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. 2015 Apr 28;4(4):e237. doi: 10.1038/mtna.2015.11

Figure 3.

Figure 3

Anti-PD-1 DNA aptamer MP7 antagonizes PD-1/PD-L1 mediated suppression of IL-2 secretion in vitro. (a) IL-2 ELISPOT assay to compare the effects of PD-L1.Fc or an isotype matched control Fc on the IL-2 secretion by splenocytes stimulated with anti-CD3 antibody. PD-L1.Fc (15 µg/ml) reduced IL-2 SFU by 51% as compared to the Fc control. Each bar represents the mean SFU/2 × 105 cells from at least three replicate wells ± SEM. **P < 0.05 relative to anti-CD3 + Fc-control group. (b) Phosphate-buffered saline, cSeq, anti-PD-1 aptamers (250 nmol/l) or an antagonistic anti-PD-1 antibody (RMPI-14 mAb, 125 nmol/l) were added to splenocytes in wells coated with anti-CD3 + PD-L1.Fc to monitor the blockage of PD-L1-mediated suppression of IL-2 secretion. Bars represent % Blockage of PD-L1 suppressed IL-2 Secretion where the IL-2 spot forming units (SFU) in wells without aptamer/antibody are set to 0% and IL-2 SFU in wells supplemented with anti-PD-1 blocking antibody are set to 100% as anti-PD-1 antibody restored IL-2 secretion to levels 20% above that of anti-CD3 stimulation alone. Each histogram bar represents the mean value ± SEM (n = 5). NS, not significant.