Fig. 4

Immunophenotypic characterization of a long-lived CAR T population in a single patient. (A) Heatmap shows normalized expression of 33 surface markers of CAR T cells for nine clusters identified with FlowSom. CAR T population was gated upon CD45⁺CD11b⁻CD11c⁻CD66b⁻CD19⁻CD14⁻CD7⁺CD3⁺VHH⁺. Relative frequencies are displayed as a bar graph to the right bottom. (B) t-SNE map displays CAR T cells in the four samples of different time points. Cells are colored by FlowSom cluster. (C) The left upper panel shows three subsets of CD4⁺, CD8⁺ and DN CAR T cells in the four samples of different time points. The left bottom panel shows the cluster disposition of each CAR T cell subset. The right panel shows the frequencies of 9 clusters in individual sample. DN: CD4 and CD8 double-negative. (D) The violin plots display the immunophenotypic expression of Ki67, HLA-DR, CCR7, GZMB and PD-1 of CD4⁺, CD8⁺ and DN CAR T cells of the four samples. p values were statistically calculated by comparing the mean marker expression of the sample of 5 years to that of 2 months. (E) The violin plots compare the expression levels of Ki67, GZMB, CD107a, PD-1 and CD57 between the CAR T cells (CD3⁺VHH⁺) and the host T cells (CD3⁺VHH⁻). The abbreviations for month and year are mo and yr, respectively