Figure 6.

GDNPs may elicit macrophages to secrete chemokines for T cell chemotaxis

(A) CD8⁺ T trafficking and biodistribution of DiI-labeled CD8⁺ T lymphocytes in CT26 murine colon tumor with BMDMs + PBS or BMDMs + GDNPs treatment administered intravenously (scale bar, 50 μm; n = 5 for each group, ∗∗p < 0.01). (B) Bubble plots showing correlations between Cd8a and chemotactic gene-related transcripts in COAD and TNBC from TCGA cancer datasets. (C) Volcano plot showing upregulated chemotactic gene expression from results of RNA sequencing for M2-BMDMs + GDNPs or M2-BMDMs + PBS (p < 0.05, fold change (FC) > 1.2, three samples each group). (D) Relative gene expressions of Ccl3, Ccl5, Cxcl9, and Cxcl10 in M2 like-BMDMs or GDNPs-stimulated M2-BMDMs for 12 h/24 h by real-time PCR (n = 3−4 for each group, ∗∗∗∗p < 0.0001). (E) CCL5 and CXCL9 concentration in the culture media of M2-BMDMs, GDNPs + M2-BMDMs for 12 h, and GDNPs + M2-BMDMs for 24 h by ELISA (n = 4 for each group, ∗p < 0.05, ∗∗∗p < 0.001). (F) Schematic diagram and quantification of chemotactic assay. CFSE-stained CD8⁺ T cells migrated toward the supernatant from TAM culture media of the Combo group in the presence of anti-CCL5 and anti-CXCL9 neutralization by flow cytometry (n = 3 for each group, ∗∗p < 0.01). For all panels, data are presented as mean ± SEM and analyzed using one-way ANOVA or Student’s t test.