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. 2023 May 19;11(5):e006714. doi: 10.1136/jitc-2023-006714

Figure 2.

Figure 2

IM-T9P1-ASO stimulates in vitro DC activation and IL-12 secretion while decreasing PD-L1 expression. (A–B) Flow cytometry analysis of PD-L1 expression in in vitro-cultured moDCs exposed to Ctr-ASO or IM-T9P1-ASO (n=4 cell cultures/condition). Representative histogram plots (A) and cumulative analysis of PD-L1 MFI (B). (C–D) Flow cytometry analysis of activation markers in in vitro-cultured FL-cDCs derived from wild-type mice (C and D), or transgenic TLR9–/– and PD-L1–/– mice (D), and exposed to Ctr-ASO or IM-T9P1-ASO (n=3–4 cell cultures/condition). (E–F) Flow cytometry analysis of CD8+ OT-I cell proliferation after 3 days of culture with OVA-loaded moDCs previously exposed to the indicated agents. (E) Representative histograms showing CellTrace Violet (CTV) dilution. (F) Quantification of the data (n=4 cell cultures/condition). (G) ELISA-based quantification of granzyme B in medium conditioned by CD8+ OT-I cells treated as in (E–F). Data are presented as mean±SEM. For comparisons between two groups, Student’s two-tailed t-test was used. For comparisons between multiple groups or variables, one-way (C, F, G) or two-way (D) analysis of variance was used. *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001. ASO, antisense oligonucleotides; cDC, conventional DC; CpG, cytosine-phosphate-guanine; DC, dendritic cell; IL, interleukin; mAb, monoclonal antibody; MFI, mean fluorescence intensity; MHC, major histocompatibility complex; moDCs, monocyte-derived dendritic cells; PD-L1, programmed cell death ligand 1; WT, wild-type; TLR9, toll-like receptor 9.