Figure 5 from NLRP3 Inflammasome Activation Expands the Immunosuppressive Myeloid Stroma and Antagonizes the Therapeutic Benefit of STING Activation in Glioblastoma

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In vivo analysis of intracranial GL261 tumor immune microenvironment phenotypes following STING and NLRP3 inflammasome activation. Mice received intracranial injection of 5.0 × 104 GL261 cells, then were treated with 5 μg of cdGMP and/or nigericin, and tumor-bearing hemispheres harvested 48 hours following treatment for spectral flow cytometry analysis. A, Gr-MDSC, (B) Mono-MDSC, and (C) microglia frequency as a percent of total analyzed CD45+ cells. D, CD206, (E) arginase, and (F) PD-L1 expression on Gr-MDSCs, Mono-MDSCs, and microglia as indicated. Ratios of the number of (G) CD8 T cells/FOXP3+ CD4 Tregs and (H) Gr-MDSC/CD8 T cells. Expression of (I) PD-1 and (J) LAG-3 on CD8 T cells. K, Granzyme expression on CD8 Teff, NK cells, and NKT cells as indicated. Among CD8 Teff, NK cells, and NKT cells, granzyme+ cells were identified and reported as (L) density as cells per tumor-bearing hemisphere and (M) frequency as a percent of total analyzed CD45+ cells. Error bars represent mean ± SEM. Statistical significance was calculated using the Student t test. ns, not significant; *, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001. MFI, mean fluorescence intensity.

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