Fig. 6: Anti-hPD-1 solely blocks, but anti-hPD-L1 and anti-hPD-L2 cooperatively block the binding between hPD-1 and hPD-1 ligands. | Nature Communications

Fig. 6: Anti-hPD-1 solely blocks, but anti-hPD-L1 and anti-hPD-L2 cooperatively block the binding between hPD-1 and hPD-1 ligands.

From: Evaluation of therapeutic PD-1 antibodies by an advanced single-molecule imaging system detecting human PD-1 microclusters

Fig. 6: Anti-hPD-1 solely blocks, but anti-hPD-L1 and anti-hPD-L2 cooperatively block the binding between hPD-1 and hPD-1 ligands.The alternative text for this image may have been generated using AI.

a The cells were imaged as shown in Fig. 1c 2 min after contact to an MCC88-103-prepulsed SLB containing I-Ek–, mICAM-1–, hPD-L1–, and hPD-L2–GPI in the absence (top) or presence of pembrolizumab (anti-hPD-1, row 2), nivolumab (anti-hPD-1, row 3), 29E.2A3 (anti-hPD-L1, row 4), 24 F.10C12 (anti-hPD-L2, row 5), or both 29E.2A3 and 24 F.10C12 (bottom) at a concentration of 10 μg/ml. hPD-1-EGFP, green; TCR, red. b The graph shows the percentages of T cells forming hPD-1 microclusters in a (n = 30). c The cells in a were conjugated with an MCC88-103-prepulsed (1 μM) DC-1 cells expressing both hPD-L1-SNAP-tag (red) and hPD-L2-HaloTag (cyan) plus antibodies as in a and images 2 min after contacts. d The cells in a were cocultured for 16 h with 1 μM MCC88–103 and DC-1 cells not expressing or expressing both hPD-L1 and hPD-L2 plus antibodies as in (a). IL-2 in each supernatant was measured by enzyme-linked immunosorbent assay (ELISA). ns not significant. All data are representative of three independent experiments. Bars, 5 μm. Data are presented as mean values ± SD. Statistical analysis was performed by one-way ANOVA. ***p < 0.001, ****p < 0.0001. Source data for b and d are provided as a Source Data file.

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