Figure 3 from Autologous Thymic Organoids Support Functional T-cell Education and Enhance Antitumor Immunity in Humanized Mice with Melanoma Xenografts

Examination of T-cell development and activity in the mHMTA. A, Representative cytometry of the human CD45+ cells and HSPCs present in mHM and mHMTA bone marrow. B, Comparison of the average percentage of human CD45+ cells and CD34+ HSPCs within this CD45+ cell population within mHM and mHMTA bone marrow (P = 0.118 and 0.379, respectively, Wilcoxon rank-sum test). C, Comparison of peripheral human CD45+ lymphocytes in mHM and mHMTA (P = 0.728, Wilcoxon rank-sum test) and of CD19+ B cells in mHM and mHMTA (0.224, Wilcoxon rank-sum test). D, Cytometric analysis and a comparison of human CD45+ cells, as well as T-cell and B-cell populations, found in the spleens of mHM and mHMTA. E, Comparison of the average human CD45+ cell populations, human T cells, and human B cells on mHM and mHMTA spleens (P = 0.844, 0.450, and 0.646, Wilcoxon rank-sum test, respectively). F, IFNγ expression was detected in the spleen of both mHM and mHMTA (P < 0.005). G, Gating and representative cytometry of T cells in mHM and mHMTA show relative populations used to analyze T-cell activation. Purple boxes in the first panels show the T-cell populations gated in the remaining scatter plots. The red boxes show the CD8+ T-cell populations gated for subsequent analysis. H, Representative cytometry of the CD8+ T cells, gated from the red boxes in the previous panel. I, Comparisons of the ratio of HLA-DR + PD1– T cells with all HLA-DR+ T cells shows an increasingly activated T-cell population in mHMTA (P = 0.0207, two-group t test) and a nonsignificantly increased population of CD4+ T cells. J, Comparison of the HLA-DR + PD1– population with the total HLA-DR+ T-cell population among CD8+ T cells also reveals a trend toward increased activation in mHMTA. P values: *, ≤0.05.