per group. (D) Suppressive activity of PMNs isolated from spleen and tumors of EL-4 TB mice. PMNs were sorted from spleens and tumors of EL-4 TB mice 3 wk after tumor inoculation. PMNs were added to splenocytes from PMEL mice at a 1:1 ratio in the presence of cognate peptide. Proliferation of T cells was measured as dilution of Cell Trace by flow cytometry (n = 9). (E) Frequency of PMN populations in BM of naive (n = 4) and TB (n = 4) mice. (F) Frequency of PMN populations after stimulation of BM-derived PMNs with TES in normal and hypoxic conditions (n = 6). (G) PMNs were isolated using magnetic beads from BM of naive mice and were cultured with different concentrations of GM-CSF or 20% TES, with or without GM-CSF neutralizing antibody. Frequency of PMNs was analyzed after 24 h by flow cytometry (n = 4). (H) Frequency of populations of PMNs in spleen and tumors of LLC TB WT, S100A9Tg, or S100A9KO mice (n = 3). All data in the figure are presented as mean ± SD. For comparisons between groups, one-way ANOVA with correction for multiple comparisons was used. *, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001.
Figure 6.

Suppressive activity of PMN in TB mice. (A) Antigen-specific suppressive activity of PMN-MDSCs isolated from spleens and tumors of LLC TB mice (n = 8 mice per group). Changes from T cell proliferation in the absence of PMNs (100%) are shown. (B) Representative flow cytometric analysis of proliferation of CD8+ T cells upon stimulation with cognate antigen with or without PMNs isolated from tumors of LLC TB mice. (C) Proliferation of antigen-specific T cells in the presence of different populations of PMNs isolated from tumors of LLC TB mice. Changes from T cell proliferation in the absence of PMNs (100%) are shown. n = 9 mice per group. (D) Suppressive activity of PMNs isolated from spleen and tumors of EL-4 TB mice. PMNs were sorted from spleens and tumors of EL-4 TB mice 3 wk after tumor inoculation. PMNs were added to splenocytes from PMEL mice at a 1:1 ratio in the presence of cognate peptide. Proliferation of T cells was measured as dilution of Cell Trace by flow cytometry (n = 9). (E) Frequency of PMN populations in BM of naive (n = 4) and TB (n = 4) mice. (F) Frequency of PMN populations after stimulation of BM-derived PMNs with TES in normal and hypoxic conditions (n = 6). (G) PMNs were isolated using magnetic beads from BM of naive mice and were cultured with different concentrations of GM-CSF or 20% TES, with or without GM-CSF neutralizing antibody. Frequency of PMNs was analyzed after 24 h by flow cytometry (n = 4). (H) Frequency of populations of PMNs in spleen and tumors of LLC TB WT, S100A9Tg, or S100A9KO mice (n = 3). All data in the figure are presented as mean ± SD. For comparisons between groups, one-way ANOVA with correction for multiple comparisons was used. *, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001.

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