Fig. 5

α-CD3/α-CD28 npGG can stimulate T cells activation A Activation profile of murine splenocytes when stimulated with varying concentrations of npGG aAPCs. I. Measurement of CD4⁺ T-cell proliferation in vivo by CFSE dilution. Murine splenocytes were labelled CFSE and then stimulated with both α-CD3 and α-CD28 npGG in a ratio of 1:1 for a period of 7 days. After this period cells were labelled and gated for anti-CD4 and analyzed on a BD FACSAria™ III. Results are presented as the percentage of cells in the final population that have divided. Data are presented as means ± standard error. *, #,γ p < 0.05; **, ##, γγ p < 0.01; ***, ###,γγγ p < 0.001 relatively to unmodified particles at 10 μg, 50 μg or 100 μg respectively. II. IL-2 production by murine splenocytes when stimulated over 7 days with both anti-CD3 and anti-CD28 GG particles in a ratio of 1:1. Control (CTRL) corresponds to the conditions in which unmodified particles were used to stimulate the cell cultures. The IL-2 contents of the control media was undetectable. Data are presented as means ± standard error. *, ϕ, Δ, γ p < 0.05; **, ϕ ϕ, Δ Δ, γγ p < 0.01; ***, ϕ ϕ ϕ, Δ Δ Δ, γγγ p < 0.001; ****, ϕ ϕ ϕ ϕ, Δ Δ Δ Δ, γγγγ p < 0.0001. * was used when pairwise comparisons are performed relative to unmodified particles at 10 μg, 50 μg or 100 μg respectively. ϕ was used when comparisons are performed relative to activator beads, Δ relative to unstimulated cells and γ relatively to EDC at comparative 10 μg or 20 μg of antibody at 10 μg, 50 μg or 100 μg respectively. B qPCR mRNA fold change of (i) immune checkpoint genes PDCD1 and CTLA4 (ii) cytotoxic genes PRF1 and GZMB in murine splenocytes stimulated with activator-npGG over 7 days. Quantitative results are expressed as the mean ± standard deviation where n = 3, * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001. One-way ANOVA with Kruskal–Wallis multiple comparison post-test were used