Fig. 2

Cellular changes in the MB49^luc bladder TME of control Ig– and NHS-muIL12– treated mice at 72 and 144 h after the final NHS-muIL12 treatment. a Representative FACS plots of cytotoxic T lymphocytes (CD8, y-axis) vs myeloid (CD11b, x-axis) cells from control Ig– (left column) and NHS-muIL12–treated (right column) mice bearing MB49^luc bladder tumors. Plots are representative of live CD45⁺ tumor-derived cells at 72 (top row) and 144 h (bottom row) after the final NHS-muIL12 treatment. Numbers within each quadrant represent percent of total cells. In panels b-f, mice bearing MB49^luc bladder tumors and treated with control Ig– (black bars) or NHS-muIL12 (grey bars) were euthanized at 72 and 144 h after the final NHS-muIL12 treatment and the lymphoid/myeloid cellular components within the TME were examined by flow cytometry. Absolute number of each cell type/mg bladder weight for (b) MDSCs: CD11b⁺Gr1⁺F4/80⁻ (n = 9), (c) macrophages: CD11b⁺Gr1⁻F4/80⁺ (n = 9), (d) CD4⁺ T cells (n = 9), (e) CD8⁺ T cells (n = 9), and (f) regulatory T cells (Tregs): CD3⁺CD4⁺FoxP3⁺ (n = 3) are shown. Panels g-j represent ratios of (g) CD8⁺/MDSCs, (h) CD8⁺/macrophages, (i) CD4⁺/MDSCs, and (j) CD4⁺/macrophages at 72 and 144 h after the final NHS-muIL12 treatment (n = 9). Circles and squares represent control Ig–treated and NHS-muIL12–treated mice, respectively. Error bars (panels b-j) represent mean + SEM, Student’s t-test; *P < 0.05. Data are from a representative experiment that was repeated twice with similar results