Fig. 3: Crosstalk between CAR T cell and leukaemia bone marrow niche.
a, Representative images showing ICAM-1 (green) expression on vascular cells (CD31+ in orange) from leukaemia chips treated with, respectively, Mock T cell (top left) and CAR T cell (bottom left) for 2 days. Representative images were from one of the three technical replicates with similar results (n = 3). Quantification data of ICAM-1 and CD31 intensity, corresponding to the white dash lines in the images. b, Representative images showing HLA-DR expression on monocytic cells (CD14+) from leukaemia chips respectively treated with Mock T cell and CAR T cell for 2 days. Representative images and data were collected from four independent experiments (n = 4). Unpaired, two-sided, Student’s t-test, mean and s.e.m. c, Relative cytokine secretion from leukaemia chips established with CD19+ or CD19− leukaemia blasts, which were treated with 10,000 of CAR T cells (CAR + CD19+ and CAR + CD19−), Mock T cells (Mock + CD19+ and Mock + CD19−) or left untreated (CD19+ and CD19−). The results showed that CAR + CD19+ demonstrated excessive on-chip production of cytokines, such as immune stimulatory cytokines (for example, MIP-1α), chemokines (for example, MIP-1β and RANTES) and inflammatory cytokines (for example, MCP-2), among many others (Supplementary Fig. 6c). d, MIP-1β cytokine secretion from leukaemia chips established with (immunocompetent) or without (immunocompromised) bone marrow mononuclear cells, where MIP-1β secretion was enhanced in immunocompetent leukaemia chips. Cytokine secretion profiles in c and d were examined from chips on day 2 by using a Human Inflammation Array C3 membrane kit, and data were collected from three independent experiments (n = 3). e, ELISA quantification of MIP-1β secretion on day 2 from leukaemia immunocompetent and immunocompromised chips treated with 10,000 of CAR T cells. Data were collected from three independent experiments (n = 3). Two-way ANOVA, mean and s.e.m. f, Analysis of DEGs of vascular cells from leukaemia chips treated with CAR T cell (HD CAR+) or Mock T cell (Mock+) for 2 days. Two-sided Wilcoxon rank sum test. g, DEGs of macrophages from leukaemia chips treated with CAR T cell (HD CAR+) or Mock T cell (Mock+) for 2 days. Two-sided Wilcoxon rank sum test.
