Fig. 2: IBI319 blocks the PD-1/PD-L1 interaction and selectively enhances CD137 activity in a PD-1-dependent manner. | Nature Communications

Fig. 2: IBI319 blocks the PD-1/PD-L1 interaction and selectively enhances CD137 activity in a PD-1-dependent manner.

From: Cancer immune therapy with PD-1-dependent CD137 co-stimulation provides localized tumour killing without systemic toxicity

Fig. 2

a PD-1/PD-L1 blocking activities of different antibodies presented as NFAT-mediated luciferase activity. Fold induction was calculated as the mean relative light units (RLU) of tested antibodies/mean RLU of the no antibody control. b CD137 agonist activities of different antibodies presented as NFκB-mediated luciferase activity. Jurkat-CD137-NFκB-Luc (Jurkat-CD137) cells were co-cultured with two times the number of CHO-S-PD-1 cells (left), without CHO-S-PD-1 cells (middle), and with different amounts of CHO-S-PD-1 cells (right). Fold induction was calculated as the mean RLU of tested antibodies/mean RLU of the no antibody control. c Three dimensional (3D) and 2 dimensional (2D) cell culture systems to test the cis- and trans-cell crosslinking of IBI319. (Left) The 3D and 2D cell culture models when treated with IBI319. (Right) The relative luciferase signals of the Jurkat-CD137-NFκB-Luc2P-PD-1 alone and Jurkat-CD137 + CHO-S-PD-1 groups were calculated with the equation: (mean RLU of tested antibodies in 3D culture/mean RLU of the control in 3D culture)/(mean RLU of tested antibodies in 2D culture/mean RLU of the no antibody control in 2D culture). d Relative IFN-γ production in an MLR assay. Antibodies were 4-fold serially diluted from 200 nM. e Relative IFN-γ production level in a T cell co-stimulation assay. Primary T cells were treated with 1 μg/mL CD3/CD28 activator, and antibodies were serially diluted 10-fold from 400 nM in the presence of CHO-S or CHO-S-PD-1 cells. f IFN-γ production level in a cytomegalovirus (CMV) T cell memory recall assay. PBMCs from a donor previously infected with CMV were co-cultured with a CMV peptide pool and antibodies at 50 nM for 4 days (data for three donors in Supplementary Fig. 2e). a and b show two technical replicates of one representative experiment from three independent experiments. mean and SD. c, d, e, f show individual values of three independent experiments and the mean ± SD. Statistics were performed in the indicated groups: two-way ANOVA with Tukey’s multiple comparisons test (b) and two-sided paired t-test (c, d, e, f); p values are indicated in the graphs.

Back to article page