Journal: Nature Communications

Article Title: Cell microparticles loaded with tumor antigen and resiquimod reprogram tumor-associated macrophages and promote stem-like CD8 + T cells to boost anti-PD-1 therapy

doi: 10.1038/s41467-023-41438-9

Figure Lengend Snippet: a Schematic schedule for the antitumor experiment of combination of R848@M2pep-MPs AFP and anti-PD-1 antibody in orthotopic Hepa1-6 tumor-bearing mice. b , c Tumor images ( b ) and tumor weight ( c ) of orthotopic Hepa1-6 tumor-bearing mice after treatment with R848@M2pep-MPs AFP in the presence or absence of anti-PD-1 antibody at the anti-PD-1 antibody dosage of 5 mg kg −1 and R848 dosage of 0.5 mg kg −1 indicated in ( a ). Data are presented as means ± s.d. for ( c ). (n = 5 mice per group; one-way ANOVA followed by Tukey’s HSD post-hoc test). d Kaplan–Meier survival plot of orthotopic Hepa1-6 tumor-bearing mice after treatment indicated in ( a ). (n = 8 mice per group). e – l The numbers of CD8 + T cells ( e ), CD8 + Ki67 + T cells ( f ), CD8 + CD69 + T cells ( g ), CD8 + IFNγ + T cells ( h ), CD8 + GzmB + T cells ( i ), CD8 + PD-1 + TCF-1 + T cells ( j ), CD8 + PD-1 + TCF-1 - T cells ( k ) and CD8 + PD-1 + TCF-1 - GzmB + T cells ( l ) in tumor tissues of orthotopic Hepa1-6 tumor-bearing mice after treatment indicated in ( a ). Data are presented as means ± s.d. (n = 5 mice per group; one-way ANOVA followed by Tukey’s HSD post-hoc test). m Schematic schedule for ELISPOT and T cell cytotoxic assay. n , o Images ( n ) and numbers ( o ) of IFNγ immune spots from OVA 257-264 - or AFP 212 -restimulated splenocytes isolated from orthotopic Hepa1-6 tumor bearing mice after treatment indicated in ( m ) by the ELISPOT assay. Data are presented as means ± s.d. (n = 5 biological independent samples; two-way ANOVA followed by Bonferroni’s multiple comparisons post-test). p Cytotoxicity of T cells against Hepa1-6 cells when T cells isolated from OVA 257-264 - or AFP 212 -restimulated splenocytes were incubated with Hepa1-6 cells at the effector/target ratio of 10:1 for 6 h as indicated in ( m ) by LDH assay. Data are presented as means ± s.d. (n = 5 biological independent samples; two-way ANOVA followed by Bonferroni’s multiple comparisons post-test). Source data are provided as a Source Data file.

Article Snippet: For ELISPOT evaluation, 5 × 10 5 cells splenocytes isolated from the orthotopic Hepa1-6 tumor bearing mice after treatment were seeded into the wells of ELISPOT plates (Biolegend, San Diego, CA, USA) and incubated with 10 μg mL −1 AFP 212 or OVA 257-264 peptide at 37 °C in 5% CO 2 for 24 h. The cells were lysed and the amounts of IFNγ spot-forming cells were determined by an automated ELISPOT Plate Reader (ASTORTM, Mabtech, Sweden).

Techniques: Enzyme-linked Immunospot, Isolation, Incubation, Lactate Dehydrogenase Assay