Skip to main content
. 2022 Apr 11;21(6):948–959. doi: 10.1158/1535-7163.MCT-21-0802

Figure 4.

Figure 4. Adenosine signaling results in a dominant suppression of T-cell activation in the presence of PD-1 blockade that can be restored by blocking CD73 with AB680. A–D, Human moDCs were cultured with CD4+ T cells in an allogeneic MLR for 96 hours in the presence of α-PD-1, AMP and AB680 as indicated. A and B, Gene expression measured by quantitative RT-PCR from an MLR treated with IgG4 isotype (0.67 nmol/L), anti-PD-1 (0.67 nmol/L), AMP (100 μmol/L), and AB680 (100 nmol/L) as indicated for 72 hours. Data shown from one representative donor pair. Similar results obtained in another donor pair. C, Secretion of IFNγ was measured to determine T-cell functionality after 96 hours treatment with IgG4 isotype, anti-PD-1 (0.67 or 6.7 nmol/L), AMP (100 μmol/L) ± AB680 (100 nmol/L) as indicated. Data shown of IFNγ in one donor pair (left) and of normalized IFNγ of 16 donor pairs from three independent experiments. Each symbol represents a unique donor pairing. D, Secretion of IFNγ was measured after 96 hours treatment with IgG4 isotype or anti-PD-1 (0.67 or 6.7 nmol/L) ± the addition of AMP (100 μmol/L) and AB680 (100 nmol/L) at the beginning of the coculture or after 48 hours. Data shown of IFNγ in one donor pair (left) and normalized IFNγ of nine donor pairs from two independent experiments. Data represented as mean ± SEM. *, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001, (A–C) Dunnett multiple comparisons test versus AMP + α-PD-1. D, Sidak multiple comparisons test.

Adenosine signaling results in a dominant suppression of T-cell activation in the presence of PD-1 blockade that can be restored by blocking CD73 with AB680. A–D, Human moDCs were cultured with CD4+ T cells in an allogeneic MLR for 96 hours in the presence of anti–PD-1, AMP and AB680 as indicated. A and B, Gene expression measured by quantitative RT-PCR from an MLR treated with IgG4 isotype (0.67 nmol/L), anti–PD-1 (0.67 nmol/L), AMP (100 μmol/L), and AB680 (100 nmol/L) as indicated for 72 hours. Data shown from one representative donor pair. Similar results obtained in another donor pair. C, Secretion of IFNγ was measured to determine T-cell functionality after 96 hours treatment with IgG4 isotype, anti–PD-1 (0.67 or 6.7 nmol/L), AMP (100 μmol/L) ± AB680 (100 nmol/L) as indicated. Data shown of IFNγ in one donor pair (left) and of normalized IFNγ of 16 donor pairs from three independent experiments (right). Each symbol represents a unique donor pairing. D, Secretion of IFNγ was measured after 96 hours treatment with IgG4 isotype or anti–PD-1 (0.67 or 6.7 nmol/L) ± the addition of AMP (100 μmol/L) and AB680 (100 nmol/L) at the beginning of the coculture or after 48 hours. Data shown of IFNγ in one donor pair (left) and normalized IFNγ of nine donor pairs from two independent experiments (right). Data represented as mean ± SEM. *, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001, (A–C) Dunnett multiple comparisons test versus AMP + anti–PD-1. D, Sidak multiple comparisons test.