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Fig. 5 | BMC Biology

Fig. 5

From: Single-cell transcriptomics reveals the effect of PD-L1/TGF-β blockade on the tumor microenvironment

Fig. 5

Combination therapy of anti-PD-L1 plus intratumorally administered CCL5 leads to tumor growth inhibition. a–d Mice bearing s.c. MC38 tumors were administered PBS or CCL5 (1 μg/dose) I.T. three times per a week for 5 doses, and on day 12 tumors were harvested and analyzed by flow cytometry. a tSNE plots of flow data showing the expression of CCR5 within PBS control (left) or CCL5-treated tumors (middle), or where populations are colored according to expression of marker genes as indicated in the key (right). Heat maps showing expression of each flow marker are in Additional file 1, Figure S8. b Left: percent of CCR5+ cells within the CD8+ T cell population (live cells, CD4−, CD8+). Right: percent of CCR5+ cells within the NK cell population (live cells, NK1.1+, CD3−). Black cross bars represent median values. P values were determined using Wilcoxon rank sum test. c Density plots (with outlier cells indicated as dots) of populations from the tSNE analysis in a within control or CCL5-treated samples. Red arrows indicate CD11b+ NK cells (population 23 from a). d Frequency of NK cells (live, single cells, NK1.1+ CD3−) that express CD11b as determined by flow cytometry. Black cross bars represent median values. P value was determined using Wilcoxon rank sum test. e–g Mice bearing s.c. MC38 tumors (n = 12 per group) were administered PBS (I.T., 3 times a week for 3 weeks), CCL5 (I.T., 1 μg/animal, 3 times a week for 3 weeks), aPD-L1 (atezolizumab; I.P., 1 mg/kg, twice a week for 3 weeks), or a combination of CCL5 and aPD-L1. e Average tumor volume ± SEM for each treatment group is shown. P values were determined using Wilcoxon rank sum test, comparing tumor sizes on day 16. f Spider plots showing tumor volume for individual mice over time. g Survival plot for the same study. P values were determined using log-rank test. Non-significant p values are not shown for all panels

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