PMC7292833_01
Tumor B7-H3 expression and characteristics of chimeric antigen receptor (CAR)-T cells. (a) Immunohistochemical analysis of paraffin-embedded tumor tissues collected before CAR-T therapy. (b) Immunofluorescent staining (red) of B7-H3 in anaplastic meningioma primary cells. (c) CAR component construction in the lentivirus vector. (d) Flow cytometric analysis of surface CAR expression and the T-cell markers CD25, CD69, CD45RO, CD62L, programmed cell death-1, and T-cell immunoglobulin and mucin domain-containing protein 3. CD19 was used for CAR detection. (e) Images of CAR-T cells cocultured with autologous tumor (anaplastic meningioma) cells at E/T ratios of 5:1 and 10:1. CD19-targeted CAR-T cells (CD19 CAR) served as controls. (f)
51Cr release assays of CAR-T cells against primary tumor cells at different E/T ratios. (g) Cytokine (interferon gamma and IL2) secretion levels were measured to analyse immunological changes in B7-H3/CD19-targeted CAR-T cells cocultured with primary tumor cells (40,000 T cells to 10,000 tumor cells) for 12 h using enzyme-linked immunosorbent assays. *P < 0.05. One representative image from two experiments is shown in b and e. For f and g, the results show cumulative data from two independent experiments.
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A 49-year-old woman presented with multiple recurrent anaplastic meningioma (WHO grade III, Ki67 expression: 30%) accompanied by disorder of the right limb (Supplementary figure 1b). As shown in Figure 1a, immunohistochemistry (IHC) indicated high and homogeneous B7-H3 expression in tumor samples, with a histochemistry score of 180. Consistent with the IHC result, immunofluorescence imaging of tumor-isolated primary cells also confirmed the overexpression of B7-H3 (Figure 1b).