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. 2022 Sep 30;14(10):2157. doi: 10.3390/v14102157

Figure 7.

Figure 7

Generation of CD20-CAR-T cells with Ad-LVs. CAR-T cells were generated from activated Pan T cells using Ad-LV (0.5 TU/cell). To generate CAR-T cells with varying CD4/CD8 ratios, either LLE-CD4 f(ab)2 or LLE-CD8 f(ab)2 adapters alone or a mixture of both adapters in 1:1 or 1:5 ratio was used. (A) Representative data of T cells transduced with Ad-LVs using LLE-CD4 f(ab)2 (clone: MT-466) or LLE-CD8 f(ab)2 (clone: BW135/80) alone or in a mixture are shown. (B) The CD4/CD8 ratio of the CAR-T cells is shown. To analyze if the generated CAR-T cells are functional co-cultures with CD20, + GFP+ Raji cells were set up in an E:T ratio of 1:1. (C) Specific lysis of target cells was analyzed via flow cytometry 24 h post setup of the co-culture. (D) Secretion of specific cytokines was analyzed using the MACS Plex Assay Kit. (E) Activation/exhaustion marker expression was analyzed 6 days post setup of the co-culture assay. Data are represented as mean ± SD of 4 different donors. Statistics: repeated measure one-way ANOVA using Dunnett’s or Tukey’s multiple comparison test; ns = non-significant * p < 0.05, ** p < 0.01.