Fig. 1: PVSRIPO treatment of ex vivo tumor tissue induces type-I/III IFN in TAMs.

a Fresh glioblastoma (GBM) tissue was analyzed for distribution of surface CD155 by cell type and responsiveness to innate stimuli. b GBM single-cell suspensions were analyzed for CD155 expression via flow cytometry (n = 13 tumors, see Supplementary Fig. 1 for gating); asterisks denote Bonferroni corrected one-sample t test (p < 0.0064, two-tailed, from left to right: p = 0.0001, 0.0001, 0.0001, 0.007, 0.0005, 0.0008, 0.03, 0.005). Normalized MFI (median florescence intensity) was calculated by subtracting isotype control from stained MFI values for each cell type; mean ± SEM is shown. c, d Tumor slices were treated with PVSRIPO, Poly(I:C), lipopolysaccharide (LPS), or 2′3′-cGAMP as shown; pfu = plaque forming units. Post-treatment cell viability was measured after PVSRIPO treatment [7-AAD staining; n = 6 GBM, n = 3 glioma cell lines (DU54, DU43, and U87; mean of 3 experiments per cell line is shown)] (c) and fold-mock control cytokine release in supernatant was examined after all treatments (d; n = 20 for mock, PVSRIPO, and Poly(I:C); n = 15 for LPS; n = 18 for cGAMP). Only induced cytokines are shown; see Supplementary Fig. 2 for patient-specific induction; Tukey’s post-hoc p < 0.05 (two-tailed) vs mock (*), unless otherwise indicated by bracket, or vs all other groups (#). e Ex vivo slice assay using breast cancer tissue (n = 10) and melanoma tissue (n = 4) testing only PVSRIPO as in (d); (*) one-sample t test p < 0.05 (two-tailed; d: CXCL10 p = 0.003, IFN-λ1 p = 0.02, IFN-α p = 0.03; e: CXCL10 p = 0.03). d, e Violin plots present quartiles and median. f, g Dissociated, post-treatment tumor slices were analyzed by flow cytometry for PD-L1 expression [f, n = 6 GBM, normalized MFI—mock MFI is shown, (*) one-sample t test p < 0.05 (two-tailed): CD14+ CD11b+ PVSRIPO p = 0.02, cGAMP p = 0.02] and intracellular IFIT1 expression for indicated cell types [g, n = 4 GBM and 2 breast cancer, fold-mock control MFI for each cell type is shown, (#) Tukey’s post-hoc test vs all other groups: macrophage vs: endothelial p = 0.0005, tumor p = 0.001, B/T cells p = 0.0004]; see Supplementary Fig. 3 for gating and extended data. h Fresh (n = 3) and previously frozen (n = 6) GBM cell suspensions were subjected to mock or CD14+ depletion. Cell count-normalized mock- and CD14+ -depleted suspensions were treated with PVSRIPO (48 h) and supernatant cytokines were measured; box shows median with quartiles and whiskers indicate range; p-value is from paired t-test (two-tailed) comparing the sum of log(fold-mock control) cytokine values; see Supplementary Fig. S4 for extended data.