Fig. 6
The PE8T cells reduced exhaustion and increased oxygen consumption rate inculcated with CD19 + PDL1 + tumor cells. A. Proliferation curves of different CAR-T cells from day 3 to day 11 after isolation (mean ± SD, n = 3). Results are representative of triplicate with T cells from one healthy donor. B. The stage of T cell differentiation and development is defined as: naïve T (TN): CD45RA + CD45RO−, stem cell memory T (TSCM): CD45RA + CD45RO+, central memory T (TCM): CD45RA + CCR7+, and effect memory T (TEM): CD45RA − CCR7−. Phenotypic detection was performed on day 8 after isolation (mean ± SD, n = 3). Results are representative of triplicate with T cells from one healthy donor. 2G CAR-T vs.PE8T CAR-T.C. Representative GSEA results from running the 2G CAR-T vs. PE8T CAR-T cells rank list about differentiation gene ontology sets (mean ± SD, n = 3). Results are representative of triplicate with T cells from one healthy donor. CAR-T cells were collected after stimulation with NALM-6-PDL1 cells at E:T of 1:1 for 48 h. D. The OCR, basal OCR and maximum OCR of 2G CAR-T and PE8T CAR-T cells in culture under basal metabolic conditions and in response to mitochondrial inhibitors, as specified in the experimental procedures. E. ECAR, basal ECAR and maximum ECAR of 2G CAR-T and PE8T CAR-T cells in culture under basal metabolic conditions and in response to mitochondrial inhibitors, as specified in the experimental procedures. Data represent at least triplicate performed with cells from one healthy human donor plotted as mean ± SD (*P < 0.05; **P < 0.01; ***P < 0.005.). One of three independent experiments is presented (n = 3). Data are represented as mean ± SD.F. Representative GSEA results from running the 2G CAR-T vs. PE8T CAR-T cells rank list about metabolism gene ontology sets (mean ± SD, n = 3). Results are representative of triplicate with T cells from one healthy donor. CAR-T cells were collected after stimulation with NALM-6-PDL1 cells at E:T of 1:1 for 48 h. One of three independent experiments is presented (n = 3). Data are represented as mean ± SD. ECAR, extracellular acidification rate; GSEA, gene set enrichment analysis; OCR, oxygen consumption rate