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Fig. 1 | Journal for ImmunoTherapy of Cancer

Fig. 1

From: Intratumoral immunotherapy with TLR7/8 agonist MEDI9197 modulates the tumor microenvironment leading to enhanced activity when combined with other immunotherapies

Fig. 1

MEDI9197 activates innate and adaptive immune cells. a SEAP reporter activity in HEK293-NFκB-SEAP cells expressing human TLR7 or TLR8. Results are shown as fold-change relative to DMSO. b pDC and mDC enriched from peripheral blood and treated with MEDI9197 or DMSO. IFN-α and IL-12p40 secretion was measured from pDC and mDC, respectively, by ELISA. Results are shown as mean ± SEM, n = 3 donors. c IL-12p70 cytokine release from 20 ng/mL LPS or 3 μM MEDI9197-treated monocyte derived macrophages was tested by ELISA. Results show the mean of 5 donors. d Median fluorescence intensity (MFI) of activation markers on cell subsets from human PBMCs stimulated with MEDI9197 or DMSO. NK cells = CD3 CD19 CD56high; B cells = CD3 CD19+; pDC = CD3 CD19 CD20 CD56 HLA-DR+ CD14 CD16 CD123+ BDCA4+; monocytes = CD3 CD19 CD20 CD56 HLADR+; Data show the mean ± SEM, representative of 4 donors. e IL-5 release (ELISA) from human PBMCs stimulated with MEDI9197 or DMSO with or without PHA-L. Data show mean of technical triplicates ±SEM, representative of 9 donors. f Cytokine production (ELISA and MSD) from human PBMCs stimulated with MEDI9197, C class CpG, or STING agonist. Results show mean ± SEM of 3 donors, representative of 6 donors. g Percent specific killing of Eu-loaded K562 target cells co-cultured with NK cells isolated from human PBMCs stimulated with MEDI9197 or DMSO. Data show mean of technical duplicates ±SEM, 9 donors. h and i Proportion of CMV peptide-specific CD8 cells (h) or granzyme B-positive CD8 cells (i) after stimulation of PBMCs with MEDI9197 or DMSO and a titration of CMV peptide, 2 donors. Data are representative of ≥2 independent experiments. Statistical analyses were performed by one-way (C) and two-way (F-I) ANOVA with Tukey’s (C, F, H, I) or Sidak’s (G) post hoc test or Welch’s upaired T test (one-tailed) (B, D). *p < 0.05, **p < 0.01, ***P < 0.001, ****p < 0.0001

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