Fig. 3: Increased cholesterol efflux from M2 macrophages in patients with PD.

a Percentages of M2 macrophages in patients with PD before CAR-T cell therapy, during CAR-T cell expansion, and during disease progression. b Gene Set Enrichment Analysis for M2 macrophages of patients with PD at different timepoints. c In vitro, the culture supernatants collected from DB cells were used to culture control cells, M2 cells, and ABCA1-knockdown M2 (M2^shABCA1) cells. d Cytomembrane cholesterol levels in M2 macrophages were assessed via immunofluorescence using the Vybrant Alexa Fluor 488 lipid raft labeling kit. e Coculture models were constructed to assess the effects of M2 macrophages on CAR-T cell cytotoxicity against CD19-expressing DLBCL cells (DB cells) and the impact of CAR19 cells on M2 macrophages’ total cholesterol levels either within cells or secreted into the coculture medium. f Total cholesterol levels in the cocultured M2 macrophages and cell-free cocultured media were measured in the above indicated groups. g Cytotoxic effect of CAR19 cells on CD19-expressing DLBCL cells (DB) and CD19-nonexpressing acute promyelocytic leukemia cells (HL60) was detected using a luciferase-based CTL assay. Data are shown as mean ± s.e.m. Statistical analysis was performed using two-way ANOVA with Tukey’s multiple comparison tests. n = 3 for before CAR-T cell therapy group, n = 3 for at CAR-T cell expansion group, and n = 2 for at disease progression group in patients with PD.