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. 2021 Jan 8;13(2):197. doi: 10.3390/cancers13020197

Figure 2.

Figure 2

Bcl-2 as a potential enhancer of CAR-T cell survival. (A) Schematic diagram of the CD20- targeting CAR constructs used in the study. An anti-human CD20 single-chain variable fragment (scFv) was linked to 4-1BB and CD3ζ to generate the 20BBZ construct. Various survival-promoting molecules were linked to CD3ζ via a porcine teschovirus-1 2A (P2A) peptide. (B) Flow cytometry analysis of CAR expression on the indicated CAR-T cells. (C) Comparison of long-term proliferation of 20BBZ CAR-T cells and 20BBZ CAR-T cells with the indicated anti-apoptotic molecules. Arrows indicated the irradiated Raji stimulation (effector to target (E:T) = 3:1). (D) Overall expansion of CAR+ T cells in long-term cultures of CD20BBZ CAR-T cells and CD20BBZ-Bcl-2 CAR-T cells. Arrows indicated the irradiated Raji stimulation (E:T = 3:1). (E) 20BBZ and 20BBZ-Bcl-2 CAR-T cells proliferation on 22–28 days after long-term culture. Each dot presented one donor. Representative results from one of four replicate experiments are shown (B–D). Statistical significance was determined by unpaired t-test (D) or paired t-test (E). Statistical significance was presented by *** p < 0.001 or as indicated.