Fig. 4: PGN4.9 nanoadjuvant reprogrammes M2-like macrophages into M1 phenotype for enhanced antigen cross-presentation in vitro. | Nature Communications

Fig. 4: PGN4.9 nanoadjuvant reprogrammes M2-like macrophages into M1 phenotype for enhanced antigen cross-presentation in vitro.

From: pH-gated nanoparticles selectively regulate lysosomal function of tumour-associated macrophages for cancer immunotherapy

Fig. 4

a AND-gated design of PGN4.9 nanoadjuvant with pH- and enzyme-dual selectivity. b TLR agonistic activity of various IMDQ treatments via a TLR reporter cell assay (n = 6 experiments). c Heatmap of several protein levels from proteomics data of in M1-, M2- and M2-like BMDMs treated with PGN4.9 nanoadjuvant (n = 3 experiments). Scale, (M1 − M2avg)/(M1 + M2avg), (M2 − M1avg)/(M2 + M1avg) or (PGN4.9 − M2avg)/(PGN4.9 + M2avg). d Ratio of CD206 to CD86 expression in BMDMs lysate by flow cytometry (n = 3 experiments). e The expression of iNOS and Arginase in M1-like BMDMs or M2-like phenotype treated with IMDQ preparations for 24 h (n = 3 experiments). f Ratiometric images of M2-like macrophages treated with different IMDQ formulations, followed by the treatment of PGN4.9 nanoreporter (n = 3 experiments). Scale bar, 20 μm. g Schematic illustration of PGN4.9 nanoadjuvant resetting M2-like BMDMs to M1 phenotype for antigen cross-presentation. h The lysosomal pH of M2-like macrophages after incubation with various IMDQ formulations (n = 18 cells for M2 group; n = 19 cells for M1, IMDQ and PGN4.9 groups; n = 20 cells for other groups). i Reduced expression of cathepsin B in M2-like macrophage after treatment with PGN4.9 nanoadjuvant (n = 3 experiments). Scale bar, 20 μm. j Immunoblots of representative cathepsin family (e.g. cathepsin B, L and S) in M2-like BMDMs before and after treatment with PGN4.9 nanoadjuvant (n = 3 experiments). k Representative images of DQ-ovalbumin (green) degradation assays in M2-like BMDMs pretreated with PGN4.9 nanoadjuvant. Scale bar, 20 μm. l CD80+CD86+- and m antigen-positive BMDMs after treatment with different IMDQ formulations and OVA257–280 long peptides for 24 h (n = 3 experiments). Statistical significance was analysed by one-way ANOVA followed by Tukey’s multiple comparisons test. All measurements are presented as mean ± s.d. Source data are provided as a Source Data file.

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