CARFH80 construct and leukemic CD19/CD22 expression in patie

determined by flow cytometry with a population of CD22-negative non-B cells with similar cell size in the same staining tube (blue) as a negative control for evaluating CD22 expression level. The blasts were defined based on the combined analysis of multiple markers. The number represents the proportion of blasts (red) in the four quadrants. h Histogram and mean fluorescence intensity of CD22 cell-surface expression in patients 03 and 04. ， unpaired two-sample t-test was used for statistical analysis. The brown， determined by flow cytometry. c Nuclear factor of activated T cells (NFAT) reporter assay in T cells transduced with different CD22-BBz variants。

after co-culturing with CD22high Raji， and CD22- K562 cells， 0.5:1。

a Schematic of the recombinant lentiviral vectors encoding the full-human CD22-BBz variants. The expression of CAR transgene is under the control of the elongation factor 1α promoter. CD8 (H/TM)。

CD22-BBz 27， JVM-2，。

CD22low JVM-2。

or cultured in medium alone in triplicates. d Bar chart showing CD107a expression in specific CD22-BBz T cell clones after co-culturing with CD22high Raji， Nalm6， Thoseaasigna virus 2A. b Histogram of CD22 expression on Raji， and K562 cells， ****P  0.0001. f Membrane proteome array for CD22-BBz. g Dot plots showing the proportions of blasts (red) and non-tumor cells (black) among all mononuclear cells from patients at the indicated time points， and contour plots showing CD22 and CD19 expression on blasts (red) before CD22-CARYK002 and CD22-CARFH80 T cell therapy， 1:1， Jurkat， green， respectively. *P  0.05。

Reh cells， and 0:1. % specific lysis = (spontaneous relative light unit (RLU) − test RLU)/(spontaneous death RLU) × 100. A two-tailed， CD8α hinge and transmembrane domains; EGFR， epidermal growth factor receptor; T2A， and U266 cells， CD22− K562， as determined by flow cytometry. CD107a degranulation rates were calculated as the percentages of CD107a+ cells among CAR+CD8+ T cells and indicated above each bar. e Cytolytic effects of specific CD22-BBz T cell clones against Nalm6 and Reh cells during co-culturing in triplicates at E:T ratios of 2:1， or control T cells， **P  0.01， and blue asterisks indicate the comparison of the cytolytic effects of CD22-BBz 80 with those of CD22-BBz 36。

CD22low JVM-2， ***P  0.001。