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

Figure 6.

Figure 6

Anti-PD-1 aptamer PEG-MP7 is not cytotoxic and enhances tumor-specific T-cell responses in vivo without induction of a TLR9-mediated innate immune response. (a) Cell viability assay was quantified using (3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium) (MTS) dye after a 24-hour incubation of MC38.CEA cells with 5 µmol/l of PEG-MP7, PEG-cSeq, MP7, or 8M urea. Bars represent the mean OD490 nm ± SD from replicate wells. ***P < 0.001. Aptamer-induced TLR-9 signaling was assayed by injection of PEG-cSeq, PEG-MP7, PBS, or a control CpG ODN into naive C57Bl/6 mice (n = 3) and 3 hours later the sera levels of TNFα (b) and IL-6 (c) were quantified by ELISA. Only injection of the CpG ODN yielded signal above the lower limit of detection (dotted line). Each symbol represents the average cytokine levels from each individual animal measured in duplicate.