Fig. 5.

T cells contribute to anti-tumor effects achieved with macrophage-programming nanoparticles. a–e Nanoparticle-mediated macrophage programming increases T cell recruitment into tumor lesions. Shown are representative confocal images of peritoneal metastases of ID8 ovarian cancer cells in the mesentery. Tissues were collected after 6 biweekly i.p. injections of PBS or IRF5/IKKβ NPs (50 μg mRNA/dose), and were stained for the indicated lymphocyte- and myeloid-markers (a, c). Tu = Tumor, Mes = Mesentery. Scale bar: 100 μm. b, d Box plots showing fluorescent signals for each phenotypic marker using Halo™ image analysis software. N = 5. The boxes represent the mean values and the line in the box represents median. The bars across the boxes show the minimum and maximum values. Whiskers represent 95% confidence intervals. N = 5 biologically independent samples. e Experimental design for CD8+ T cell depletion studies in C57BL/6 mice with disseminated ID8 ovarian cancer. To deplete CD8+ T cells, mice were injected i.p. with 1 mg anti-CD8b mAb (YTS169.4) three days before the first nanoparticle administration, followed by one 0.5 mg dose every five days for a total of six doses. f Flow cytometry of peritoneal cells collected on day 27 established efficient depletion of CD8+ T cells (average 98.7%). g Plots of ID8 tumor luciferase signal intensities after six nanoparticle injections (day 27). On each box plot, the central mark indicates the median, and the bottom and top edges of the box indicate the interquartile range. Whiskers represent 95% confidence intervals. N = 5 biologically independent samples. Pairwise differences in photon counts between treatment groups were analyzed using the Wilcoxon rank-sum test. Shown are data for five mice per treatment condition pooled from two independent experiments