Fig. 2: Emapalumab does not affect CD28-based CAR.CD19-T cell in vitro cytotoxicity. | Nature Communications

Fig. 2: Emapalumab does not affect CD28-based CAR.CD19-T cell in vitro cytotoxicity.

From: Neutralizing IFNγ improves safety without compromising efficacy of CAR-T cell therapy in B-cell malignancies

Fig. 2

a Effector T cells (NT or CAR.CD19.CD28) and GFP+ Daudi cells were plated at decreasing E:T ratios, from 1:1 to 1:2.5 either in the presence or absence of 100 μg/ml emapalumab. At the end of the experiments, tumor cells and T cells were collected and assessed by FACS analysis evaluating the residual GFP+ tumor cells. Data are expressed in percentage, as mean ± SD of 3 independent experiments, with effector cells generated from 3 HDs. Data were compared by a two-tailed Student t-test and p value <0.05 was considered statistically significant. b Heatmaps displaying the IFNγ, Granzyme B, IL-2 and TNF-α levels that are secreted by effector T cells (NT or CD28-based CAR.CD19) after 24 h of co-culture with Daudi cells in media containing emapalumab within the concentration range of 0.2–100 μg/ml. Color scale represents cytokine levels as mean of 3 independent experiments with effector cells generated from 3 HDs expressed in percentage. c Real-time kinetics of Daudi elimination exerted by T cells (NT or CAR.CD19.CD28) (1:1 E:T ratio) either in the presence or absence of 100 μg/ml emapalumab. The indicated statistical significance refers to the end point of the experimental assay. Data are expressed as mean ± SD of 3 replicates. Data were compared by a two-tailed Student t-test and p value <0.05 was considered statistically significant. d Proliferation of CAR.CD19.CD28-T cells activated by Daudi cells and monitored for 3 days in the absence (blue area) or in the presence of 100 μg/ml emapalumab (purple area) compared to CAR-T cells at the time of cell plating (day 0, pink area). Exemplificative histograms, relative to one representative experiment, out of the three performed with three different CAR.CD19-T cell donors (p value of the proportion of proliferating CAR.CD19-T cells with vs. without Emapalumab: 0.957). Student’s t-test was employed to calculate statistically significant differences between groups. Source data are provided as a Source Data file.

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