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. 2022 Jan 27;13:770132. doi: 10.3389/fimmu.2022.770132

Figure 1.

Figure 1

CD45RA+ chimeric antigen receptor (CAR)-T cells showed better transient gene transfer efficiency and expansion capacity, dominant CD8 expression, enriched stem cell memory fraction, and less expression of exhaustion-related markers than CD45RA− CAR-T cells. (A) The transient expression of CAR transgene 24 h after gene transfer (n = 3, 3 donors). (B) Number of CAR positive T cells at day 14 (n = 5, 3 donors). (C) Representative expression and phenotypes of CD19 CAR-T, and expression of exhaustion markers on CAR-T cells assessed by flow cytometry. The gating control (left, blue) for CAR expression showed CAR-T cells only, stained by anti-human IgG Fc fragment specific antibody conjugated to FITC, the gating of CAR expression (left, red) showed CAR-T cells combined with the recombinant human CD19 Fc chimera protein and secondary stained by anti-human IgG Fc fragment specific antibody conjugated to FITC. (D) The phenotype and exhaustion marker of CD19 CAR-T cells are represented (n = 3–6, 3 donors). (E) A volcano plot showing genes with adjusted FDR <0.05 that are differentially expressed in CD45RA+ CAR-T compared with CD45RA− CAR-T. (F) Reactome pathway analysis showed that several gene pathways were significantly downregulated in CD45RA+ CAR-T cells compared with CD45RA− CAR-T. Both red and blue circles showed downregulation of gene pathways in CD45RA+ CAR-T, and their colors represented adjusted p-values. (G) Transcriptome profiling about expression of T-cell activation genes (highlighted by red underline) and exhaustion genes (highlighted by blue underline) in CD45RA+ CAR-T and CD45RA− CAR-T, before antigen stimulation (left) and after antigen stimulation (right). Row min denotes lowest Z-score and row max denotes highest Z-score. All data are presented as means ± standard deviation. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001.